600 results on '"Medrano, Juan F"'
Search Results
152. A 500-kb YAC and BAC Contig Encompassing the High-Growth Deletion in Mouse Chromosome 10 and Identification of the MurineRaidd/CraddGene in the Candidate Region
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Horvat, Simon, primary and Medrano, Juan F., additional
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- 1998
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- View/download PDF
153. Effects of Milk Protein Genotypes on the Variation for Milk Production Traits of Holstein and Jersey Cows in California
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Ojala, Matti, primary, Famula, Thomas R., additional, and Medrano, Juan F., additional
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- 1997
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154. Polymorphisms of Bovine β-Lactoglobulin Promoter and Differences in the Binding Affinity of Activator Protein-2 Transcription Factor
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Lum, Lorraine S., primary, Dovč, Peter, additional, and Medrano, Juan F., additional
- Published
- 1997
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155. Rapid verification of meiotic gynogenesis and polyploidy in white sturgeon (Acipenser transmontanus Richardson)
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Van Eenennaam, Alison L., primary, Van Eenennaam, Joel P., additional, Medrano, Juan F., additional, and Doroshov, Serge I., additional
- Published
- 1996
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156. Thehigh growth(hg) Locus Maps to a Deletion in Mouse Chromosome 10
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Horvat, Simon, primary and Medrano, Juan F., additional
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- 1996
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157. Gender and Single Nucleotide Polymorphisms in MTHFR, BHMT, SPTLC1, CRBP2, CETP, and SCARB1 Are Significant Predictors of Plasma Homocysteine Normalized by RBC Folate in Healthy Adults.
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Clifford, Andrew J., Kehui Chen, McWade, Laura, Rincon, Gonzalo, Seung-Hyun Kim, Holstege, Dirk M., Owens, Janel E., Bitao Liu, Müller, Hans-Georg, Medrano, Juan F., Fadel, James G., Moshfegh, Alanna J., Baer, David J., and Novotny, Janet A.
- Subjects
SINGLE nucleotide polymorphisms ,HOMOCYSTEINE in the body ,ERYTHROCYTES ,REGRESSION analysis ,BODY mass index ,BLOOD testing ,GENETIC regulation ,PREDICTION theory - Abstract
Using linear regression models, we studied the main and 2-way interaction effects of the predictor variables gender, age, BMI, and 64 folateA/itamin B-12/homocysteine (Hcyl/lipid/cholesterol-related single nucleotide polymorphisms (SNP) on log-transformed plasma Hey normalized by RBC folate measurements (nHcy) in 373 healthy Caucasian adults (50% women). Variable selection was conducted by stepwise Akaike information criterion or least angle regression and both methods led to the same final model. Significant predictors (where Pvalues were adjusted for false discovery rate) included type of blood sample [whole blood (WB) vs. plasma-depleted WB; P < 0.001 ] used for folate analysis, gender (P< 0.001), and SNP in genes SPTLC1 (rs11790991 ; P= 0.040), CRBP2 (rs2118981 ; P< 0.001 ), BHMT (rs3733890; P = 0.019), and CETP (rs5882; P = 0.017). Significant 2-way interaction effects included gender x MTHFR (rs1801131 ; P= 0.012), gender x CRBP2 (rs2118981; P= 0.011), and gender X SCARB1 (rs83882; P= 0.003). The relation of nHcy concentrations with the significant SNP (SPTLC1, BHMT, CETP, CRBP2, MTHFR, and SCARB1) is of interest, especially because we surveyed the main and interaction effects in healthy adults, but it is an important area for future study. As discussed, understanding Hey and genetic regulation is important, because Hey may be related to inflammation, obesity, cardiovascular disease, and diabetes mellitus. We conclude that gender and SNP significantly affect nHcy. [ABSTRACT FROM AUTHOR]
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- 2012
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158. Identification of Structural Variants Associated with Mastitis in Holstein Dairy Cows Using Whole Genome Sequencing and RNA-sequencing.
- Author
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Asselstine, Victoria, Medrano, Juan F. F., Muniz, Malane M. M., and Cánovas, Angela
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WHOLE genome sequencing , *LINCRNA , *DAIRY cattle , *RNA sequencing , *MASTITIS , *GENETIC variation , *EXOMES - Abstract
For the dairy industry, animal welfare and economic viability are key factors in determining the long-term sustainability of the industry. One challenge in lactating dairy cows is mastitis, as there are significant costs associated with each mastitis case due to treatment, milk loss, and potential cow culling. Differentially expressed (DE) genes, DE mRNA isoforms and DE long non-coding RNA (lncRNA) candidates were previously identified by our group between healthy and mastitic samples from six Holstein dairy cows. In total, 7 candidate genes, 5 mRNA isoforms and 4 lncRNA were targeted as they play an important role due to their association with mastitis or the immune system. The aim of the current research is to identify new structural variants in the transcriptome of these previously identified potential candidates using RNASequencing. These structural variants could occur in both coding and intergenic regions of the genome and potentially impact the amino acid that is transcribed, creating a new functional or non-functional protein. In addition, whole genome sequencing (WGS) analysis was performed using hair samples collected from ten Holstein dairy cows (first lactation). Five of these animals had no previous reports of mastitis and five animals had at least one report of mastitis in her lifespan. The WGS results will be used to identify additional structural variants, especially within introns, of the previously identified candidates that could cause a genetic variation in individuals and an association to their susceptibility to mastitis disease. [ABSTRACT FROM AUTHOR]
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- 2021
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159. Lack of Socs2Expression Causes the High-Growth Phenotype in Mice
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Horvat, Simon and Medrano, Juan F.
- Abstract
Characterizing causal molecular defects in mouse models of overgrowth or dwarfism helps to identify the key genes and pathways that regulate the growth process. We report here the molecular basis for high growth (hg), a spontaneous mutation that causes a 30–50% increase in postnatal growth. We conclude that hgis an allele of the suppressor of cytokine signaling 2 (Socs2), a member of a family of regulators of cytokine signal transduction. We demonstrate mapping of Socs2to the hgregion, lack of Socs2mRNA expression, a disruption of the Socs2locus in high-growth (HG) mice, and a similarity of phenotypes of HG mice and Socs2−/−mice generated by gene targeting. Characteristics of the HG phenotype suggest that Socs2deficiency affects growth prenatally and postnatally most likely through deregulating the growth hormone (GH)/insulin-like growth factor I (IGF1). These results demonstrate a critical role for Socs2in controlling growth.
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- 2001
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160. Infertility in a Line of Mice with the high growthMutation Is Due to Luteal Insufficiency Resulting from Disruption at the Hypothalamic-Pituitary Axis1
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Cargill, Shelley L., Medrano, Juan F., and Anderson, Gary B.
- Abstract
Animals with extreme body growth frequently experience poor reproductive performance, but the cause for this association has not been clearly established. A line of mice homozygous for the high growth(hg) mutation, a mutation that has a major effect on post-weaning growth, exhibits several reproductive deficits including an abnormally high incidence of luteal failure. The objective of the present study was to investigate luteal failure in high-growth (HG) mice during pregnancy and to determine whether the cause of the apparent luteal failure resides in the ovary or the hypothalamic-pituitary unit. In HG females with a demonstrated history of infertility, progesterone injections (1 mg s.c. daily) beginning on Day 1 postcoitum (p.c.) increased the proportion of animals pregnant at Day 17 of gestation. Twice-daily injections of 100 μg of ovine prolactin (PRL) in alkaline saline given to another group of females beginning on Day 1 p.c. increased the proportion of HG females that were pregnant on Day 6 of gestation compared with saline-injected HG females, although PRL did not increase the pregnancy rate in HG females when compared with a group of noninjected control females. When ovaries from HG females were transplanted into ovariectomized congenic C57 hosts, the C57 graft hosts displayed normal estrous cycles, and upon mating the majority of graft hosts became pregnant. In contrast, when ovaries from C57 females were transplanted into ovariectomized HG hosts, the HG graft hosts displayed normal estrous cycles, but upon mating most were unable to maintain pregnancy. These results suggest that the hypothalamic-pituitary unit of the HG female provides an inadequate signal to the ovaries to maintain pregnancy. Luteal failure in HG females may be due to insufficient PRL as required for establishment and early maintenance of the CL during pregnancy in mice.
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- 1999
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161. Differential expression of bovine β-lactoglobulin A and B promoter variants in transiently transfected HC11 cells
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*, JOSEP M. FOLCH, , , *, PETER DOV *, §, and MEDRANO, JUAN F.
- Abstract
Quantification of β-lactoglobulin A and B in the milk of heterozygous animals has revealed a differential content of these two variants. Nucleotide sequence analysis of the first 733 bp of the bovine β-lactoglobulin promoter has indicated the presence of ten polymorphic sites, nine of them being allele A and B specific mutations. To study the differential expression of these alleles, constructs containing 753 bp long fragments of the bovine β-lactoglobulin A and B associated promoters were used in transient transfection experiments in HC11 cells. A differential transcription activity directed by the A and B specific promoters was consistently observed. The relative expression levels were 57% for β-lactoglobulin A and 43% for β-lactoglobulin B promoters. An allele-specific mutation has been reported to have a differential binding affinity to the activator protein-2 between the β-lactoglobulin A and B promoters. However, experiments in HC11 cells where the activator protein-2 binding sites were mutated in both β-lactoglobulin A and B promoters showed no major differences in activity between the mutated and native promoters.
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- 1999
162. A comparative analysis of chromatin accessibility in cattle, pig, and mouse tissues.
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Halstead, Michelle M., Kern, Colin, Saelao, Perot, Wang, Ying, Chanthavixay, Ganrea, Medrano, Juan F., Van Eenennaam, Alison L., Korf, Ian, Tuggle, Christopher K., Ernst, Catherine W., Zhou, Huaijun, and Ross, Pablo J.
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SWINE ,CATTLE ,TRANSCRIPTION factors ,UNGULATES ,COMPARATIVE studies ,MICE - Abstract
Background: Although considerable progress has been made towards annotating the noncoding portion of the human and mouse genomes, regulatory elements in other species, such as livestock, remain poorly characterized. This lack of functional annotation poses a substantial roadblock to agricultural research and diminishes the value of these species as model organisms. As active regulatory elements are typically characterized by chromatin accessibility, we implemented the Assay for Transposase Accessible Chromatin (ATAC-seq) to annotate and characterize regulatory elements in pigs and cattle, given a set of eight adult tissues. Results: Overall, 306,304 and 273,594 active regulatory elements were identified in pig and cattle, respectively. 71,478 porcine and 47,454 bovine regulatory elements were highly tissue-specific and were correspondingly enriched for binding motifs of known tissue-specific transcription factors. However, in every tissue the most prevalent accessible motif corresponded to the insulator CTCF, suggesting pervasive involvement in 3-D chromatin organization. Taking advantage of a similar dataset in mouse, open chromatin in pig, cattle, and mice were compared, revealing that the conservation of regulatory elements, in terms of sequence identity and accessibility, was consistent with evolutionary distance; whereas pig and cattle shared about 20% of accessible sites, mice and ungulates only had about 10% of accessible sites in common. Furthermore, conservation of accessibility was more prevalent at promoters than at intergenic regions. Conclusions: The lack of conserved accessibility at distal elements is consistent with rapid evolution of enhancers, and further emphasizes the need to annotate regulatory elements in individual species, rather than inferring elements based on homology. This atlas of chromatin accessibility in cattle and pig constitutes a substantial step towards annotating livestock genomes and dissecting the regulatory link between genome and phenome. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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163. Direct and Correlated Responses to Selection for Body Size and Oxygen Consumption in Tribolium
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Medrano, Juan F. and Gall, G. A. E.
- Abstract
Between family selection was applied to four lines of Tribolium castaneum originating from the same base population to evaluate direct and correlated responses in body size and metabolic activity. Each line consisted of 24 full-sib families, and selection was performed at an intensity of 25% for six generations. Selection criteria were: Line 1 - low oxygen consumption per unit body weight at 12 days of age (02/mg), Line 2 — high 12-day larva weight, Line 3 — high 18-day pupa weight, and Line 4 — randomly selected.Significant direct responses were observed in all three lines; realized heritabilities were .44 ± .07, .20 ± .04 and .36 ± .04 for 02/mg, larva weight and pupa weight, respectively. Significant correlated responses were observed between larva and pupa weight, and between pupa weight and 02/mg. Asymmetry in the realized genetic correlations, particularly for pupa weight and 02/mg, indicated that different genetic systems responded to selection when selection criteria were interchanged.The growth curve response of the lines at generation 6 indicated that the developmental time had increased in lines 1 and 3 as compared to line 4, whereas line 2 realized no major developmental change as a consequence of selection.The cessation of response for low 02/mg in the early generations of selection was attributed to a probable approach to a physiological limit in metabolic activity. Causes of asymmetry in correlated responses between pupa weight and 02/mg are discussed in terms of the biological efficiency of the organisms. Inferences are made concerning further applications of the present findings to selection studies of growth in warm blooded organisms in which the objective is to obtain larger but metabolically more efficient animals.
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- 1976
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164. Non-Coding RNA Sequencing of Equine Endometrium During Maternal Recognition of Pregnancy.
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Klohonatz, Kristin M., Coleman, Stephen J., Cameron, Ashley D., Hess, Ann M., Reed, Kailee J., Canovas, Angela, Medrano, Juan F., Islas-Trejo, Alma D., Kalbfleisch, Ted, Bouma, Gerrit J., and Bruemmer, Jason E.
- Subjects
NUCLEOTIDE sequence ,NON-coding RNA ,ENDOMETRIUM ,PREGNANCY ,NUCLEOTIDES - Abstract
Maternal recognition of pregnancy (MRP) in the mare is not well defined. In a non-pregnant mare, prostaglandin F
2α (PGF) is released on day 14 post-ovulation (PO) to cause luteal regression, resulting in loss of progesterone production. Equine MRP occurs prior to day 14 to halt PGF production. Studies have failed to identify a gene candidate for MRP, so attention has turned to small, non-coding RNAs. The objective of this study was to evaluate small RNA (<200 nucleotides) content in endometrium during MRP. Mares were used in a cross-over design with each having a pregnant and non-mated cycle. Each mare was randomly assigned to collection day 11 or 13 PO (n = 3/day) and endometrial biopsies were obtained. Total RNA was isolated and sequencing libraries were prepared using a small RNA library preparation kit and sequenced on a HiSeq 2000. EquCab3 was used as the reference genome and DESeq2 was used for statistical analysis. On day 11, 419 ncRNAs, representing miRNA, snRNA, snoRNA, scaRNA, and vaultRNA, were different between pregnancy statuses, but none on day 13. Equine endometrial ncRNAs with unknown structure and function were also identified. This study is the first to describe ncRNA transcriptome in equine endometrium. Identifying targets of these ncRNAs could lead to determining MRP. [ABSTRACT FROM AUTHOR]- Published
- 2019
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165. Coding RNA Sequencing of Equine Endometrium during Maternal Recognition of Pregnancy.
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Klohonatz, Kristin M., Coleman, Stephen J., Islas-Trejo, Alma D., Medrano, Juan F., Hess, Ann M., Kalbfleisch, Ted, Thomas, Milton G., Bouma, Gerrit J., and Bruemmer, Jason E.
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NUCLEOTIDE sequence ,ENDOMETRIUM ,CORPUS luteum ,PREGNANCY ,GENETIC engineering - Abstract
Equine maternal recognition of pregnancy (MRP) is a process whose signal remains unknown. During MRP the conceptus and endometrium communicate to attenuate prostaglandin F
2α (PGF) secretion, sparing the corpus luteum and maintaining progesterone production. Recognition of a mobile conceptus by the endometrium is critical by days 14–16 post-ovulation (PO), when endometrium produces PGF, initiating luteolysis. The objective of this study was to evaluate endometrial gene expression changes based upon pregnancy status via RNA sequencing. This experiment utilized a cross-over design with each mare serving as both a pregnant and non-mated control on days nine, 11, and 13 PO (n = 3/status/day). Mares were randomly assigned to collection day and pregnancy confirmed by terminal uterine lavage at the time of endometrial biopsy. Total RNA was isolated and libraries prepared using Illumina TruSeq RNA sample preparation kit. Reads were mapped and annotated using HISAT2 and Stringtie. Expression values were evaluated with DESEQ2 (P ≤ 0.05 indicated significance). On day nine, 11, and 13 there were 1435, 1435 and 916 significant transcripts, respectively. Multiple genes with splice variants had different expression patterns within the same day. These are the first data to evaluate the endometrial transcriptome during MRP on days nine, 11, and 13. [ABSTRACT FROM AUTHOR]- Published
- 2019
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166. Protein requirement of hatchery-produced juvenile white sturgeon (Acipenser transmontanus
- Author
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Moore, Brendan J., primary, Hung, Silas S.O., additional, and Medrano, Juan F., additional
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- 1988
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167. GROWTH RATE, BODY COMPOSITION, CELLULAR GROWTH AND ENZYME ACTIVITIES IN LINES OF TRIBOLIUM CASTANEUM SELECTED FOR 21-DAY PUPA WEIGHT
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Medrano, Juan F, primary and Gall, G A E, additional
- Published
- 1976
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168. FOOD CONSUMPTION, FEED EFFICIENCY, METABOLIC RATE AND UTILIZATION OF GLUCOSE IN LINES OF TRIBOLIUM CASTANEUM SELECTED FOR 21-DAY PUPA WEIGHT
- Author
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Medrano, Juan F, primary and Gall, G A E, additional
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- 1976
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169. Identification of one polymorphism from the PAPP-A2 gene associated to fertility in Romosinuano beef heifers raised under a subtropical environment.
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Luna-Nevárez, Pablo, Rincón, Gonzalo, Medrano, Juan F., Riley, David G., Chase Jr., Chad C., Coleman, Sam W., DeAtley, Kasey L., Islas-Trejo, A., Silver, Gail A., and Thomas, Milton G.
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SINGLE nucleotide polymorphisms , *CATTLE breeding , *CATTLE fertility , *HEIFERS , *ALLELES , *CATTLE genetics , *CATTLE reproduction - Abstract
The objective was to identify single nucleotide polymorphisms (SNP) associated to fertility in cows raised under a subtropical environment. Re-sequencing of nine genes associated to GH-IGF endocrine pathway, which are located in bovine chromosomes 5, 16 and 20, identified 73 SNP useful for associative genetic studies; however, only seven resulted as polymorphic and unique to the Romosinuano breed. Then, DNA samples were extracted from 129 beef heifers and used to determine genotypes corresponding to each SNP. Mixed model analysis identified one SNP from the PAPP-A2 gene (C/T, rs110490898) as predictor (P<0.05) of reproductive performance. The allele T was the most favorable allele (P<0.05), because it was associated with lower age at first calving (-37.1 ± 14.4 d) and age at second calving (-65.43 ± 30.8 d). In the contrast analysis, the linear term resulted as significant and quadratic term as non significant, which suggested an additive effect of the alleles. These results provide evidence to support the PAPP-A2 gene as a candidate gene associated to reproductive performance in heifers and first-calf cows from the Romosinuano breed, raised under a subtropical environment. [ABSTRACT FROM AUTHOR]
- Published
- 2012
170. Identificación de un polimorfismo del gen PAPP-A2 asociado a la fertilidad en vaquillas Romosinuano criadas en subtrópico.
- Author
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Luna-Nevárez, Pablo, Rincón, Gonzalo, Medrano, Juan F., Riley, David G., Chase Jr., Chad C., Coleman, Sam W., DeAtley, Kasey L., Islas-Trejo, A., Silver, Gail A., and Thomas, Milton G.
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SINGLE nucleotide polymorphisms , *HEIFERS , *CATTLE breeding , *CATTLE fertility , *ALLELES , *DNA analysis - Abstract
El objetivo fue identificar polimorfismos de un solo nucleótido (SNP) asociados a la fertilidad en hembras bovinas criadas en subtropico. La re-secuenciación de nueve genes relacionados al eje endocrino GH-IGF, localizados en los cromosomas 5, 16 y 20 del bovino, identificó 73 SNP útiles para estudios genéticos asociativos; sin embargo, sólo siete resultaron polimórficos y exclusivos de la raza Romosinuano. Muestras de ADN se extrajeron de 129 vaquillas Romosinuano y usadas para determinar los genotipos correspondientes a cada SNP. Un análisis de modelos mixtos identificó únicamente a un polimorfismo del gen PAPP-A2 (C/T, rs110490898) como predictor (P<0.05) del comportamiento reproductivo. El alelo T fue el más favorable (P<0.05) ya que éste se asoció a una reducción tanto en la edad al primer parto (-37.1 ± 14.4 días), como en la edad al segundo parto (-65.43 ± 30.8 días). En el análisis de contrastes el término lineal resultó significativo (P<0.05), pero no el quadrático, lo cual sugiere un efecto aditivo de los alelos. Los resultados proporcionan evidencia para proponer al gen PAPP-A2, como candidato asociado al comportamiento reproductivo en vaquillas y vacas primerizas de la raza Romosinuano. [ABSTRACT FROM AUTHOR]
- Published
- 2012
171. Combining multi-OMICs information to identify key-regulator genes for pleiotropic effect on fertility and production traits in beef cattle.
- Author
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Fonseca, Pablo Augusto de Souza, Id-Lahoucine, Samir, Reverter, Antonio, Medrano, Juan F., Fortes, Marina S., Casellas, Joaquim, Miglior, Filippo, Brito, Luiz, Carvalho, Maria Raquel S., Schenkel, Flávio S., Nguyen, Loan T., Porto-Neto, Laercio R., Thomas, Milton G., and Cánovas, Angela
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BEEF cattle , *GENETIC regulation , *MEAT quality , *GENETIC pleiotropy , *DATA mining - Abstract
The identification of biological processes related to the regulation of complex traits is a difficult task. Commonly, complex traits are regulated through a multitude of genes contributing each to a small part of the total genetic variance. Additionally, some loci can simultaneously regulate several complex traits, a phenomenon defined as pleiotropy. The lack of understanding on the biological processes responsible for the regulation of these traits results in the decrease of selection efficiency and the selection of undesirable hitchhiking effects. The identification of pleiotropic key-regulator genes can assist in developing important tools for investigating biological processes underlying complex traits. A multi-breed and multi-OMICs approach was applied to study the pleiotropic effects of key-regulator genes using three independent beef cattle populations evaluated for fertility traits. A pleiotropic map for 32 traits related to growth, feed efficiency, carcass and meat quality, and reproduction was used to identify genes shared among the different populations and breeds in pleiotropic regions. Furthermore, data-mining analyses were performed using the Cattle QTL database (CattleQTLdb) to identify the QTL category annotated in the regions around the genes shared among breeds. This approach allowed the identification of a main gene network (composed of 38 genes) shared among breeds. This gene network was significantly associated with thyroid activity, among other biological processes, and displayed a high regulatory potential. In addition, it was possible to identify genes with pleiotropic effects related to crucial biological processes that regulate economically relevant traits associated with fertility, production and health, such as MYC, PPARG, GSK3B, TG and IYD genes. These genes will be further investigated to better understand the biological processes involved in the expression of complex traits and assist in the identification of functional variants associated with undesirable phenotypes, such as decreased fertility, poor feed efficiency and negative energetic balance. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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172. Genome-wide identification of tissue-specific long non-coding RNA in three farm animal species.
- Author
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Kern, Colin, Wang, Ying, Chitwood, James, Korf, Ian, Delany, Mary, Cheng, Hans, Medrano, Juan F., Van Eenennaam, Alison L., Ernst, Catherine, Ross, Pablo, and Zhou, Huaijun
- Subjects
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NON-coding RNA , *DOMESTIC animal genetics , *ANIMAL species , *ANIMAL classification , *GENOMES - Abstract
Background: Numerous long non-coding RNAs (lncRNAs) have been identified and their roles in gene regulation in humans, mice, and other model organisms studied; however, far less research has been focused on lncRNAs in farm animal species. While previous studies in chickens, cattle, and pigs identified lncRNAs in specific developmental stages or differentially expressed under specific conditions in a limited number of tissues, more comprehensive identification of lncRNAs in these species is needed. The goal of the FAANG Consortium (Functional Annotation of Animal Genomes) is to functionally annotate animal genomes, including the annotation of lncRNAs. As one of the FAANG pilot projects, lncRNAs were identified across eight tissues in two adult male biological replicates from chickens, cattle, and pigs. Results: Comprehensive lncRNA annotations for the chicken, cattle, and pig genomes were generated by utilizing RNA-seq from eight tissue types from two biological replicates per species at the adult developmental stage. A total of 9393 lncRNAs in chickens, 7235 lncRNAs in cattle, and 14,429 lncRNAs in pigs were identified. Including novel isoforms and lncRNAs from novel loci, 5288 novel lncRNAs were identified in chickens, 3732 in cattle, and 4870 in pigs. These transcripts match previously known patterns of lncRNAs, such as generally lower expression levels than mRNAs and higher tissue specificity. An analysis of lncRNA conservation across species identified a set of conserved lncRNAs with potential functions associated with chromatin structure and gene regulation. Tissue-specific lncRNAs were identified. Genes proximal to tissue-specific lncRNAs were enriched for GO terms associated with the tissue of origin, such as leukocyte activation in spleen. Conclusions: LncRNAs were identified in three important farm animal species using eight tissues from adult individuals. About half of the identified lncRNAs were not previously reported in the NCBI annotations for these species. While lncRNAs are less conserved than protein-coding genes, a set of positionally conserved lncRNAs were identified among chickens, cattle, and pigs with potential functions related to chromatin structure and gene regulation. Tissue-specific lncRNAs have potential regulatory functions on genes enriched for tissue-specific GO terms. Future work will include epigenetic data from ChIP-seq experiments to further refine these annotations. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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173. Functional Mapping of Quantitative Trait Loci That Interact With the hg Mutation to Regulate Growth Trajectories in Mice.
- Author
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Rongling Wu, Chang-Xing Ma, Wei Hou, Corva, Pablo, and Medrano, Juan F.
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GENETIC regulation , *LABORATORY mice , *ANIMAL genetics , *SEX chromosomes , *ALGORITHMS - Abstract
The high growth (hg) mutation increases body size in mice by 30-50%. Given the complexity of the genetic regulation of animal growth, it is likely that the effect of this major locus is mediated by other quantitative trait loci (QTL) with smaller effects within a web of gene interactions. In this article, we extend our functional mapping model to characterize modifier QTL that interact with the hg locus during ontogenetic growth. Our model is derived within the maximum-likelihood context, incorporated by mathematical aspects of growth laws and implemented with the EM algorithm. In an F2 population founded by a congenic high growth (HG) line and non-HG line, a highly additive effect due to the hg gene was detected on growth trajectories. Three QTL located on chromosomes 2 and X were identified to trigger significant additive and/or dominant effects on the process of growth. The most significant finding made from our model is that these QTL interact with the hg locus to affect the shapes of the growth process. Our model provides a powerful means for understanding the genetic architecture and regulation of growth rate and body size in mammals. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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174. Novel lncRNA regulatory elements in milk somatic cells of Holstein dairy cows associated with mastitis.
- Author
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Asselstine V, Medrano JF, Muniz MMM, Mallard BA, Karrow NA, and Cánovas A
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- Female, Cattle, Animals, Humans, Milk, Cell Count, Phenotype, RNA, Long Noncoding genetics, RNA, Long Noncoding metabolism, Mastitis metabolism
- Abstract
Despite regulatory elements such as long non - coding RNAs representing most of the transcriptome, the functional understanding of long non - coding RNAs in relation to major health conditions including bovine mastitis is limited. This study examined the milk somatic cell transcriptome from udder quarters of 6 Holstein dairy cows to identify differentially expressed long non - coding RNAs using RNA - Sequencing. Ninety - four differentially expressed long non - coding RNAs are identified, 5 of which are previously annotated for gene name and length, 11 are annotated for gene name and 78 are novel, having no gene name or length previously annotated. Significant inflammatory response and regulation of immune response pathways (false discovery rate < 0.05) are associated with the differentially expressed long non - coding RNAs. QTL annotation analysis revealed 31 QTL previously annotated in the genomic regions of the 94 differentially expressed long non - coding RNAs, and the majority are associated with milk traits. This research provides a better understanding of long non - coding RNAs regulatory elements in milk somatic cells, which may enhance current breeding strategies for more adaptable or high mastitis resistant cattle., (© 2024. The Author(s).)
- Published
- 2024
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175. Single-Step Genome Editing of Small Ruminant Embryos by Electroporation.
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Mahdi AK, Medrano JF, and Ross PJ
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- Animals, CRISPR-Cas Systems, Electroporation, Goats genetics, Ruminants, Sheep genetics, Zygote, RNA, Small Untranslated genetics, Gene Editing
- Abstract
We investigated the possibility of single-step genome editing in small ruminants by CRISPR-Cas9 zygote electroporation. We targeted SOCS2 and PDX1 in sheep embryos and OTX2 in goat embryos, utilizing a dual sgRNA approach. Gene editing efficiency was compared between microinjection and three different electroporation settings performed at four different times of embryo development. Electroporation of sheep zygotes 6 h after fertilization with settings that included short high-voltage (poring) and long low-voltage (transfer) pulses was efficient at producing SOCS2 knock-out blastocysts. The mutation rate after CRISPR/Cas9 electroporation was 95.6% ± 8%, including 95.4% ± 9% biallelic mutations; which compared favorably to 82.3% ± 8% and 25% ± 10%, respectively, when using microinjection. We also successfully disrupted the PDX1 gene in sheep and the OTX2 gene in goat embryos. The biallelic mutation rate was 81 ± 5% for PDX1 and 85% ± 6% for OTX2. In conclusion, using single-step CRISPR-Cas9 zygote electroporation, we successfully introduced biallelic deletions in the genome of small ruminant embryos.
- Published
- 2022
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176. Functional annotations of three domestic animal genomes provide vital resources for comparative and agricultural research.
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Kern C, Wang Y, Xu X, Pan Z, Halstead M, Chanthavixay G, Saelao P, Waters S, Xiang R, Chamberlain A, Korf I, Delany ME, Cheng HH, Medrano JF, Van Eenennaam AL, Tuggle CK, Ernst C, Flicek P, Quon G, Ross P, and Zhou H
- Subjects
- Amino Acid Motifs, Animals, Animals, Domestic genetics, Chromatin Immunoprecipitation Sequencing, Enhancer Elements, Genetic genetics, Epigenesis, Genetic, Epigenomics, Genome-Wide Association Study, Mice, Organ Specificity genetics, Phylogeny, Polymorphism, Single Nucleotide, Transcription Factors genetics, Cattle genetics, Chickens genetics, Gene Expression Regulation genetics, Genome genetics, Regulatory Sequences, Nucleic Acid genetics, Swine genetics, Transcription Factors metabolism
- Abstract
Gene regulatory elements are central drivers of phenotypic variation and thus of critical importance towards understanding the genetics of complex traits. The Functional Annotation of Animal Genomes consortium was formed to collaboratively annotate the functional elements in animal genomes, starting with domesticated animals. Here we present an expansive collection of datasets from eight diverse tissues in three important agricultural species: chicken (Gallus gallus), pig (Sus scrofa), and cattle (Bos taurus). Comparative analysis of these datasets and those from the human and mouse Encyclopedia of DNA Elements projects reveal that a core set of regulatory elements are functionally conserved independent of divergence between species, and that tissue-specific transcription factor occupancy at regulatory elements and their predicted target genes are also conserved. These datasets represent a unique opportunity for the emerging field of comparative epigenomics, as well as the agricultural research community, including species that are globally important food resources.
- Published
- 2021
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177. De novo assembly of the cattle reference genome with single-molecule sequencing.
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Rosen BD, Bickhart DM, Schnabel RD, Koren S, Elsik CG, Tseng E, Rowan TN, Low WY, Zimin A, Couldrey C, Hall R, Li W, Rhie A, Ghurye J, McKay SD, Thibaud-Nissen F, Hoffman J, Murdoch BM, Snelling WM, McDaneld TG, Hammond JA, Schwartz JC, Nandolo W, Hagen DE, Dreischer C, Schultheiss SJ, Schroeder SG, Phillippy AM, Cole JB, Van Tassell CP, Liu G, Smith TPL, and Medrano JF
- Subjects
- Animals, Breeding methods, Genomics methods, RNA-Seq methods, RNA-Seq standards, Reference Standards, Sequence Analysis, DNA methods, Sequence Analysis, DNA standards, Breeding standards, Cattle genetics, Genome, Genomics standards, Polymorphism, Genetic
- Abstract
Background: Major advances in selection progress for cattle have been made following the introduction of genomic tools over the past 10-12 years. These tools depend upon the Bos taurus reference genome (UMD3.1.1), which was created using now-outdated technologies and is hindered by a variety of deficiencies and inaccuracies., Results: We present the new reference genome for cattle, ARS-UCD1.2, based on the same animal as the original to facilitate transfer and interpretation of results obtained from the earlier version, but applying a combination of modern technologies in a de novo assembly to increase continuity, accuracy, and completeness. The assembly includes 2.7 Gb and is >250× more continuous than the original assembly, with contig N50 >25 Mb and L50 of 32. We also greatly expanded supporting RNA-based data for annotation that identifies 30,396 total genes (21,039 protein coding). The new reference assembly is accessible in annotated form for public use., Conclusions: We demonstrate that improved continuity of assembled sequence warrants the adoption of ARS-UCD1.2 as the new cattle reference genome and that increased assembly accuracy will benefit future research on this species., (© The Author(s) 2020. Published by Oxford University Press.)
- Published
- 2020
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178. Deducing signaling pathways from parallel actions of arsenite and antimonite in human epidermal keratinocytes.
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Phillips MA, Cánovas A, Rea MA, Islas-Trejo A, Medrano JF, Durbin-Johnson B, Rocke DM, and Rice RH
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- Adrenal Cortex Hormones metabolism, Colony-Forming Units Assay, Ephrins metabolism, Gene Expression Profiling, Humans, Keratinocytes drug effects, Oncostatin M pharmacology, RNA, Messenger genetics, RNA, Messenger metabolism, Transcription Factors metabolism, Antimony pharmacology, Arsenites pharmacology, Epidermis metabolism, Keratinocytes metabolism, Signal Transduction drug effects, Signal Transduction genetics
- Abstract
Inorganic arsenic oxides have been identified as carcinogens in several human tissues, including epidermis. Due to the chemical similarity between trivalent inorganic arsenic (arsenite) and antimony (antimonite), we hypothesized that common intracellular targets lead to similarities in cellular responses. Indeed, transcriptional and proteomic profiling revealed remarkable similarities in differentially expressed genes and proteins resulting from exposure of cultured human epidermal keratinocytes to arsenite and antimonite in contrast to comparisons of arsenite with other metal compounds. These data were analyzed to predict upstream regulators and affected signaling pathways following arsenite and antimonite treatments. A majority of the top findings in each category were identical after treatment with either compound. Inspection of the predicted upstream regulators led to previously unsuspected roles for oncostatin M, corticosteroids and ephrins in mediating cellular response. The influence of these predicted mediators was then experimentally verified. Together with predictions of transcription factor effects more generally, the analysis has led to model signaling networks largely accounting for arsenite and antimonite action. The striking parallels between responses to arsenite and antimonite indicate the skin carcinogenic risk of exposure to antimonite merits close scrutiny.
- Published
- 2020
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179. Transcriptomic Profiles of Monocyte-Derived Macrophages in Response to Escherichia coli is Associated with the Host Genetics.
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Emam M, Cánovas A, Islas-Trejo AD, Fonseca PAS, Medrano JF, and Mallard B
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- Animals, Cattle, Macrophage Activation, Macrophages cytology, Macrophages immunology, Nitric Oxide metabolism, Nitric Oxide Synthase Type II metabolism, Phagocytosis, Phenotype, Polymorphism, Genetic, RNA, Messenger metabolism, Time Factors, Transcription Factors genetics, Transcription Factors metabolism, Escherichia coli pathogenicity, Genomics methods, Macrophages metabolism, Transcriptome
- Abstract
Reactive Nitrogen Species (RNS) are a group of bactericidal molecules produced by macrophages in response to pathogens in a process called oxidative burst. Nitric oxide (NO
- ) is a member of RNS produced from arginine by inducible Nitric Oxide Synthase (iNOS) enzyme. The activity of iNOS and production of NO- by macrophages following stimulation is one of the indicators of macrophage polarization towards M1/proinflammatory. Production of NO- by bovine monocyte-derived macrophage (MDM) and mouse peritoneal macrophages has been shown to be strongly associated with host genetic with the heritability of 0.776 in bovine MDM and 0.8 in mouse peritoneal macrophages. However, the mechanism of genetic regulation of macrophage response has remained less explored. In the current study, the transcriptome of bovine MDMs was compared between two extreme phenotypes that had been classified as high and low responder based on NO- production. The results showed that 179 and 392 genes were differentially expressed (DE) between high and low responder groups at 3 and 18 hours after exposure to Escherichia coli, respectively. A set of 11 Transcription Factors (TFs) (STAT1, IRF7, SPI1, STAT4, IRF1, HIF1A, FOXO3, REL, NFAT5, HIC1, and IRF4) at 3 hours and a set of 13 TFs (STAT1, IRF1, HIF1A, STAT4, ATF4, TP63, EGR1, CDKN2A, RBL1, E2F1, PRDM1, GATA3, and IRF4) at 18 hours after exposure to E. coli were identified to be differentially regulated between the high and low responder phenotypes. These TFs were found to be divided into two clusters of inflammatory- and hypoxia-related TFs. Functional analysis revealed that some key canonical pathways such as phagocytosis, chemotaxis, antigen presentation, and cell-to-cell signalling are enriched among the over-expressed genes by high responder phenotype. Based on the results of this study, it was inferred that the functional characteristics of bovine MDMs are associated with NO-based classification. Since NO- production is strongly associated with host genetics, this study for the first time shows the distinct proinflammatory profiles of macrophages are controlled by the natural genetic polymorphism in an outbred population. In addition, the results suggest that genetics can be considered as a new dimension in the current model of macrophage polarization which is currently described by the combination of stimulants, only.- Published
- 2020
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180. STAT6 , PBX2 , and PBRM1 Emerge as Predicted Regulators of 452 Differentially Expressed Genes Associated With Puberty in Brahman Heifers.
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Nguyen LT, Reverter A, Cánovas A, Venus B, Anderson ST, Islas-Trejo A, Dias MM, Crawford NF, Lehnert SA, Medrano JF, Thomas MG, Moore SS, and Fortes MRS
- Abstract
The liver plays a central role in metabolism and produces important hormones. Hepatic estrogen receptors and the release of insulin-like growth factor 1 (IGF1) are critical links between liver function and the reproductive system. However, the role of liver in pubertal development is not fully understood. To explore this question, we applied transcriptomic analyses to liver samples of pre- and post-pubertal Brahman heifers and identified differentially expressed (DE) genes and genes encoding transcription factors (TFs). Differential expression of genes suggests potential biological mechanisms and pathways linking liver function to puberty. The analyses identified 452 DE genes and 82 TF with significant contribution to differential gene expression by using a regulatory impact factor metric. Brain-derived neurotrophic factor was observed as the most down-regulated gene ( P = 0.003) in post-pubertal heifers and we propose this gene influences pubertal development in Brahman heifers. Additionally, co-expression network analysis provided evidence for three TF as key regulators of liver function during pubertal development: the signal transducer and activator of transcription 6, PBX homeobox 2, and polybromo 1. Pathway enrichment analysis identified transforming growth factor-beta and Wnt signaling pathways as significant annotation terms for the list of DE genes and TF in the co-expression network. Molecular information regarding genes and pathways described in this work are important to further our understanding of puberty onset in Brahman heifers.
- Published
- 2018
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181. Individual signatures and environmental factors shape skin microbiota in healthy dogs.
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Cuscó A, Belanger JM, Gershony L, Islas-Trejo A, Levy K, Medrano JF, Sánchez A, Oberbauer AM, and Francino O
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- Animals, Bacillus classification, Bacillus genetics, Bacillus isolation & purification, Bacteria genetics, Biological Variation, Population, Environment, Female, Gammaproteobacteria classification, Gammaproteobacteria genetics, Gammaproteobacteria isolation & purification, Male, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Bacteria isolation & purification, Dogs microbiology, Microbiota genetics, Skin microbiology
- Abstract
Background: The individual, together with its environment, has been reported as the main force driving composition and structure of skin microbiota in healthy dogs. Therefore, one of the major concerns when analyzing canine skin microbiota is the likely influence of the environment. Despite the dense fur covering, certain skin diseases exhibit differential prevalence among skin sites, dog breeds, and individuals., Results: We have characterized the normal variability of dog skin microbiota in a well-controlled cohort of a large number of Golden-Labrador Retriever crossed dogs (N = 35) with similar ages, related genetic background, and a shared environment. We found that the individual drives the skin microbiota composition and structure followed by the skin site. The main bacterial classes inhabiting dog skin in this cohort are Gammaproteobacteria and Bacilli. We also detected bacteria associated to the environment on different dog skin sites that could be reflecting the different degrees of exposure of each skin site and each dog. Network analyses elucidated bacterial interactions within and between skin sites, especially in the chin, abdomen, axilla, and perianal region, with the highly shared interactions probably representing an anatomical, behavioral, or environmental component. When analyzing each skin site independently to assess host-specific factors, we found that temporality (season of birth and time spent in the kennel) affected all the skin sites and specially the inner pinna. The most abundant taxon driving this difference was Sphingomonas. We also found taxonomic differences among male and female dogs on the abdomen, axilla, and back., Conclusions: We observed a large inter-individual variability and differences among skin sites. Host-specific variables, such as temporality or sex, were also shaping skin microbiota of healthy dogs, even in an environmental homogenous cohort.
- Published
- 2017
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182. The evolution of the natural killer complex; a comparison between mammals using new high-quality genome assemblies and targeted annotation.
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Schwartz JC, Gibson MS, Heimeier D, Koren S, Phillippy AM, Bickhart DM, Smith TP, Medrano JF, and Hammond JA
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- Animals, Humans, Killer Cells, Natural metabolism, Lectins, C-Type genetics, Phylogeny, Selection, Genetic genetics, Evolution, Molecular, Genome, Mammals genetics, Molecular Sequence Annotation, Polymorphism, Genetic genetics, Receptors, Natural Killer Cell genetics, Sequence Analysis, DNA methods
- Abstract
Natural killer (NK) cells are a diverse population of lymphocytes with a range of biological roles including essential immune functions. NK cell diversity is in part created by the differential expression of cell surface receptors which modulate activation and function, including multiple subfamilies of C-type lectin receptors encoded within the NK complex (NKC). Little is known about the gene content of the NKC beyond rodent and primate lineages, other than it appears to be extremely variable between mammalian groups. We compared the NKC structure between mammalian species using new high-quality draft genome assemblies for cattle and goat; re-annotated sheep, pig, and horse genome assemblies; and the published human, rat, and mouse lemur NKC. The major NKC genes are largely in the equivalent positions in all eight species, with significant independent expansions and deletions between species, allowing us to propose a model for NKC evolution during mammalian radiation. The ruminant species, cattle and goats, have independently evolved a second KLRC locus flanked by KLRA and KLRJ, and a novel KLRH-like gene has acquired an activating tail. This novel gene has duplicated several times within cattle, while other activating receptor genes have been selectively disrupted. Targeted genome enrichment in cattle identified varying levels of allelic polymorphism between the NKC genes concentrated in the predicted extracellular ligand-binding domains. This novel recombination and allelic polymorphism is consistent with NKC evolution under balancing selection, suggesting that this diversity influences individual immune responses and may impact on differential outcomes of pathogen infection and vaccination.
- Published
- 2017
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183. Genetic Dissection of a QTL Affecting Bone Geometry.
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Sabik OL, Medrano JF, and Farber CR
- Subjects
- Alleles, Animals, Body Height genetics, Chromosome Mapping, Female, Gene Expression Regulation, Genome-Wide Association Study, Humans, Lod Score, Male, Mice, Inbred C57BL, Organ Size genetics, Polymorphism, Single Nucleotide genetics, Femur anatomy & histology, Quantitative Trait Loci genetics
- Abstract
Parameters of bone geometry such as width, length, and cross-sectional area are major determinants of bone strength. Although these traits are highly heritable, few genes influencing bone geometry have been identified. Here, we dissect a major quantitative trait locus (QTL) influencing femur size. This QTL was originally identified in an F2 cross between the C57BL/6J-hg/hg (HG) and CAST/EiJ strains and was referred to as femur length in high growth mice 2 ( Feml2 ). Feml2 was located on chromosome (Chr.) 9 at ∼20 cM. Here, we show that the HG.CAST-( D9Mit249-D9Mit133 )/Ucd congenic strain captures Feml2 In an F2 congenic cross, we fine-mapped the location of Feml2 to an ∼6 Mbp region extending from 57.3 to 63.3 Mbp on Chr. 9. We have identified candidates by mining the complete genome sequence of CAST/EiJ and through allele-specific expression (ASE) analysis of growth plates in C57BL/6J × CAST/EiJ F1 hybrids. Interestingly, we also find that the refined location of Feml2 overlaps a cluster of six independent genome-wide associations for human height. This work provides the foundation for the identification of novel genes affecting bone geometry., (Copyright © 2017 Sabik et al.)
- Published
- 2017
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184. Predicting the important enzymes in human breast milk digestion.
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Khaldi N, Vijayakumar V, Dallas DC, Guerrero A, Wickramasinghe S, Smilowitz JT, Medrano JF, Lebrilla CB, Shields DC, and German JB
- Subjects
- Gene Expression Profiling, Humans, Mass Spectrometry, Digestion, Enzymes metabolism, Milk, Human metabolism
- Abstract
Human milk is known to contain several proteases, but little is known about whether these enzymes are active, which proteins they cleave, and their relative contribution to milk protein digestion in vivo. This study analyzed the mass spectrometry-identified protein fragments found in pooled human milk by comparing their cleavage sites with the enzyme specificity patterns of an array of enzymes. The results indicate that several enzymes are actively taking part in the digestion of human milk proteins within the mammary gland, including plasmin and/or trypsin, elastase, cathepsin D, pepsin, chymotrypsin, a glutamyl endopeptidase-like enzyme, and proline endopeptidase. Two proteins were most affected by enzyme hydrolysis: β-casein and polymeric immunoglobulin receptor. In contrast, other highly abundant milk proteins such as α-lactalbumin and lactoferrin appear to have undergone no proteolytic cleavage. A peptide sequence containing a known antimicrobial peptide is released in breast milk by elastase and cathepsin D.
- Published
- 2014
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185. Multi-tissue omics analyses reveal molecular regulatory networks for puberty in composite beef cattle.
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Cánovas A, Reverter A, DeAtley KL, Ashley RL, Colgrave ML, Fortes MR, Islas-Trejo A, Lehnert S, Porto-Neto L, Rincón G, Silver GA, Snelling WM, Medrano JF, and Thomas MG
- Subjects
- Animals, Cattle physiology, Female, Fertility, Gene Regulatory Networks, Genome-Wide Association Study, Male, Pregnancy, Sexual Maturation, Transcriptome, Cattle genetics, Cattle growth & development, Gene Expression Regulation, Developmental
- Abstract
Puberty is a complex physiological event by which animals mature into an adult capable of sexual reproduction. In order to enhance our understanding of the genes and regulatory pathways and networks involved in puberty, we characterized the transcriptome of five reproductive tissues (i.e. hypothalamus, pituitary gland, ovary, uterus, and endometrium) as well as tissues known to be relevant to growth and metabolism needed to achieve puberty (i.e., longissimus dorsi muscle, adipose, and liver). These tissues were collected from pre- and post-pubertal Brangus heifers (3/8 Brahman; Bos indicus x 5/8 Angus; Bos taurus) derived from a population of cattle used to identify quantitative trait loci associated with fertility traits (i.e., age of first observed corpus luteum (ACL), first service conception (FSC), and heifer pregnancy (HPG)). In order to exploit the power of complementary omics analyses, pre- and post-puberty co-expression gene networks were constructed by combining the results from genome-wide association studies (GWAS), RNA-Seq, and bovine transcription factors. Eight tissues among pre-pubertal and post-pubertal Brangus heifers revealed 1,515 differentially expressed and 943 tissue-specific genes within the 17,832 genes confirmed by RNA-Seq analysis. The hypothalamus experienced the most notable up-regulation of genes via puberty (i.e., 204 out of 275 genes). Combining the results of GWAS and RNA-Seq, we identified 25 loci containing a single nucleotide polymorphism (SNP) associated with ACL, FSC, and (or) HPG. Seventeen of these SNP were within a gene and 13 of the genes were expressed in uterus or endometrium. Multi-tissue omics analyses revealed 2,450 co-expressed genes relative to puberty. The pre-pubertal network had 372,861 connections whereas the post-pubertal network had 328,357 connections. A sub-network from this process revealed key transcriptional regulators (i.e., PITX2, FOXA1, DACH2, PROP1, SIX6, etc.). Results from these multi-tissue omics analyses improve understanding of the number of genes and their complex interactions for puberty in cattle.
- Published
- 2014
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186. Comparison of five different RNA sources to examine the lactating bovine mammary gland transcriptome using RNA-Sequencing.
- Author
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Cánovas A, Rincón G, Bevilacqua C, Islas-Trejo A, Brenaut P, Hovey RC, Boutinaud M, Morgenthaler C, VanKlompenberg MK, Martin P, and Medrano JF
- Subjects
- Algorithms, Animals, Base Sequence, Cattle, Milk, Molecular Sequence Data, Transcription Factors metabolism, Lactation physiology, Mammary Glands, Animal physiology, RNA genetics, Sequence Analysis, RNA methods, Transcription Factors genetics, Transcriptome physiology
- Abstract
The objective of this study was to examine five different sources of RNA, namely mammary gland tissue (MGT), milk somatic cells (SC), laser microdissected mammary epithelial cells (LCMEC), milk fat globules (MFG) and antibody-captured milk mammary epithelial cells (mMEC) to analyze the bovine mammary gland transcriptome using RNA-Sequencing. Our results provide a comparison between different sampling methods (invasive and non-invasive) to define the transcriptome of mammary gland tissue and milk cells. This information will be of value to investigators in choosing the most appropriate sampling method for different research applications to study specific physiological states during lactation. One of the simplest procedures to study the transcriptome associated with milk appears to be the isolation of total RNA directly from SC or MFG released into milk during lactation. Our results indicate that the SC and MFG transcriptome are representative of MGT and LCMEC and can be used as effective and alternative samples to study mammary gland expression without the need to perform a tissue biopsy.
- Published
- 2014
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187. Genomic variation and population structure detected by single nucleotide polymorphism arrays in Corriedale, Merino and Creole sheep.
- Author
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Grasso AN, Goldberg V, Navajas EA, Iriarte W, Gimeno D, Aguilar I, Medrano JF, Rincón G, and Ciappesoni G
- Abstract
THE AIM OF THIS STUDY WAS TO INVESTIGATE THE GENETIC DIVERSITY WITHIN AND AMONG THREE BREEDS OF SHEEP: Corriedale, Merino and Creole. Sheep from the three breeds (Merino n = 110, Corriedale n = 108 and Creole n = 10) were genotyped using the Illumina Ovine SNP50 beadchip(®). Genetic diversity was evaluated by comparing the minor allele frequency (MAF) among breeds. Population structure and genetic differentiation were assessed using STRUCTURE software, principal component analysis (PCA) and fixation index (FST). Fixed markers (MAF = 0) that were different among breeds were identified as specific breed markers. Using a subset of 18,181 single nucleotide polymorphisms (SNPs), PCA and STUCTURE analysis were able to explain population stratification within breeds. Merino and Corriedale divergent lines showed high levels of polymorphism (89.4% and 86% of polymorphic SNPs, respectively) and moderate genetic differentiation (FST = 0.08) between them. In contrast, Creole had only 69% polymorphic SNPs and showed greater genetic differentiation from the other two breeds (FST = 0.17 for both breeds). Hence, a subset of molecular markers present in the OvineSNP50 is informative enough for breed assignment and population structure analysis of commercial and Creole breeds.
- Published
- 2014
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188. Molecular phylogeny and SNP variation of polar bears (Ursus maritimus), brown bears (U. arctos), and black bears (U. americanus) derived from genome sequences.
- Author
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Cronin MA, Rincon G, Meredith RW, MacNeil MD, Islas-Trejo A, Cánovas A, and Medrano JF
- Subjects
- Animals, Base Sequence, Consensus Sequence, Female, Gene Frequency, Genomic Structural Variation, Male, Pedigree, Phylogeny, Polymorphism, Single Nucleotide, Sequence Analysis, DNA, Ursidae classification, Chromosome Mapping, Genetics, Population, Genome genetics, Ursidae genetics
- Abstract
We assessed the relationships of polar bears (Ursus maritimus), brown bears (U. arctos), and black bears (U. americanus) with high throughput genomic sequencing data with an average coverage of 25× for each species. A total of 1.4 billion 100-bp paired-end reads were assembled using the polar bear and annotated giant panda (Ailuropoda melanoleuca) genome sequences as references. We identified 13.8 million single nucleotide polymorphisms (SNP) in the 3 species aligned to the polar bear genome. These data indicate that polar bears and brown bears share more SNP with each other than either does with black bears. Concatenation and coalescence-based analysis of consensus sequences of approximately 1 million base pairs of ultraconserved elements in the nuclear genome resulted in a phylogeny with black bears as the sister group to brown and polar bears, and all brown bears are in a separate clade from polar bears. Genotypes for 162 SNP loci of 336 bears from Alaska and Montana showed that the species are genetically differentiated and there is geographic population structure of brown and black bears but not polar bears.
- Published
- 2014
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189. Sequencing the transcriptome of milk production: milk trumps mammary tissue.
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Lemay DG, Hovey RC, Hartono SR, Hinde K, Smilowitz JT, Ventimiglia F, Schmidt KA, Lee JW, Islas-Trejo A, Silva PI, Korf I, Medrano JF, Barry PA, and German JB
- Subjects
- Animals, Biomarkers, Cluster Analysis, Female, Gene Expression Profiling, Gene Expression Regulation, Humans, Macaca mulatta, Mammary Glands, Animal cytology, Milk cytology, Organ Specificity genetics, Sequence Analysis, RNA, Lactation genetics, Mammary Glands, Animal metabolism, Milk metabolism, Transcriptome
- Abstract
Background: Studies of normal human mammary gland development and function have mostly relied on cell culture, limited surgical specimens, and rodent models. Although RNA extracted from human milk has been used to assay the mammary transcriptome non-invasively, this assay has not been adequately validated in primates. Thus, the objectives of the current study were to assess the suitability of lactating rhesus macaques as a model for lactating humans and to determine whether RNA extracted from milk fractions is representative of RNA extracted from mammary tissue for the purpose of studying the transcriptome of milk-producing cells., Results: We confirmed that macaque milk contains cytoplasmic crescents and that ample high-quality RNA can be obtained for sequencing. Using RNA sequencing, RNA extracted from macaque milk fat and milk cell fractions more accurately represented RNA from mammary epithelial cells (cells that produce milk) than did RNA from whole mammary tissue. Mammary epithelium-specific transcripts were more abundant in macaque milk fat, whereas adipose or stroma-specific transcripts were more abundant in mammary tissue. Functional analyses confirmed the validity of milk as a source of RNA from milk-producing mammary epithelial cells., Conclusions: RNA extracted from the milk fat during lactation accurately portrayed the RNA profile of milk-producing mammary epithelial cells in a non-human primate. However, this sample type clearly requires protocols that minimize RNA degradation. Overall, we validated the use of RNA extracted from human and macaque milk and provided evidence to support the use of lactating macaques as a model for human lactation.
- Published
- 2013
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190. RNA-seq analysis of single bovine blastocysts.
- Author
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Chitwood JL, Rincon G, Kaiser GG, Medrano JF, and Ross PJ
- Subjects
- Alleles, Animals, Cattle, Female, Male, Molecular Sequence Annotation, Polymorphism, Single Nucleotide, Reproducibility of Results, Sex Characteristics, Transcriptome, Blastocyst metabolism, Sequence Analysis, RNA
- Abstract
Background: Use of RNA-Seq presents unique benefits in terms of gene expression analysis because of its wide dynamic range and ability to identify functional sequence variants. This technology provides the opportunity to assay the developing embryo, but the paucity of biological material available from individual embryos has made this a challenging prospect., Results: We report here the first application of RNA-Seq for the analysis of individual blastocyst gene expression, SNP detection, and characterization of allele specific expression (ASE). RNA was extracted from single bovine blastocysts (n = 5), amplified, and analyzed using high-throughput sequencing. Approximately 38 million sequencing reads were generated per embryo and 9,489 known bovine genes were found to be expressed, with a high correlation of expression levels between samples (r > 0.97). Transcriptomic data was analyzed to identify SNP in expressed genes, and individual SNP were examined to characterize allele specific expression. Expressed biallelic SNP variants with allelic imbalances were observed in 473 SNP, where one allele represented between 65-95% of a variant's transcripts., Conclusions: This study represents the first application of RNA-seq technology in single bovine embryos allowing a representation of the embryonic transcriptome and the analysis of transcript sequence variation to describe specific allele expression.
- Published
- 2013
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191. Single nucleotide polymorphisms in CETP, SLC46A1, SLC19A1, CD36, BCMO1, APOA5, and ABCA1 are significant predictors of plasma HDL in healthy adults.
- Author
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Clifford AJ, Rincon G, Owens JE, Medrano JF, Moshfegh AJ, Baer DJ, and Novotny JA
- Subjects
- ATP Binding Cassette Transporter 1 genetics, Adult, Aged, Apolipoprotein A-V, Apolipoproteins A genetics, CD36 Antigens genetics, Cholesterol Ester Transfer Proteins genetics, Female, Humans, Lipoproteins, HDL genetics, Male, Middle Aged, Polymorphism, Single Nucleotide, Prognosis, Proton-Coupled Folate Transporter genetics, Reduced Folate Carrier Protein genetics, beta-Carotene 15,15'-Monooxygenase genetics, Cholesterol blood, Folic Acid Transporters genetics, Genetic Association Studies, Lipoproteins, HDL blood
- Abstract
Background: In a marker-trait association study we estimated the statistical significance of 65 single nucleotide polymorphisms (SNP) in 23 candidate genes on HDL levels of two independent Caucasian populations. Each population consisted of men and women and their HDL levels were adjusted for gender and body weight. We used a linear regression model. Selected genes corresponded to folate metabolism, vitamins B-12, A, and E, and cholesterol pathways or lipid metabolism., Methods: Extracted DNA from both the Sacramento and Beltsville populations was analyzed using an allele discrimination assay with a MALDI-TOF mass spectrometry platform. The adjusted phenotype, y, was HDL levels adjusted for gender and body weight only statistical analyses were performed using the genotype association and regression modules from the SNP Variation Suite v7., Results: Statistically significant SNP (where P values were adjusted for false discovery rate) included: CETP (rs7499892 and rs5882); SLC46A1 (rs37514694; rs739439); SLC19A1 (rs3788199); CD36 (rs3211956); BCMO1 (rs6564851), APOA5 (rs662799), and ABCA1 (rs4149267). Many prior association trends of the SNP with HDL were replicated in our cross-validation study. Significantly, the association of SNP in folate transporters (SLC46A1 rs37514694 and rs739439; SLC19A1 rs3788199) with HDL was identified in our study., Conclusions: Given recent literature on the role of niacin in the biogenesis of HDL, focus on status and metabolism of B-vitamins and metabolites of eccentric cleavage of β-carotene with lipid metabolism is exciting for future study.
- Published
- 2013
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192. Polymorphisms in genes in the SREBP1 signalling pathway and SCD are associated with milk fatty acid composition in Holstein cattle.
- Author
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Rincon G, Islas-Trejo A, Castillo AR, Bauman DE, German BJ, and Medrano JF
- Subjects
- Animals, Cattle genetics, Fatty Acids chemistry, Fatty Acids metabolism, Female, Gene Expression Regulation physiology, Polymorphism, Genetic, Polymorphism, Single Nucleotide, Signal Transduction physiology, Cattle metabolism, Milk chemistry, Stearoyl-CoA Desaturase genetics, Stearoyl-CoA Desaturase metabolism, Sterol Regulatory Element Binding Protein 1 genetics, Sterol Regulatory Element Binding Protein 1 metabolism
- Abstract
Genes in the sterol regulatory element-binding protein-1 (SREBP1) pathway play a central role in regulation of milk fat synthesis, especially the de-novo synthesis of saturated fatty acids. SCD, a SREBP-responsive gene, is the key enzyme in the synthesis of monounsaturated fatty acids in the mammary gland. In the present study, we discovered SNP in candidate genes associated with this signalling pathway and SCD to identify genetic markers that can be used for genetic and metabolically directed selection in cattle. We resequenced six candidate genes in the SREBP1 pathway (SREBP1, SCAP, INSIG1, INSIG2, MBTPS1, MBTPS2) and two genes for SCD (SCD1 and SCD5) and discovered 47 Tag SNP that were used in a marker-trait association study. Milk and blood samples were collected from Holstein cows in their 1st or 2nd parity at 100-150 days of lactation. Individual fatty acids from C4 to C20, saturated fatty acid (SFA) content, monounsaturated fatty acid content, polyunsaturated fatty acid content and desaturase indexes were measured and used to perform the asociation analysis. Polymorphisms in the SCD5 and INSIG2 genes were the most representative markers associated with SFA/unsaturated fatty acid (UFA) ratio in milk. The analysis of desaturation activity determined that markers in the SCD1 and SCD5 genes showed the most significant effects. DGAT1 K232A marker was included in the study to examine the effect of this marker on the variation of milk fatty acids in our Holstein population. The percentage of variance explained by DGAT1 in the analysis was only 6% of SFA/UFA ratio. Milk fat depression was observed in one of the dairy herds and in this particular dairy one SNP in the SREBP1 gene (rs41912290) accounted for 40% of the phenotypic variance. Our results provide detailed SNP information for key genes in the SREBP1 signalling pathway and SCD that can be used to change milk fat composition by marker-assisted breeding to meet consumer demands regarding human health, as well as furthering understanding of technological aspects of cows' milk.
- Published
- 2012
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193. Transcriptional profiling of bovine milk using RNA sequencing.
- Author
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Wickramasinghe S, Rincon G, Islas-Trejo A, and Medrano JF
- Subjects
- Animals, Cattle, Epithelial Cells enzymology, Peptide Hydrolases genetics, Peptide Hydrolases metabolism, Proteasome Endopeptidase Complex genetics, Proteasome Endopeptidase Complex metabolism, RNA isolation & purification, Sequence Analysis, RNA, Ubiquitin genetics, Ubiquitin metabolism, Epithelial Cells metabolism, Milk cytology, Transcriptome
- Abstract
Background: Cow milk is a complex bioactive fluid consumed by humans beyond infancy. Even though the chemical and physical properties of cow milk are well characterized, very limited research has been done on characterizing the milk transcriptome. This study performs a comprehensive expression profiling of genes expressed in milk somatic cells of transition (day 15), peak (day 90) and late (day 250) lactation Holstein cows by RNA sequencing. Milk samples were collected from Holstein cows at 15, 90 and 250 days of lactation, and RNA was extracted from the pelleted milk cells. Gene expression analysis was conducted by Illumina RNA sequencing. Sequence reads were assembled and analyzed in CLC Genomics Workbench. Gene Ontology (GO) and pathway analysis were performed using the Blast2GO program and GeneGo application of MetaCore program., Results: A total of 16,892 genes were expressed in transition lactation, 19,094 genes were expressed in peak lactation and 18,070 genes were expressed in late lactation. Regardless of the lactation stage approximately 9,000 genes showed ubiquitous expression. Genes encoding caseins, whey proteins and enzymes in lactose synthesis pathway showed higher expression in early lactation. The majority of genes in the fat metabolism pathway had high expression in transition and peak lactation milk. Most of the genes encoding for endogenous proteases and enzymes in ubiquitin-proteasome pathway showed higher expression along the course of lactation., Conclusions: This is the first study to describe the comprehensive bovine milk transcriptome in Holstein cows. The results revealed that 69% of NCBI Btau 4.0 annotated genes are expressed in bovine milk somatic cells. Most of the genes were ubiquitously expressed in all three stages of lactation. However, a fraction of the milk transcriptome has genes devoted to specific functions unique to the lactation stage. This indicates the ability of milk somatic cells to adapt to different molecular functions according to the biological need of the animal. This study provides a valuable insight into the biology of lactation in the cow, as well as many avenues for future research on the bovine lactome.
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- 2012
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194. Comparison of buccal and blood-derived canine DNA, either native or whole genome amplified, for array-based genome-wide association studies.
- Author
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Rincon G, Tengvall K, Belanger JM, Lagoutte L, Medrano JF, André C, Thomas A, Lawley CT, Hansen MS, Lindblad-Toh K, and Oberbauer AM
- Abstract
Background: The availability of array-based genotyping platforms for single nucleotide polymorphisms (SNPs) for the canine genome has expanded the opportunities to undertake genome-wide association (GWA) studies to identify the genetic basis for Mendelian and complex traits. Whole blood as the source of high quality DNA is undisputed but often proves impractical for collection of the large numbers of samples necessary to discover the loci underlying complex traits. Further, many countries prohibit the collection of blood from dogs unless medically necessary thereby restricting access to critical control samples from healthy dogs. Alternate sources of DNA, typically from buccal cytobrush extractions, while convenient, have been suggested to have low yield and perform poorly in GWA. Yet buccal cytobrushes provide a cost-effective means of collecting DNA, are readily accepted by dog owners, and represent a large resource base in many canine genetics laboratories. To increase the DNA quantities, whole genome amplification (WGA) can be performed. Thus, the present study assessed the utility of buccal-derived DNA as well as whole genome amplification in comparison to blood samples for use on the most recent iteration of the canine HD SNP array (Illumina)., Findings: In both buccal and blood samples, whether whole genome amplified or not, 97% of the samples had SNP call rates in excess of 80% indicating that the vast majority of the SNPs would be suitable to perform association studies regardless of the DNA source. Similarly, there were no significant differences in marker intensity measurements between buccal and blood samples for copy number variations (CNV) analysis., Conclusions: All DNA samples assayed, buccal or blood, native or whole genome amplified, are appropriate for use in array-based genome-wide association studies. The concordance between subsets of dogs for which both buccal and blood samples, or those samples whole genome amplified, was shown to average >99%. Thus, the two DNA sources were comparable in the generation of SNP genotypes and intensity values to estimate structural variation indicating the utility for the use of buccal cytobrush samples and the reliability of whole genome amplification for genome-wide association and CNV studies.
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- 2011
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195. Transcriptome profiling of bovine milk oligosaccharide metabolism genes using RNA-sequencing.
- Author
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Wickramasinghe S, Hua S, Rincon G, Islas-Trejo A, German JB, Lebrilla CB, and Medrano JF
- Subjects
- Animals, Female, Fucose metabolism, Gene Expression Regulation, Glycosylation, Lactation genetics, Mammals genetics, Metabolic Networks and Pathways genetics, Molecular Sequence Annotation, N-Acetylneuraminic Acid metabolism, Cattle genetics, Gene Expression Profiling, Milk chemistry, Oligosaccharides genetics, Oligosaccharides metabolism, Sequence Analysis, RNA methods
- Abstract
This study examines the genes coding for enzymes involved in bovine milk oligosaccharide metabolism by comparing the oligosaccharide profiles with the expressions of glycosylation-related genes. Fresh milk samples (n = 32) were collected from four Holstein and Jersey cows at days 1, 15, 90 and 250 of lactation and free milk oligosaccharide profiles were analyzed. RNA was extracted from milk somatic cells at days 15 and 250 of lactation (n = 12) and gene expression analysis was conducted by RNA-Sequencing. A list was created of 121 glycosylation-related genes involved in oligosaccharide metabolism pathways in bovine by analyzing the oligosaccharide profiles and performing an extensive literature search. No significant differences were observed in either oligosaccharide profiles or expressions of glycosylation-related genes between Holstein and Jersey cows. The highest concentrations of free oligosaccharides were observed in the colostrum samples and a sharp decrease was observed in the concentration of free oligosaccharides on day 15, followed by progressive decrease on days 90 and 250. Ninety-two glycosylation-related genes were expressed in milk somatic cells. Most of these genes exhibited higher expression in day 250 samples indicating increases in net glycosylation-related metabolism in spite of decreases in free milk oligosaccharides in late lactation milk. Even though fucosylated free oligosaccharides were not identified, gene expression indicated the likely presence of fucosylated oligosaccharides in bovine milk. Fucosidase genes were expressed in milk and a possible explanation for not detecting fucosylated free oligosaccharides is the degradation of large fucosylated free oligosaccharides by the fucosidases. Detailed characterization of enzymes encoded by the 92 glycosylation-related genes identified in this study will provide the basic knowledge for metabolic network analysis of oligosaccharides in mammalian milk. These candidate genes will guide the design of a targeted breeding strategy to optimize the content of beneficial oligosaccharides in bovine milk.
- Published
- 2011
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196. Characterization of bacteria in biopsies of colon and stools by high throughput sequencing of the V2 region of bacterial 16S rRNA gene in human.
- Author
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Momozawa Y, Deffontaine V, Louis E, and Medrano JF
- Subjects
- Adult, Bacteria classification, Biopsy, DNA, Bacterial genetics, DNA, Bacterial isolation & purification, Female, Humans, Male, Middle Aged, Organ Specificity, Polymerase Chain Reaction, Reproducibility of Results, Sequence Analysis, RNA, Bacteria genetics, Colon microbiology, Colon pathology, Feces microbiology, High-Throughput Nucleotide Sequencing methods, RNA, Bacterial genetics, RNA, Ribosomal, 16S genetics
- Abstract
Background: The characterization of the human intestinal microflora and their interactions with the host have been identified as key components in the study of intestinal disorders such as inflammatory bowel diseases. High-throughput sequencing has enabled culture-independent studies to deeply analyze bacteria in the gut. It is possible with this technology to systematically analyze links between microbes and the genetic constitution of the host, such as DNA polymorphisms and methylation, and gene expression., Methods and Findings: In this study the V2 region of the bacterial 16S ribosomal RNA (rRNA) gene using 454 pyrosequencing from seven anatomic regions of human colon and two types of stool specimens were analyzed. The study examined the number of reads needed to ascertain differences between samples, the effect of DNA extraction procedures and PCR reproducibility, and differences between biopsies and stools in order to design a large scale systematic analysis of gut microbes. It was shown (1) that sequence coverage lower than 1,000 reads influenced quantitative and qualitative differences between samples measured by UniFrac distances. Distances between samples became stable after 1,000 reads. (2) Difference of extracted bacteria was observed between the two DNA extraction methods. In particular, Firmicutes Bacilli were not extracted well by one method. (3) Quantitative and qualitative difference in bacteria from ileum to rectum colon were not observed, but there was a significant positive trend between distances within colon and quantitative differences. Between sample type, biopsies or stools, quantitative and qualitative differences were observed., Conclusions: Results of human colonic bacteria analyzed using high-throughput sequencing were highly dependent on the experimental design, especially the number of sequence reads, DNA extraction method, and sample type.
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- 2011
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197. SNP discovery in the bovine milk transcriptome using RNA-Seq technology.
- Author
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Cánovas A, Rincon G, Islas-Trejo A, Wickramasinghe S, and Medrano JF
- Subjects
- Animals, DNA, Complementary genetics, DNA, Complementary metabolism, Female, Genotype, Cattle genetics, Gene Expression Profiling methods, Milk metabolism, Polymorphism, Single Nucleotide, Sequence Analysis, DNA methods
- Abstract
High-throughput sequencing of RNA (RNA-Seq) was developed primarily to analyze global gene expression in different tissues. However, it also is an efficient way to discover coding SNPs. The objective of this study was to perform a SNP discovery analysis in the milk transcriptome using RNA-Seq. Seven milk samples from Holstein cows were analyzed by sequencing cDNAs using the Illumina Genome Analyzer system. We detected 19,175 genes expressed in milk samples corresponding to approximately 70% of the total number of genes analyzed. The SNP detection analysis revealed 100,734 SNPs in Holstein samples, and a large number of those corresponded to differences between the Holstein breed and the Hereford bovine genome assembly Btau4.0. The number of polymorphic SNPs within Holstein cows was 33,045. The accuracy of RNA-Seq SNP discovery was tested by comparing SNPs detected in a set of 42 candidate genes expressed in milk that had been resequenced earlier using Sanger sequencing technology. Seventy of 86 SNPs were detected using both RNA-Seq and Sanger sequencing technologies. The KASPar Genotyping System was used to validate unique SNPs found by RNA-Seq but not observed by Sanger technology. Our results confirm that analyzing the transcriptome using RNA-Seq technology is an efficient and cost-effective method to identify SNPs in transcribed regions. This study creates guidelines to maximize the accuracy of SNP discovery and prevention of false-positive SNP detection, and provides more than 33,000 SNPs located in coding regions of genes expressed during lactation that can be used to develop genotyping platforms to perform marker-trait association studies in Holstein cattle.
- Published
- 2010
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198. Conserved role of unc-79 in ethanol responses in lightweight mutant mice.
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Speca DJ, Chihara D, Ashique AM, Bowers MS, Pierce-Shimomura JT, Lee J, Rabbee N, Speed TP, Gularte RJ, Chitwood J, Medrano JF, Liao M, Sonner JM, Eger EI 2nd, Peterson AS, and McIntire SL
- Subjects
- Animals, Caenorhabditis elegans genetics, Caenorhabditis elegans growth & development, Caenorhabditis elegans metabolism, Caenorhabditis elegans physiology, Caenorhabditis elegans Proteins genetics, Caenorhabditis elegans Proteins metabolism, Female, Ion Channels genetics, Ion Channels metabolism, Male, Membrane Proteins, Mice genetics, Mice growth & development, Mice physiology, Mice, Inbred C57BL, Motor Activity, Body Weight, Ethanol metabolism, Mice metabolism, Mutation, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism
- Abstract
The mechanisms by which ethanol and inhaled anesthetics influence the nervous system are poorly understood. Here we describe the positional cloning and characterization of a new mouse mutation isolated in an N-ethyl-N-nitrosourea (ENU) forward mutagenesis screen for animals with enhanced locomotor activity. This allele, Lightweight (Lwt), disrupts the homolog of the Caenorhabditis elegans (C. elegans) unc-79 gene. While Lwt/Lwt homozygotes are perinatal lethal, Lightweight heterozygotes are dramatically hypersensitive to acute ethanol exposure. Experiments in C. elegans demonstrate a conserved hypersensitivity to ethanol in unc-79 mutants and extend this observation to the related unc-80 mutant and nca-1;nca-2 double mutants. Lightweight heterozygotes also exhibit an altered response to the anesthetic isoflurane, reminiscent of unc-79 invertebrate mutant phenotypes. Consistent with our initial mapping results, Lightweight heterozygotes are mildly hyperactive when exposed to a novel environment and are smaller than wild-type animals. In addition, Lightweight heterozygotes exhibit increased food consumption yet have a leaner body composition. Interestingly, Lightweight heterozygotes voluntarily consume more ethanol than wild-type littermates. The acute hypersensitivity to and increased voluntary consumption of ethanol observed in Lightweight heterozygous mice in combination with the observed hypersensitivity to ethanol in C. elegans unc-79, unc-80, and nca-1;nca-2 double mutants suggests a novel conserved pathway that might influence alcohol-related behaviors in humans., Competing Interests: EIE is a paid consultant of Baxter Healthcare.
- Published
- 2010
- Full Text
- View/download PDF
199. Serious limitations of the QTL/microarray approach for QTL gene discovery.
- Author
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Verdugo RA, Farber CR, Warden CH, and Medrano JF
- Subjects
- Animals, Genes, Humans, Mice, Mice, Congenic genetics, Oligonucleotide Array Sequence Analysis economics, Oligonucleotide Array Sequence Analysis methods, Quantitative Trait Loci
- Abstract
Background: It has been proposed that the use of gene expression microarrays in nonrecombinant parental or congenic strains can accelerate the process of isolating individual genes underlying quantitative trait loci (QTL). However, the effectiveness of this approach has not been assessed., Results: Thirty-seven studies that have implemented the QTL/microarray approach in rodents were reviewed. About 30% of studies showed enrichment for QTL candidates, mostly in comparisons between congenic and background strains. Three studies led to the identification of an underlying QTL gene. To complement the literature results, a microarray experiment was performed using three mouse congenic strains isolating the effects of at least 25 biometric QTL. Results show that genes in the congenic donor regions were preferentially selected. However, within donor regions, the distribution of differentially expressed genes was homogeneous once gene density was accounted for. Genes within identical-by-descent (IBD) regions were less likely to be differentially expressed in chromosome 2, but not in chromosomes 11 and 17. Furthermore, expression of QTL regulated in cis (cis eQTL) showed higher expression in the background genotype, which was partially explained by the presence of single nucleotide polymorphisms (SNP)., Conclusions: The literature shows limited successes from the QTL/microarray approach to identify QTL genes. Our own results from microarray profiling of three congenic strains revealed a strong tendency to select cis-eQTL over trans-eQTL. IBD regions had little effect on rate of differential expression, and we provide several reasons why IBD should not be used to discard eQTL candidates. In addition, mismatch probes produced false cis-eQTL that could not be completely removed with the current strains genotypes and low probe density microarrays. The reviewed studies did not account for lack of coverage from the platforms used and therefore removed genes that were not tested. Together, our results explain the tendency to report QTL candidates as differentially expressed and indicate that the utility of the QTL/microarray as currently implemented is limited. Alternatives are proposed that make use of microarray data from multiple experiments to overcome the outlined limitations.
- Published
- 2010
- Full Text
- View/download PDF
200. Segregation analysis of a sex ratio distortion locus in congenic mice.
- Author
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Casellas J, Farber CR, Verdugo RA, and Medrano JF
- Subjects
- Animals, Crosses, Genetic, Female, Genes, Dominant, Male, Mice, Mice, Congenic, Mice, Inbred C57BL, Pedigree, Genetic Loci, Models, Genetic, Sex Ratio
- Abstract
The congenic HG.CAST-(D17Mit196-D17Mit190) (HQ17(hg/hg)) mouse strain showed a significant departure on the expected 50%/50% offspring sex ratio in more than 2400 progeny (55.7% females). The entire pedigree file included data from 13 nonoverlapping purebred generations and an F(2) cross with the C57BL/6J inbred strain. Offspring sex ratio data were analyzed on the basis of 40 purebred HQ17(hg)(/hg) sires and 29 F(1) HQ17(hg)(/hg) x B6 sires under a Bayesian Binomial segregation model accounting for 4 different autosomal inheritance models of gene action (i.e., additive, dominance, recessive, and overdominance) and X-linked and Y-linked loci. For each model, the segregation effect was evaluated as a single regression coefficient for all sires or assuming 2 independent regression coefficients accounting for offspring sex ratio departures in purebred and F(1) sires, respectively. The deviance information criterion clearly favored the autosomal dominance model with different regression coefficients for the 2 groups of sires. Under this model, the dominance effect increased the percentage of female offspring by 4.3% (HQ17(hg)(/hg) purebred sires) and 8.2% (F(1) sires) with the highest posterior density regions ranging from 0.5% to 10.6% and from 1.3% to 14.4%, respectively. This article provides significant evidence of genetic determinism for sex ratio distortion in the HQ17(hg)(/hg) strain and develops new analytical tools to perform segregation studies on dichotomous traits.
- Published
- 2010
- Full Text
- View/download PDF
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