20 results on '"DeLay, Bridget D."'
Search Results
2. Xenopus: leaping forward in kidney organogenesis
- Author
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Krneta-Stankic, Vanja, DeLay, Bridget D., and Miller, Rachel K.
- Subjects
Health - Abstract
While kidney donations stagnate, the number of people in need of kidney transplants continues to grow. Although transplanting culture-grown organs is years away, pursuing the engineering of the kidney de novo is a valid means of closing the gap between the supply and demand of kidneys for transplantation. The structural organization of a mouse kidney is similar to that of humans. Therefore, mice have traditionally served as the primary model system for the study of kidney development. The mouse is an ideal model organism for understanding the complexity of the human kidney. Nonetheless, the elaborate structure of the mammalian kidney makes the discovery of new therapies based on de novo engineered kidneys more challenging. In contrast to mammals, amphibians have a kidney that is anatomically less complex and develops faster. Given that analogous genetic networks regulate the development of mammalian and amphibian nephric organs, using embryonic kidneys of Xenopus laevis (African clawed frog) to analyze inductive cell signaling events and morphogenesis has many advantages. Pioneering work that led to the ability to generate kidney organoids from embryonic cells was carried out in Xenopus. In this review, we discuss how Xenopus can be utilized to compliment the work performed in mammalian systems to understand kidney development., Author(s): Vanja Krneta-Stankic [sup.1] [sup.2] , Bridget D. DeLay [sup.1] , Rachel K. Miller [sup.1] [sup.2] [sup.3] [sup.4] Author Affiliations: (1) 0000 0000 9206 2401grid.267308.8Department of Pediatrics, Pediatric Research Center, [...]
- Published
- 2017
- Full Text
- View/download PDF
3. A comparative study of cellular diversity between the Xenopus pronephric and mouse metanephric nephron
- Author
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Corkins, Mark E., primary, Achieng, MaryAnne, additional, DeLay, Bridget D., additional, Krneta-Stankic, Vanja, additional, Cain, Margo P., additional, Walker, Brandy L., additional, Chen, Jichao, additional, Lindström, Nils O., additional, and Miller, Rachel K., additional
- Published
- 2022
- Full Text
- View/download PDF
4. Tissue-Targeted CRISPR–Cas9-Mediated Genome Editing of Multiple Homeologs in F0-Generation Xenopus laevis Embryos
- Author
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Corkins, Mark E., primary, DeLay, Bridget D., additional, and Miller, Rachel K., additional
- Published
- 2021
- Full Text
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5. A comparative study of cellular diversity between the Xenopuspronephric and mouse metanephric nephron
- Author
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Corkins, Mark E., Achieng, MaryAnne, DeLay, Bridget D., Krneta-Stankic, Vanja, Cain, Margo P., Walker, Brandy L., Chen, Jichao, Lindström, Nils O., and Miller, Rachel K.
- Abstract
The kidney is an essential organ that ensures bodily fluid homeostasis and removes soluble waste products from the organism. Nephrons, the functional units of the kidney, comprise a blood filter, the glomerulus or glomus, and an epithelial tubule that processes the filtrate from the blood or coelom and selectively reabsorbs solutes, such as sugars, proteins, ions, and water, leaving waste products to be eliminated in the urine. Genes coding for transporters are segmentally expressed, enabling the nephron to sequentially process the filtrate. The Xenopusembryonic kidney, the pronephros, which consists of a single large nephron, has served as a valuable model to identify genes involved in nephron formation and patterning. Therefore, the developmental patterning program that generates these segments is of great interest. Prior work has defined the gene expression profiles of Xenopusnephron segments via in situhybridization strategies, but a comprehensive understanding of the cellular makeup of the pronephric kidney remains incomplete. Here, we carried out single-cell mRNA sequencing of the functional Xenopuspronephric nephron and evaluated its cellular composition through comparative analyses with previous Xenopusstudies and single-cell mRNA sequencing of the adult mouse kidney. This study reconstructs the cellular makeup of the pronephric kidney and identifies conserved cells, segments, and associated gene expression profiles. Thus, our data highlight significant conservation in podocytes, proximal and distal tubule cells, and divergence in cellular composition underlying the capacity of each nephron to remove wastes in the form of urine, while emphasizing the Xenopuspronephros as a model for physiology and disease.
- Published
- 2023
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- View/download PDF
6. Parameters Affecting Continuous In Vitro Culture of Treponema pallidum Strains
- Author
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Edmondson, Diane G., primary, DeLay, Bridget D., additional, Kowis, Lindsay E., additional, and Norris, Steven J., additional
- Published
- 2021
- Full Text
- View/download PDF
7. Dynamin Binding Protein Is Required for Xenopus laevis Kidney Development
- Author
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DeLay, Bridget D., primary, Baldwin, Tanya A., additional, and Miller, Rachel K., additional
- Published
- 2019
- Full Text
- View/download PDF
8. Transgenic Xenopus laevis line for in vivo labeling of nephrons within the kidney
- Author
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Corkins, Mark E., Hanania, Hannah L., Krneta-Stankic, Vanja, DeLay, Bridget D., Pearl, Esther J., Lee, Moonsup, Ji, Hong, Davidson, Alan J., Horb, Marko E., Miller, Rachel K., Corkins, Mark E., Hanania, Hannah L., Krneta-Stankic, Vanja, DeLay, Bridget D., Pearl, Esther J., Lee, Moonsup, Ji, Hong, Davidson, Alan J., Horb, Marko E., and Miller, Rachel K.
- Abstract
© The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Genes 9 (2018): 197, doi:10.3390/genes9040197., Xenopus laevis embryos are an established model for studying kidney development. The nephron structure and genetic pathways that regulate nephrogenesis are conserved between Xenopus and humans, allowing for the study of human disease-causing genes. Xenopus embryos are also amenable to large-scale screening, but studies of kidney disease-related genes have been impeded because assessment of kidney development has largely been limited to examining fixed embryos. To overcome this problem, we have generated a transgenic line that labels the kidney. We characterize this cdh17:eGFP line, showing green fluorescent protein (GFP) expression in the pronephric and mesonephric kidneys and colocalization with known kidney markers. We also demonstrate the feasibility of live imaging of embryonic kidney development and the use of cdh17:eGFP as a kidney marker for secretion assays. Additionally, we develop a new methodology to isolate and identify kidney cells for primary culture. We also use morpholino knockdown of essential kidney development genes to establish that GFP expression enables observation of phenotypes, previously only described in fixed embryos. Taken together, this transgenic line will enable primary kidney cell culture and live imaging of pronephric and mesonephric kidney development. It will also provide a simple means for high-throughput screening of putative human kidney disease-causing genes., These studies were supported by a postdoctoral fellowship from the National Kidney Foundation (FLB1628 to R.K.M), a National Institutes of Health (NIH) KO1 grant (K01DK092320 to R.K.M.), and startup funding from UTHealth McGovern Medical School’s Department of Pediatrics (to R.K.M.).
- Published
- 2018
9. Targeted knockout of lhx1 via CRISPR/Cas9 gene editing in the Xenopus laevis kidney
- Author
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DeLay, Bridget D., Corkins, Mark E., Hanania, Hannah L., Salanga, Matthew C., Deng, Jian Min, Sudou, Norihiro, Taira, Masanori, Horb, Marko E., Miller, Rachel K., DeLay, Bridget D., Corkins, Mark E., Hanania, Hannah L., Salanga, Matthew C., Deng, Jian Min, Sudou, Norihiro, Taira, Masanori, Horb, Marko E., and Miller, Rachel K.
- Abstract
Author Posting. © The Author(s), 2017. This is the author's version of the work. It is posted here by permission of Genetics Society of America for personal use, not for redistribution. The definitive version was published in Genetics 208 (2018): 673-686, doi:10.1534/genetics.117.300468., Studying genes involved in organogenesis is often difficult because many of these genes are also essential for early development. The allotetraploid frog, Xenopus laevis, is commonly used to study developmental processes, but because of the presence of two homeologs for many genes, it has been difficult to use as a genetic model. Few studies have successfully used CRISPR in amphibians, and currently there is no tissue-targeted knockout strategy described in Xenopus. The goal of this study is to determine whether CRISPR/Cas9-mediated gene knockout can be targeted to the Xenopus kidney without perturbing essential early gene function. We demonstrate that targeting CRISPR gene editing to the kidney and the eye of F0 embryos is feasible. Our study shows that knockout of both homeologs of lhx1 results in the disruption of kidney development and function but does not lead to early developmental defects. Therefore, targeting of CRISPR to the kidney may not be necessary to bypass the early developmental defects reported upon disruption of Lhx1 protein expression or function by morpholinos, antisense RNA, or dominant negative constructs. We also establish a control for CRISPR in Xenopus by editing a gene (slc45a2) that when knocked out results in albinism without altering kidney development. This study establishes the feasibility of tissue-specific gene knockout in Xenopus, providing a cost effective and efficient method for assessing the roles of genes implicated in developmental abnormalities that is amenable to high-throughput gene or drug screening techniques., These studies were supported by a National Institutes of Health (NIH) KO1 grant (K01DK092320 to R.K.M.), startup funding from the Department of Pediatrics 424 Pediatric Research Center at the University of Texas McGovern Medical School (to R.K.M.), an NIH National Xenopus Resource Center grant (P40OD010997 to M.E.H.), and an NIH R01 grant (R01HD084409 to M.E.H.).
- Published
- 2018
10. Dynamin binding protein is required for Xenopus laevis kidney development
- Author
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DeLay, Bridget D., primary, Baldwin, Tanya A., additional, and Miller, Rachel K., additional
- Published
- 2018
- Full Text
- View/download PDF
11. Tissue-Specific Gene Inactivation in Xenopus laevis: Knockout of lhx1 in the Kidney with CRISPR/Cas9
- Author
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DeLay, Bridget D, primary, Corkins, Mark E, additional, Hanania, Hannah L, additional, Salanga, Matthew, additional, Deng, Jian Min, additional, Sudou, Norihiro, additional, Taira, Masanori, additional, Horb, Marko E, additional, and Miller, Rachel K, additional
- Published
- 2018
- Full Text
- View/download PDF
12. Technique to Target Microinjection to the Developing Xenopus Kidney
- Author
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DeLay, Bridget D., primary, Krneta-Stankic, Vanja, primary, and Miller, Rachel K., primary
- Published
- 2016
- Full Text
- View/download PDF
13. Xenopus: leaping forward in kidney organogenesis
- Author
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Krneta-Stankic, Vanja, primary, DeLay, Bridget D., additional, and Miller, Rachel K., additional
- Published
- 2016
- Full Text
- View/download PDF
14. The Importance of Gut Symbionts in the Development of the Brown Marmorated Stink Bug, Halyomorpha halys (Stål)
- Author
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Taylor, Christopher M., primary, Coffey, Peter L., additional, DeLay, Bridget D., additional, and Dively, Galen P., additional
- Published
- 2014
- Full Text
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15. Tissue-Specific Gene Inactivation in Xenopus laevis: Knockout of lhx1in the Kidney with CRISPR/Cas9
- Author
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DeLay, Bridget D, Corkins, Mark E, Hanania, Hannah L, Salanga, Matthew, Deng, Jian Min, Sudou, Norihiro, Taira, Masanori, Horb, Marko E, and Miller, Rachel K
- Abstract
Xenopus laevisis a classic developmental model, but its allotetraploid genome has limited our ability to perform genetic manipulations. The advance of...Studying genes involved in organogenesis is often difficult because many of these genes are also essential for early development. The allotetraploid frog, Xenopus laevis, is commonly used to study developmental processes, but because of the presence of two homeologs for many genes, it has been difficult to use as a genetic model. Few studies have successfully used CRISPR in amphibians, and currently there is no tissue-targeted knockout strategy described in Xenopus. The goal of this study is to determine whether CRISPR/Cas9-mediated gene knockout can be targeted to the Xenopuskidney without perturbing essential early gene function. We demonstrate that targeting CRISPR gene editing to the kidney and the eye of F0 embryos is feasible. Our study shows that knockout of both homeologs of lhx1results in the disruption of kidney development and function but does not lead to early developmental defects. Therefore, targeting of CRISPR to the kidney may not be necessary to bypass the early developmental defects reported upon disruption of Lhx1 protein expression or function by morpholinos, antisense RNA, or dominant negative constructs. We also establish a control for CRISPR in Xenopusby editing a gene (slc45a2) that when knocked out results in albinism without altering kidney development. This study establishes the feasibility of tissue-specific gene knockout in Xenopus, providing a cost-effective and efficient method for assessing the roles of genes implicated in developmental abnormalities that is amenable to high-throughput gene or drug screening techniques.
- Published
- 2018
- Full Text
- View/download PDF
16. The Importance of Gut Symbionts in the Development of the Brown Marmorated Stink Bug, Halyomorpha halys (Stål).
- Author
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Taylor, Christopher M., Coffey, Peter L., DeLay, Bridget D., and Dively, Galen P.
- Subjects
STINKBUGS ,INTRODUCED insects ,CONTROL of agricultural pests & diseases ,INSECT-plant symbiosis ,CROP management ,SUSTAINABLE agriculture ,DEVELOPMENTAL biology - Abstract
The invasive brown marmorated stink bug, Halyomorpha halys (Stål), has become a severe agricultural pest and nuisance problem since its introduction in the U.S. Research is being conducted to understand its biology and to find management solutions. Its symbiotic relationship with gut symbionts is one aspect of its biology that is not understood. In the family Pentatomidae, the reliance on gut symbionts for successful development seems to vary depending on the species of stink bug. This research assessed the role of gut symbionts in the development, survivorship, and fecundity of H. halys. We compared various fitness parameters of nymphs and adults reared from surface sterilized and untreated egg masses during two consecutive generations under laboratory conditions. Results provided direct evidence that H. halys is negatively impacted by the prevention of vertical transmission of its gut symbionts and that this impact is significant in the first generation and manifests dramatically in the subsequent generation. Developmental time and survivorship of treated cohorts in the first generation were significantly affected during third instar development through to the adult stage. Adults from the sterilized treatment group exhibited longer pre-oviposition periods, produced fewer egg masses, had significantly smaller clutch sizes, and the hatch rate and survivorship of those eggs were significantly reduced. Observations following hatch of surface sterilized eggs also revealed significant effects on wandering behavior of the first instars. The second generation progeny from adults of the sterilized cohorts showed significantly lower survival to adulthood, averaging only 0.3% compared to 20.8% for the control cohorts. Taken together, results demonstrate that H. halys is heavily impacted by deprival of its gut symbionts. Given the economic status of this invasive pest, further investigations may lead to management tactics that disrupt this close symbiotic relationship in the biology of H. halys. [ABSTRACT FROM AUTHOR]
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- 2014
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17. Tissue-Targeted CRISPR–Cas9-Mediated Genome Editing of Multiple Homeologs in F0-Generation Xenopus laevisEmbryos
- Author
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Corkins, Mark E., DeLay, Bridget D., and Miller, Rachel K.
- Abstract
Xenopus laevisfrogs are a powerful developmental model that enables studies combining classical embryology and molecular manipulation. Because of the large embryo size, ease of microinjection, and ability to target tissues through established fate maps, X. laevishas become the predominant amphibian research model. Given that their allotetraploid genome has complicated the generation of gene knockouts, strategies need to be established for efficient mutagenesis of multiple homeologs to evaluate gene function. Here we describe a protocol to use CRISPR–Cas9-mediated genome editing to target either single alleles or multiple alloalleles in F0X. laevisembryos. A single-guide RNA (sgRNA) is designed to target a specific DNA sequence encoding a critical protein domain. To mutagenize a gene with two alloalleles, the sgRNA is designed against a sequence that is common to both homeologs. This sgRNA, along with the Cas9 protein, is microinjected into the zygote to disrupt the genomic sequences in the whole embryo or into a specific blastomere for tissue-targeted effects. Error-prone repair of CRISPR–Cas9-generated DNA double-strand breaks leads to insertions and deletions creating mosaic gene lesions within the embryos. The genomic DNA isolated from each mosaic F0embryo is sequenced, and software is applied to assess the nature of the mutations generated and degree of mosaicism. This protocol enables the knockout of genes within the whole embryo or in specific tissues in F0X. laevisembryos to facilitate the evaluation of resulting phenotypes.
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- 2022
- Full Text
- View/download PDF
18. Parameters Affecting Continuous In VitroCulture of Treponema pallidumStrains
- Author
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Edmondson, Diane G., DeLay, Bridget D., Kowis, Lindsay E., and Norris, Steven J.
- Abstract
Syphilis is caused by the bacterium Treponema pallidumsubsp. pallidum. Until recently, this pathogen could only be maintained through infection of rabbits or other animals, making study of this important human pathogen challenging and costly. T. pallidumsubsp. pallidumhas now been successfully cultured for over 3 years in a tissue culture system using a medium called TpCM-2.
- Published
- 2021
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19. Tissue-Targeted CRISPR-Cas9-Mediated Genome Editing of Multiple Homeologs in F 0 -Generation Xenopus laevis Embryos.
- Author
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Corkins ME, DeLay BD, and Miller RK
- Subjects
- Animals, CRISPR-Associated Protein 9 genetics, RNA, Guide, CRISPR-Cas Systems genetics, Xenopus laevis genetics, CRISPR-Cas Systems, Gene Editing methods
- Abstract
Xenopus laevis frogs are a powerful developmental model that enables studies combining classical embryology and molecular manipulation. Because of the large embryo size, ease of microinjection, and ability to target tissues through established fate maps, X. laevis has become the predominant amphibian research model. Given that their allotetraploid genome has complicated the generation of gene knockouts, strategies need to be established for efficient mutagenesis of multiple homeologs to evaluate gene function. Here we describe a protocol to use CRISPR-Cas9-mediated genome editing to target either single alleles or multiple alloalleles in F
0 X. laevis embryos. A single-guide RNA (sgRNA) is designed to target a specific DNA sequence encoding a critical protein domain. To mutagenize a gene with two alloalleles, the sgRNA is designed against a sequence that is common to both homeologs. This sgRNA, along with the Cas9 protein, is microinjected into the zygote to disrupt the genomic sequences in the whole embryo or into a specific blastomere for tissue-targeted effects. Error-prone repair of CRISPR-Cas9-generated DNA double-strand breaks leads to insertions and deletions creating mosaic gene lesions within the embryos. The genomic DNA isolated from each mosaic F0 embryo is sequenced, and software is applied to assess the nature of the mutations generated and degree of mosaicism. This protocol enables the knockout of genes within the whole embryo or in specific tissues in F0 X. laevis embryos to facilitate the evaluation of resulting phenotypes., (© 2022 Cold Spring Harbor Laboratory Press.)- Published
- 2022
- Full Text
- View/download PDF
20. Transgenic Xenopus laevis Line for In Vivo Labeling of Nephrons within the Kidney.
- Author
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Corkins ME, Hanania HL, Krneta-Stankic V, DeLay BD, Pearl EJ, Lee M, Ji H, Davidson AJ, Horb ME, and Miller RK
- Abstract
Xenopus laevis embryos are an established model for studying kidney development. The nephron structure and genetic pathways that regulate nephrogenesis are conserved between Xenopus and humans, allowing for the study of human disease-causing genes. Xenopus embryos are also amenable to large-scale screening, but studies of kidney disease-related genes have been impeded because assessment of kidney development has largely been limited to examining fixed embryos. To overcome this problem, we have generated a transgenic line that labels the kidney. We characterize this cdh17 :eGFP line, showing green fluorescent protein (GFP) expression in the pronephric and mesonephric kidneys and colocalization with known kidney markers. We also demonstrate the feasibility of live imaging of embryonic kidney development and the use of cdh17 :eGFP as a kidney marker for secretion assays. Additionally, we develop a new methodology to isolate and identify kidney cells for primary culture. We also use morpholino knockdown of essential kidney development genes to establish that GFP expression enables observation of phenotypes, previously only described in fixed embryos. Taken together, this transgenic line will enable primary kidney cell culture and live imaging of pronephric and mesonephric kidney development. It will also provide a simple means for high-throughput screening of putative human kidney disease-causing genes., Competing Interests: The authors declare no conflict of interest. The funding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.
- Published
- 2018
- Full Text
- View/download PDF
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