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7. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei

Author

Wakayama, T., Perry, A.C.f., Zuccotti, M., Johnson, K.R., and Yanagimachi, R.

Subjects
Oocytes -- Research, Mammals -- Research, Embryo transplantation -- Observations, Environmental issues, Science and technology, Zoology and wildlife conservation
Abstract

The factors involved in embryonic development by the introduction of nuclei from somatic cells taken from adult mice into enucleated mouse oocytes, are investigated. Some of the enucleated oocytes, receiving Sertoli or neuronal nuclei, implanted after transfer although none developed beyond 8.5 days, while most of those receiving cumulus nuclei developed in vitro. Many implanted after transfer and a significant proportion developed to term.

Published
1998

9. Recovering Seveso

Author

Garagna, Silvia, Rubini, P. G., Redi, C. A., Zuccotti, M., Meriggi, A., Fanelli, R., and Facchetti, S.

Published
1999

13. Embryonic stem cell differentiation studied by FT-IR spectroscopy

Author

Ami, D, Neri, T, Natalello, A, Mereghetti, P, Doglia, S, Zanoni, M, Zuccotti, M, Garagna, S, Redi, C, AMI, DILETTA, NATALELLO, ANTONINO, Redi, CA, DOGLIA, SILVIA MARIA, Ami, D, Neri, T, Natalello, A, Mereghetti, P, Doglia, S, Zanoni, M, Zuccotti, M, Garagna, S, Redi, C, AMI, DILETTA, NATALELLO, ANTONINO, Redi, CA, and DOGLIA, SILVIA MARIA

Abstract

We propose, here, an FT-IR method to monitor the spontaneous differentiation of murine embryonic stem (ES) cells in their early development. Principal component analysis and subsequent linear discriminant analysis enabled us to segregate stem cell spectra into separate clusters corresponding to different differentiation times - and to identify the most significant spectral changes during differentiation. Between days 4 to 7 of differentiation, these spectral changes in the protein amide I band (1700-1600 cm(-1)) and in the nucleic acid absorption region (1050-850 cm(-1)) indicated that mRNA translation was taking place and that specific proteins were produced, reflecting the appearance of a new phenotype. The DNA/ RNA hybrid bands (954 cm(-1) and 899 cm(-1)) were also observed, suggesting that the transcriptional switch of the genome started at this stage of differentiation, As confirmed by cytochemical assays, the FT-IR approach presented here allows to detect at molecular level the biological events of ES cell differentiation as they take place and to monitor in a rapid way the temporal evolution of the ES cell culture. (C) 2007 Elsevier B.V. All rights reserved.

Published
2008

16. ITALIAN REGISTER FOR HIV-1 INFECTION IN CHILDREN - REPORT UP TO 30-3-1990 (1422 CHILDREN ENROLLED)

Author

Coordinatori: DE MARTINO, Registro Italiano per Infezione da HIV in p. e. d. i. a. t. r. i. a., Tovo, M, Collaboratori: Galli, L, Gabiano, C, Partecipanti:, Caramia, G, Mattia, De, Rizzi, D, Zaniboni, M, Masi, Mg, Trombacco, M, Dallacasa, Mg, Salvioli, P, Brusa, Gp, Quarta, S, Duse, G, Bresciani, M, Cao, E, Corrias, A, Belloni, A, Ibba, M, Spada, P, Zannino, A, Digregorio, L, Sciotto, F, Pignata, A, Bezzi, C, Vierucci, T, Salvi, A, Cecchi, G, Terragna, Mt, Ferrazin, A, Ciravegna, A, Boeri, Bw, Jannuzzi, E, Forni, C, Barbanera, Gl, Meo, M, Plebani, A, Schoeller, A, Principi, Mc, Giovannini, N, Zuccotti, M, Simoni, Gv, Stucchi, L, Ferraris, C, Rancillio, G, Mazzoni, L, Altobelli, Pl, Gambaretto, R, Romano, G, Cutillo, G, Guarino, S, Colomba, A, Bassanetti, A, Zacchello, F, Delia, F, Giaquinto, R, Derossi, C, A, and Consolini, Rita

Published
1991

29. Stem Cells.

Author

Redi, C., Monti, M., Merico, V., Neri, T., Zanoni, M., Zuccotti, M., and Garagna, S.

Published
2007
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35. Genome size and constitutive heterochromatin in Hylobates muelleriand Symphalangus syndactylusand in their viable hybrid

Author

Pellicciari, C., Formenti, D., Zuccotti, M., Stanyon, R., and Romanini, Manfredi

Abstract

Genome size was measured as the amount of Feul-gen-stained DNA in six species of the family Hylobatidae and in a hybrid of the gibbon (Hylobates muelleri) and siamang (Symphalangus syndactylus). The family, on the whole, exhibits a wider range of genome sizes than pongids; in particular, the siamang has about 15% more DNA than the 44-chromosome Hylobates species of the “lar” group. Quantitative analysis of C-heterochromatin in hybrid metaphases showed that the difference in genome size of the parental species correlates with the amount of C-band-positive material. Hylobatids are the only group of primates in which karyotype diversification has taken place with a massive quantitative change in constitutive heterochromatin.

Published
1988
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36. Genome composition in Venezuelan spiny-rats of the genus Proechimys(Rodentia, Echimyidae). I. Genome size, C-heterochromatin and repetitive DNAs in situ hybridization patterns

Author

Garagna, S., Pérez-Zapata, A., Zuccotti, M., Mascheretti, S., Marziliano, N., Redi, C.A., Aguilera, M., and Capanna, E.

Abstract

The genome sizes of the Venezuelan spiny-rats Proechimys guairae guairae (2n = 48) and P. trinitatis (2n = 62) were evaluated and proved to be 12.5 ± 0.5pg and 12.6 ± 0.3pg respectively, the highest so far recorded among mammals; also the C-heterochromatin (32.7 %, Coefficient of Variation [CV] 3.8 and 35.8%, CV4.4) and GC (44.2%, CV2.7 and 43.6%, CV 2.9) contents are very high. Highly repetitive (rep) DNAs were isolated from restriction enzyme digested genomic DNAs of both species. The intra- and inter-specific chromosomal allocations of these rep DNAs were analyzed by direct and cross-hybridizations. Results show that the two genomes harbour several rep DNA families which show both species-specificity and interspecific relatedness in their in situ hybridization patterns. The rep DNA families show an equilocal distribution at both the pericentromeric areas of all chromosomes and in the whole arms of two pairs of the uniarmed group, suggesting co-evolution of the rep DNAs. P. g. guairae BamHI digested DNA, when cloned and sequenced, proved to consist of a long “composite” unit (1,239 bp) containing two copies of each of 75-bp and 110-bp internal subrepeats. Karyotype restructuring between P. g. guairae and P. trinitatis, mainly due to Robertsonian changes, was accompanied by slight intra- and intergenomic movements of the putative satellite DNA families within stable genome sizes and C-heterochromatin contents. We discuss the findings obtained in Proechimys in the light of those regarding the kangaroo rat, the pocket gopher and the house mouse; they support the idea that karyotype restructuring could be the expression of molecular driven events of rep DNA amplification and homogenisation through non-homologous chromosomes.

Published
1997
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37. Karyology and molecular biology

Author

Angelici, M. C., Gramiccia, M., Gradoni, L., Angelici, M. C., Ponzi, M., Birago, C., Battaglia, P. A., Cremisi, F., Vignali, E., Rilji, F., Batistoni, E., Androhico, F., Barsacchi-Pilone, G., Capriglione, T., Olmo, E., Cau, A., Deiana, A. M., Salvadori, S., Laudani, U., Gallent, L., Canovai, R., Esposito, A., Stanyon, H., Gianguzza, M., Dolcemascolo, G., Lunadei, M., Marco, A. del, Valentino, F., Rocchi, A., Manicardi, G. C., Garagna, S., Redi, C. A., Bertolani, R., Zuccotti, M., Miceli, C., La Terza, A., Zhang, S., Orias, E., Motta, C. M., Filosa, S., Feola, S., Fiorentino, S., Anoreuccetti, P., Odierna, G., Cobror, O., Olmo, E., Morescalchi, A., Olmo, E., Odierna, G., Cobror, O., Capriglione, T., Pala, M., Vacca, R. A., Casu, S., Ragghianti, M., Bucci, S., Mancino, G., Lacroix, J. C., Rocchi, A., Lanza, V., Castro, M. di, Rubini, M., Goldoni, D., Lunghi, R., Fontana, F., Salvini, M., Barone, E., Giannessi, M., Nobili, R., Stingo, V., Castaldo, G., Improta, R., Stingo, V., Ciampa, C., and Rocco, L.

Published
1986
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38. Population biology

Author

Andreone, F., Crovella, S., Arduino, P., Bullini, L., Badino, G., Celebrano, G., Peyrot, A., Usai, P., Belfiore, C., Boato, A., Rodino, E., Bolzern, A. M., Cantalles, S., Leonardi, M. Cigada, Caccone, A., Cesaroni, D., Allegrucci, G., Lucarelli, M., Sbordoni, V., Pecora, P., Cicolani, B., Cianchi, R., Corti, M., Ciabatti, C. M., Capanna, E., Deri, P., Dim, F., Giubbilini, P., Fava, G., Fusari, C., Gasperi, G., Malacrida, A., Milani, R., Gianni, A., Bracchi, P., Dini, F., Lanfranco, M., Sella, G., Malacarne, G., Lodi, E., Mancino, G., Bucci, S., Ragghianti, M., Mensi, P., Lattes, A., Salvidio, S., Balletto, E., Moretto, E., Girolami, V., Peruffo, A. Dal Belin, Turchetto, M., Nevo, E., Corti, M., Heth, G., Beiles, A., Simson, S., Nigro, L., Pagani, M., Ricci, C., Zullini, A., Paolucci, M., Botte, V., Rovati, C., Rubini, P. G., Franco, M. G., Vecchi, M., Urbanelli, S., Villani, F., Zuccotti, M., Garagna, S., Redi, C. A., Capanna, E., and Nevo, E.

Published
1986
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39. Spermatogenesis in heterozygotes for Robertsonian chromosomal rearrangements from natural populations of the common shrew, Sorex araneus

Author

Garagna, S., Zuccotti, M., Searle, J. B., Redi, C. A., and Wilkinson, P. J.

Abstract

Summary.Twenty-two adult male common shrews were collected from 5 sites in the vicinity of Oxford (UK) close to the zone of hybridization between two karyotypic races. The shrews were subdivided into 3 karyotypic categories: homozygotes, simple Robertsonian heterozygotes (which form one or more trivalents at prophase I of meiosis) and complex Robertsonian heterozygotes (which form a quadrivalent). The ratio of primary spermatocytes to round spermatids was determined from transverse sections of seminiferous tubules, to provide an indication of germ cell death. In no individual was there severe germ cells loss. Homozygotes had the highest mean spermatocyte: spermatid ratio and complex heterozygotes the lowest, but there was substantial individual variation and the differences were not significant. Complex heterozygotes also had a higher proportion of defective seminiferous tubules and lower testis weights than did other categories and it is reasonable to propose that, as a population, complex heterozygotes have reduced fitness relative to other categories on the basis of spermatogenic performance. However, there is no evidence from studies of spermatogenesis that simple Robertsonian heterozygotes are less fit than homozygotes.Keywords:common shrew; Robertsonian rearrangements; spermatogenesis; Robertsonian heterozygotes

Published
1989
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40. Karyology and molecular biology

Author

Angelici, M. C., primary, Gramiccia, M., additional, Gradoni, L., additional, Angelici, M. C., additional, Ponzi, M., additional, Birago, C., additional, Battaglia, P. A., additional, Cremisi, F., additional, Vignali, E., additional, Rilji, F., additional, Batistoni, E., additional, Androhico, F., additional, Barsacchi‐Pilone, G., additional, Capriglione, T., additional, Olmo, E., additional, Cau, A., additional, Deiana, A. M., additional, Salvadori, S., additional, Laudani, U., additional, Gallent, L., additional, Canovai, R., additional, Esposito, A., additional, Stanyon, H., additional, Gianguzza, M., additional, Dolcemascolo, G., additional, Lunadei, M., additional, del Marco, A., additional, Valentino, F., additional, Rocchi, A., additional, Manicardi, G. C., additional, Garagna, S., additional, Redi, C. A., additional, Bertolani, R., additional, Zuccotti, M., additional, Miceli, C., additional, La Terza, A., additional, Zhang, S., additional, Orias, E., additional, Motta, C. M., additional, Filosa, S., additional, Feola, S., additional, Fiorentino, S., additional, Anoreuccetti, P., additional, Odierna, G., additional, Cobror, O., additional, Morescalchi, A., additional, Pala, M., additional, Vacca, R. A., additional, Casu, S., additional, Ragghianti, M., additional, Bucci, S., additional, Mancino, G., additional, Lacroix, J. C., additional, Lanza, V., additional, di Castro, M., additional, Rubini, M., additional, Goldoni, D., additional, Lunghi, R., additional, Fontana, F., additional, Salvini, M., additional, Barone, E., additional, Giannessi, M., additional, Nobili, R., additional, Stingo, V., additional, Castaldo, G., additional, Improta, R., additional, Ciampa, C., additional, and Rocco, L., additional

Published
1986
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42. Population biology

Author

Andreone, F., primary, Crovella, S., additional, Arduino, P., additional, Bullini, L., additional, Badino, G., additional, Celebrano, G., additional, Peyrot, A., additional, Usai, P., additional, Belfiore, C., additional, Boato, A., additional, Rodino, E., additional, Bolzern, A. M., additional, Cantalles, S., additional, Leonardi, M. Cigada, additional, Caccone, A., additional, Cesaroni, D., additional, Allegrucci, G., additional, Lucarelli, M., additional, Sbordoni, V., additional, Pecora, P., additional, Cicolani, B., additional, Cianchi, R., additional, Corti, M., additional, Ciabatti, C. M., additional, Capanna, E., additional, Deri, P., additional, Dim, F., additional, Giubbilini, P., additional, Fava, G., additional, Fusari, C., additional, Gasperi, G., additional, Malacrida, A., additional, Milani, R., additional, Gianni, A., additional, Bracchi, P., additional, Dini, F., additional, Lanfranco, M., additional, Sella, G., additional, Malacarne, G., additional, Lodi, E., additional, Mancino, G., additional, Bucci, S., additional, Ragghianti, M., additional, Mensi, P., additional, Lattes, A., additional, Salvidio, S., additional, Balletto, E., additional, Moretto, E., additional, Girolami, V., additional, Peruffo, A. Dal Belin, additional, Turchetto, M., additional, Nevo, E., additional, Heth, G., additional, Beiles, A., additional, Simson, S., additional, Nigro, L., additional, Pagani, M., additional, Ricci, C., additional, Zullini, A., additional, Paolucci, M., additional, Botte, V., additional, Rovati, C., additional, Rubini, P. G., additional, Franco, M. G., additional, Vecchi, M., additional, Urbanelli, S., additional, Villani, F., additional, Zuccotti, M., additional, Garagna, S., additional, and Redi, C. A., additional

Published
1986
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43. Addendum

Author

Juberthie, C., primary, Pellicciari, C., additional, Formenti, D., additional, Zuccotti, M., additional, Stanyon, R., additional, and Manfredi‐Romanini, H. G., additional

Published
1986
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44. Transcriptome based identification of mouse cumulus cell markers that predict the developmental competence of their enclosed antral oocytes

Author

Vigone, G., Merico, V., Prigione, A., Mulas, F., Sacchi, L., Gabetta, M., Bellazzi, R., Redi, C.A., Mazzini, G., Adjaye, J., Garagna, S., and Zuccotti, M.

Subjects
Oocyte, Cumulus Cells, fungi, Developmental competence, Mice, Pregnancy, Oocytes, Genetics, Animals, Female, Function and Dysfunction of the Nervous System, Transcriptome, Biomarkers, Cell Nucleolus, Research Article, Oligonucleotide Array Sequence Analysis, Biotechnology
Abstract

BACKGROUND: The cumulus cells (CCs) enveloping antral and ovulated oocytes have been regarded as putative source of non-invasive markers of the oocyte developmental competence. A number of studies have indeed observed a correlation between CCs gene expression, embryo quality, and final pregnancy outcome. Here, we isolated CCs from antral mouse oocytes of known developmental incompetence (NSN-CCs) or competence (SN-CCs) and compared their transcriptomes with the aim of identifying distinct marker transcripts. RESULTS: Global gene expression analysis highlighted that both types of CCs share similar transcriptomes, with the exception of 422 genes, 97.6% of which were down-regulated in NSN-CCs vs. SN-CCs. This transcriptional down-regulation in NSN-CCs was confirmed by qRT-PCR analysis of CC-related genes (Has2, Ptx3, Tnfaip6 and Ptgs2). Only ten of the 422 genes were up-regulated with Amh being the most up-regulated in NSN-CCs, with an average 4-fold higher expression when analysed by qRT-PCR. CONCLUSIONS: The developmental incompetence (NSN) or competence (SN) of antral oocytes can be predicted using transcript markers expressed by their surrounding CCs (i.e., Has2, Ptx3, Tnfaip6, Ptgs2 and Amh). Overall, the regulated nature of the group of genes brought out by whole transcriptome analysis constitutes the molecular signature of CCs associated either with developmentally incompetent or competent oocytes and may represent a valuable resource for developing new molecular tools for the assessment of oocyte quality and to further investigate the complex bi-directional interaction occurring between CCs and oocyte.

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45. Addendum

Author

Juberthie, C., Pellicciari, C., Formenti, D., Zuccotti, M., Stanyon, R., and Manfredi-Romanini, H. G.

Published
1986
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47. A Human Homologue of the Drosophila melanogaster diaphanous Gene Is Disrupted in a Patient with Premature Ovarian Failure: Evidence for Conserved Function in Oogenesis and Implications for Human Sterility

Author

Maurizio Zuccotti, Giuseppe Borsani, Sandro Banfi, Andrea Ballabio, Giulia Arrigo, Orsetta Zuffardi, Christophe Philippe, Daniela Toniolo, Cinzia Sala, Philippe Jonveaux, Silvia Bione, Chiara Manzini, Bione, S, Sala, C, Manzini, C, Arrigo, G, Zuffardi, O, Banfi, S, Borsani, G, Jonveaux, P, Philippe, C, Zuccotti, M, Ballabio, Andrea, Toniolo, D., Banfi, Sandro, and Ballabio, A

Subjects
Ovarian dysgenesis, endocrine system diseases, Messenger, Sequence Homology, Primary Ovarian Insufficiency, Oogenesis, Translocation, Genetic, Pair 12, Drosophila Proteins, Developmental, Genetics(clinical), Genetics (clinical), Diaphanous gene, Genetics, biology, Gene Expression Regulation, Developmental, Chromosome Mapping, Menopause, early, female genital diseases and pregnancy complications, Premature ovarian failure, Amino Acid, Drosophila melanogaster, Female, Infertility, Female, Drosophila Protein, Research Article, Human, X Chromosome, Protein family, Sterility, X-chromosome rearrangements, Turner syndrome, Molecular Sequence Data, Formins, Translocation, Chromosomes, Genetic, Ovarian failure, Drosophilidae, medicine, Amino Acid Sequence, Animals, Carrier Proteins, Chromosomes, Human, Pair 12, Humans, Ovary, RNA, Messenger, Sequence Homology, Amino Acid, Gene, biology.organism_classification, medicine.disease, Gene Expression Regulation, Infertility, RNA
Abstract

SummaryPremature ovarian failure (POF) is a defect of ovarian development and is characterized by primary or secondary amenorrhea, with elevated levels of serum gonadotropins, or by early menopause. The disorder has been attributed to various causes, including rearrangements of a large “critical region” in the long arm of the X chromosome. Here we report identification, in a family with POF, of a gene that is disrupted by a breakpoint. The gene is the human homologue of the Drosophila melanogaster diaphanous gene; mutated alleles of this gene affect spermatogenesis or oogenesis and lead to sterility. The protein (DIA) encoded by the human gene (DIA) is the first human member of the growing FH1/FH2 protein family. Members of this protein family affect cytokinesis and other actin-mediated morphogenetic processes that are required in early steps of development. We propose that the human DIA gene is one of the genes responsible for POF and that it affects the cell divisions that lead to ovarian follicle formation.

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48. Embryonic stem cell differentiation studied by FT-IR spectroscopy

Author

Silvia Garagna, Mario Zanoni, Maurizio Zuccotti, Silvia Maria Doglia, Paolo Mereghetti, Antonino Natalello, Tui Neri, Carlo Alberto Redi, Diletta Ami, Ami, D, Neri, T, Natalello, A, Mereghetti, P, Doglia, S, Zanoni, M, Zuccotti, M, Garagna, S, and Redi, C

Subjects
Cytodifferentiation, Linear discriminant analysis, Cellular differentiation, linear discriminant analysi, Principal component analysis, Cardiomyocyte, Biology, chemistry.chemical_compound, Mice, Spectroscopy, Fourier Transform Infrared, Animals, Myocytes, Cardiac, Molecular Biology, Cell Shape, Cells, Cultured, Embryonic Stem Cells, RNA, Proteins, Fourier transform infrared spectroscopy, Cell Differentiation, Cell Biology, Embryonic stem cell, Phenotype, Amides, Cell biology, chemistry, Cell culture, Multivariate Analysis, Nucleic acid, Stem cell, DNA
Abstract

We propose, here, an FT-IR method to monitor the spontaneous differentiation of murine embryonic stem (ES) cells in their early development. Principal component analysis and subsequent linear discriminant analysis enabled us to segregate stem cell spectra into separate clusters corresponding to different differentiation times - and to identify the most significant spectral changes during differentiation. Between days 4 to 7 of differentiation, these spectral changes in the protein amide I band (1700-1600 cm(-1)) and in the nucleic acid absorption region (1050-850 cm(-1)) indicated that mRNA translation was taking place and that specific proteins were produced, reflecting the appearance of a new phenotype. The DNA/ RNA hybrid bands (954 cm(-1) and 899 cm(-1)) were also observed, suggesting that the transcriptional switch of the genome started at this stage of differentiation, As confirmed by cytochemical assays, the FT-IR approach presented here allows to detect at molecular level the biological events of ES cell differentiation as they take place and to monitor in a rapid way the temporal evolution of the ES cell culture. (C) 2007 Elsevier B.V. All rights reserved.

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49. Cytoplasmic strings in human blastocysts: hypotheses of their role and implications for embryo selection.

Author

Marconetto A, Innocenti F, Saturno G, Taggi M, Chiappetta V, Trio S, De Falco F, Albricci L, Coticchio G, Ahlström A, Fiorentino G, Maggiulli R, Vaiarelli A, Zuccotti M, Rienzi L, and Cimadomo D

Subjects
Humans, Female, Retrospective Studies, Pregnancy, Adult, Cytoplasm metabolism, Aneuploidy, Vitrification, Embryo Transfer methods, Fertilization in Vitro, Embryo Implantation physiology, Blastocyst, Embryo Culture Techniques, Embryonic Development physiology
Abstract

Study Question: What are the implications of the presence cytoplasmic strings (Cyt-S) and their quantity and dynamics for the pre-implantation development of human blastocysts?, Summary Answer: Cyt-S are common in human embryos and are associated with faster blastocyst development, larger expansion, and better morphological quality., What Is Known Already: Cyt-S are dynamic cellular projections connecting inner cell mass and trophectoderm (TE) cells, that can be observed during blastocyst expansion. Their prevalence in human embryos has been estimated to be between 44% and 93%. Data relevant to their clinical implications and role in development are lacking, limited, or controversial., Study Design, Size, Duration: Retrospective study conducted at a single IVF center between May 2013 and November 2014 and involving 124 pre-implantation genetic testing for aneuploidy cycles in a time-lapse incubator with ≥1 blastocyst biopsied and vitrified (N = 370 embryos assessed). These cycles resulted in 87 vitrified-warmed single-euploid blastocyst transfers., Participants/materials, Setting, Methods: ICSI, continuous blastocyst culture (Days 5-7), TE biopsy of fully expanded blastocysts without Day 3 zona pellucida drilling, qPCR to assess uniform full-chromosome aneuploidies, and vitrification were all performed. Only vitrified-warmed euploid single-embryo-transfers were conducted. Blastocyst morphological quality was defined according to Gardner's criteria. The AI-based software CHLOE™ (Fairtility) automatically registered timings from time of starting blastulation (tSB) to biopsy (t-biopsy, i.e. blastocyst full-expansion) as hours-post-insemination (hpi), embryo area (including zona pellucida in µm2), and spontaneous blastocyst collapses. One senior embryologist manually annotated Cyt-S presence, quantity, timings, and type (thick cell-to-cell connections and/or threads). All significant associations were confirmed through regression analyses. All couples', cycles', and embryos' main features were also tested for associations with Cyt-S presence, quantity, and dynamics., Main Results and the Role of Chance: About 94.3% of the patients (N = 117/124) had ≥1 embryo with Cyt-S. Out of a total of 370 blastocysts, 55 degenerated between blastulation and full-expansion (N = 55/370, 14.9%). The degeneration rate among embryos with ≥1 Cyt-S was 10.8% (N = 33/304), significantly lower than that of embryos without Cyt-S (33.3%, N = 22/66, P < 0.01). Of the remaining 315 viable blastocysts analyzed, 86% (N = 271/315; P < 0.01) had ≥1 Cyt-S, on average 3.5 ± 2.1 per embryo ranging 1-13. The first Cyt-S per viable embryo appeared at 115.3 ± 12.5 hpi (85.7-157.7), corresponding to 10.5 ± 5.8 h (0.5-31) after tSB. Overall, we analyzed 937 Cyt-S showing a mean duration of 3.8 ± 2.7 h (0.3-20.9). Cyt-S were mostly threads (N = 508/937, 54.2%) or thick cell-to-cell connections becoming threads (N = 382/937, 40.8%) than thick bridges (N = 47/937, 5.0%). The presence and quantity of Cyt-S were significantly associated with developmentally faster (on average 6-12 h faster) and more expanded (on average 2700 µm2-larger blastocyst's area at t-biopsy) embryos. Also, the presence and duration of Cyt-S were associated with better morphology. Lastly, while euploidy rates were comparable between blastocysts with and without Cyt-S, all euploid blastocysts transferred from the latter group failed to implant (N = 10)., Limitations, Reasons for Caution: Cyt-S presence and dynamics were assessed manually on seven focal planes from video frames recorded every 15 min. The patients included were mostly of advanced maternal age. Only associations could be reported, but no causations/consequences. Lastly, larger datasets are required to better assess Cyt-S associations with clinical outcomes., Wider Implications of the Findings: Cyt-S are common during human blastocyst expansion, suggesting their physiological implication in this process. Their presence, quantity and dynamics mirror embryo viability, and morphological quality, yet their role is still unknown. Future basic science studies are encouraged to finally describe Cyt-S molecular nature and biophysical properties, and Artificial Intelligence tools should aid these studies by incorporating Cyt-S assessment., Study Funding/competing Interest(s): None., Trial Registration Number: N/A., (© The Author(s) 2024. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Published
2024
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50. BPA Exposure Affects Mouse Gastruloids Axial Elongation by Perturbing the Wnt/β-Catenin Pathway.

Author

Rebuzzini P, Rustichelli S, Fassina L, Canobbio I, Zuccotti M, and Garagna S

Subjects
Animals, Mice, Cadherins metabolism, Cadherins genetics, Organoids metabolism, Organoids drug effects, Gene Expression Regulation, Developmental drug effects, Embryonic Development drug effects, Cell Differentiation drug effects, Endocrine Disruptors toxicity, Phenols toxicity, Benzhydryl Compounds toxicity, Wnt Signaling Pathway drug effects, beta Catenin metabolism, beta Catenin genetics
Abstract

Mammalian embryos are very vulnerable to environmental toxicants (ETs) exposure. Bisphenol A (BPA), one of the most diffused ETs, exerts endocrine-disrupting effects through estro-gen-mimicking and hormone-like properties, with detrimental health effects, including on reproduction. However, its impact during the peri-implantation stages is still unclear. This study, using gastruloids as a 3D stem cell-based in vitro model of embryonic development, showed that BPA exposure arrests their axial elongation when present during the Wnt/β-catenin pathway activation period by β-catenin protein reduction. Gastruloid reshaping might have been impeded by the downregulation of Snail, Slug and Twist, known to suppress E-cadherin expression and to activate the N-cadherin gene, and by the low expression of the N-cadherin protein. Also, the lack of gastruloids elongation might be related to altered exit of BPA-exposed cells from the pluripotency condition and their following differentiation. In conclusion, here we show that the inhibition of gastruloids' axial elongation by BPA might be the result of the concomitant Wnt/β-catenin perturbation, reduced N-cadherin expression and Oct4, T/Bra and Cdx2 altered patter expression, which all together concur in the impaired development of mouse gastruloids.

Published
2024
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