Understanding the underlying molecular mechanisms of meiotic arrest in organotypic culture of prepubertal rat testicular tissues
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ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE197241 | GEO | 2022/06/15
REPOSITORIES: GEO
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Project description:Understanding the underlying molecular mechanisms of meiotic arrest in organotypic culture of prepubertal rat testicular tissues
| PRJNA809412 | ENA
Project description:Antineoplastic treatments for cancer and other non-malignant disorders can result in male long-term or permanent infertility by ablating spermatogonial stem cells (SSCs). Preserving the future fertility of prepubertal patients faced with this prospect cannot rely on sperm banking. SSC transplantation using testicular tissue harvested before sterilizing treatment is a promising approach for restoring fertility, but lack of exclusive biomarkers which can unequivocally identify prepubertal SSCs in any primate species limits the therapeutic potential. To address this, we performed single-cell RNA-seq on unselected testis cells from immature baboons and macaques and compared these cells with published data from pre-pubertal human germ cells functionally-defined mouse SSCs. We found discrete groups of human spermatogonia while baboon and rhesus spermatogonia appeared less heterogenous. Cross-species analysis revealed cell types analogous to human SSCs in baboon and rhesus germ cells, but comparison with mouse SSCs revealed significant differences with primate SSCs. Indeed, a core component of a human SSC gene expression signature was conserved in baboon and rhesus spermatogonia and absent from mouse SSCs. These results further resolve the identity of pre-pubertal human SSCs and define novel pathways that could be leveraged for advancing their selection and propagation in vitro.
2023-05-15 | GSE222105 | GEO
Project description:The present study was undertaken to discover molecular markers in bovine cumulus cells predictive of oocyte competence and elucidate their functional significance. Differences in RNA transcript abundance in cumulus cells harvested from oocytes of adult versus prepubertal animals (model of poor oocyte quality) were identified by microarray analysis. Four genes of interest encoding for the lysosomal cysteine proteinases cathepsin B, S, K and Z and displaying greater transcript abundance in cumulus cells surrounding oocytes harvested from prepubertal animals were chosen for further investigation. Greater mRNA abundance for such genes in cumulus cells of prepubertal oocytes was confirmed by real time RT-PCR. Elevated transcript abundance for cathepsins B, S and Z was also observed in cumulus cells surrounding adult metaphase II oocytes that developed to the blastocyst stage at a low percentage following parthenogenetic activation, versus those that developed at a high percentage. Functional significance of cumulus cell cathepsin expression to oocyte competence was confirmed by treatment of cumulus oocyte complexes during in vitro oocyte maturation with a cell permeable cysteine proteinase (cathepsin) inhibitor. Inhibitor treatment decreased apoptotic nuclei in the cumulus layer and enhanced development of parthenogenetically activated and in vitro fertilized adult oocytes to the blastocyst stage. Stimulatory effects of inhibitor treatment during meiotic maturation on subsequent embryonic development were not observed when oocytes were matured in the absence of cumulus cells. Results support a functional role for cumulus cell cathepsins in compromised oocyte competence and suggest that cumulus cell cathepsin mRNA abundance may be predictive of oocyte quality. Keywords: Bovine, microarray, cDNA, cumulus cells Differences in RNA transcript abundance in cumulus cells harvested from oocytes of adult versus prepubertal animals (model of poor oocyte quality) were identified by microarray analysis. Total RNA from pools of cumulus cells (n = 4) collected from adult and prepubertal animals for microarray experiments was amplified. Two color microarray experiments were conducted using a bovine cDNA array containing expressed sequence tags (ESTs) representing approximately 15200 unique genes. Hybridizations were performed on duplicate slides (prepubertal versus adult) and incorporated a dye swap. The total number of slides used is eight.
2010-05-25 | E-GEOD-10819 | biostudies-arrayexpress
Project description:Developmental competences of oocytes derived from prepubertal heifers are lower than those derived from adult counterparts. The objective of this study was to identify a range of genes associated with reduced oocyte competence that are differentially expressed between adult versus prepubertal donors. Microarray experiments were conducted using total RNA isolated from GV and MII stages oocytes collected from adult and prepubertal animals using Affymetrix GeneChip Bovine Genome Array containing 24,072 probe sets representing over 23,000 transcripts. A total of 549 and 333 genes were differentially expressed between prepubertal versus adult bovine MII and GV stages oocytes respectively. Out of these, 312 and 176 genes were up-regulated, while 237 and 157 were down-regulated in prepubertal when compared with adult MII and GV oocytes respectively. Ontological classification of the differentially expressed genes revealed that up-regulated genes in adult oocytes were involved in signal transduction, regulation of transcription DNA-dependent, and transport. Results from the present study indicated that significant number of genes were differentially expressed (>2-fold, p<0.01) between the two groups. Thus the decreased developmental competence of oocytes from prepubertal heifers may be induced due to difference in gene expression abundance as observed in our study. In conclusion, transcript abundance analyses of oocytes using microarray approach have been carried out in bovine and several other species. However, to our knowledge, this is the first study carried out to examine genes expression differential abundance in oocytes derived from perpubertal versus adult Japanese Black Cattle. Bovine 4b PP biological rep1, Bovine 78b PP biological rep2, Bovine 79 PP biological rep3 represents GV stage oocytes derived from Prepubertal (PP) heifer group, while Bovine 74b A biological rep1, Bovine 80b A biological rep2, Bovine 81 A biological rep3 represents GV stage oocytes derived from Adult (A) cow group. Bovine 7 PP biological rep1, Bovine 53 PP biological rep2, Bovine 57 PP biological rep3 represents MII stage oocytes derived from Prepubertal heifer group, while Bovine 59 A biological rep1, Bovine 70 A biological rep2, Bovine 71 A biological rep3 represents MII stage oocytes from Adult cow group. The ovaries of adult cows (Japanese black cattle) were collected from local abattoir while ovaries of prepubertal Japanese black heifers (9 to 12 months old) were collected by spay device at several commercial farms. The collected ovaries of both the adult cows and prepubertal heifers groups were transported to the research laboratory in 0.67% (w/v) NaCl solution containing 100 mg/L kanamycin sulfate (Meiji Seika, Tokyo, Japan). For both groups, cumulus oocyte complexes (COCs) from ovarian follicles 2 to 8M-BM- mm in diameter were aspirated by using an 18 gauge needle (Terumo co, Tokyo, Japan) attached to a 5 ml disposable syringe (Nipro, Osaka, Japan). After collection, the COCs were washed twice with Tyrode-lactate-pyruvate-polyvinylalcohol (Hepes-TLP-PVA) and TCM 199 (Invitrogen, Gibco, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (maturation medium). Only COCs with evenly granulated cytoplasm surrounded by multiple layers of compact cumulus cells were used in all the experiments. The COCs (70 to 80) were placed in 200 M-BM-5L drop of the maturation medium in petri dish (35x10mm, Becton Dickinson Labware, Oxnard, CA, USA) covered with paraffin liquid (Nacalai Tesque Inc, Kyoto, Japan) and cultured at 38.5M-BM-0C in a humidified atmosphere of 5% CO2 in air for 20 to 22 h for maturation.
2012-05-31 | E-GEOD-24477 | biostudies-arrayexpress
Project description:Age of the donor animal has a significant influence on the oocyteM-bM-^@M-^Ys ability to complete maturation and acquire the mRNA and proteins required for normal embryonic development. It is well known that developmental competence is reduced in oocytes derived from juvenile animals when compared to that derived from adults. However, the molecular mechanisms associated with these differences are not well elucidated. The aim of this study was to analyze the transcriptome differences between adult versus prepubertal derived preimplantation embryos and to identify genes associated with oocyte developmental potential. We performed cDNA microarray analysis on populations of preimplantation embryos (8- to 16-cell and Day 7 blastocysts) derived from adult versus prepubertal Japanese Black cattle. Total RNA was extracted and amplified in a linear fashion and then subjected to the microarray hybridization using the Affymetrix GeneChip Bovine Genome Array. The Bovine Array contains 24 072 probe sets representing over 23000 transcripts and 19000 UniGene clusters. Following normalization of the microarray data, analysis revealed differences in gene expression abundance in 8- to 16-cell and blastocysts harvested from adult versus prepubertal heifers. Results indicated that a total of 591 and 490 genes were differentially expressed (M-bM-^IM-%2-fold) in 8- to 16-cell and blastocyst respectively between the two groups (p<0.01). In 8- to 16-cell stage, 373 genes were up-regulated and 218 gens down-regulated in adult when compared to the prepubertal heifer group. In case of blastocyst stage embryos, 242 genes were up-regulated and 248 genes were down-regulated in adult when compared to prepubertal group. The ovaries of adult cows (Japanese black cattle) were collected from local abattoir while ovaries of prepubertal Japanese black heifers (9 to 12 months old) were collected by spay device at several commercial farms. The ovaries were transported to the research laboratory in 0.67% (w/v) NaCl solution containing 100 mg/L kanamycin sulfate. COC were placed in 200 M-BM-5L drop of the maturation medium (TCM 199 + 10% FBS) and cultured at 38.5M-BM-0C in a humidified atmosphere of 5% CO2 in air for 20 to 22 h for in vitro maturation (IVM). After 20 to 22 h IVM, the COC were fertilized in vitro with frozenM-bM-^@M-^Sthawed semen of Japanese Black bull in BO medium. Six hours after IVF, oocytes were denuded of surrounding cumulus cells by gentle pipetting in a solution of 0.1% (w/v) hyaluronidase. The presumptive zygotes were cultured in CR1aa medium supplemented with fatty acid-free BSA (Sigma) for 2 days and cultured in CR1aa medium supplemented with 10% (v/v) fetal bovine serum (FBS) for next 5 days at 38.5 M-BM-0C in 5% CO2, 5% O2 and 90% N2 with high humidity (15 to 20 zygotes per 50 M-BM-5l microdrop). 8- to 16-cell refers to day 3, 70 to 72 hours post insemination( hpi) preimplantation embryo stage containing 8 to 16 cells, while blastocyst refer to pre-hatching day 7 expanded blastocysts. Pools of 8- to 16- cell embryos (50 in each sample) and blastocysts (10 in each sample) from prepubertal and adult groups were used for cDNA microarray.
2012-05-31 | E-GEOD-24596 | biostudies-arrayexpress
Project description:Defining prepubertal primate spermatogonial stem cells to address fertility preservation in prepubertal boys
| PRJNA917768 | ENA
Project description:The in vitro spermatogenesis procedure, once optimized, should be proposed to survival prepubertal patients at high risk for malignant contamination in their testis, for example in case of acute leukaemia. Differentially expressed genes between the in vitro cultured testes and the aged-matched in vivo controls will be explored by a transcriptomic analysis using the RNA-Seq technology (single-read 50 bp, PolyA-enriched) at D0, D4, D16 and D30 of culture using our reference culture medium (one-step culture), and at the same time points after an initial culture step with growth factors (two-step culture). This approach will allow us to identify the signalling pathways or molecular mechanisms that are deregulated in vitro. Moreover, a comparison of transcriptomic analyses to cryopreserved testicular fragments will allow us to identify the potential impact of cryopreservation procedures for fragments after thawing (D0) and explants at the end of culture with our reference culture medium (D30).
2023-01-31 | GSE221333 | GEO
Project description:Developmental competences of oocytes derived from prepubertal heifers are lower than those derived from adult counterparts. The objective of this study was to identify a range of genes associated with reduced oocyte competence that are differentially expressed between adult versus prepubertal donors. Microarray experiments were conducted using total RNA isolated from GV and MII stages oocytes collected from adult and prepubertal animals using Affymetrix GeneChip Bovine Genome Array containing 24,072 probe sets representing over 23,000 transcripts. A total of 549 and 333 genes were differentially expressed between prepubertal versus adult bovine MII and GV stages oocytes respectively. Out of these, 312 and 176 genes were up-regulated, while 237 and 157 were down-regulated in prepubertal when compared with adult MII and GV oocytes respectively. Ontological classification of the differentially expressed genes revealed that up-regulated genes in adult oocytes were involved in signal transduction, regulation of transcription DNA-dependent, and transport. Results from the present study indicated that significant number of genes were differentially expressed (>2-fold, p<0.01) between the two groups. Thus the decreased developmental competence of oocytes from prepubertal heifers may be induced due to difference in gene expression abundance as observed in our study. In conclusion, transcript abundance analyses of oocytes using microarray approach have been carried out in bovine and several other species. However, to our knowledge, this is the first study carried out to examine genes expression differential abundance in oocytes derived from perpubertal versus adult Japanese Black Cattle. Bovine 4b PP biological rep1, Bovine 78b PP biological rep2, Bovine 79 PP biological rep3 represents GV stage oocytes derived from Prepubertal (PP) heifer group, while Bovine 74b A biological rep1, Bovine 80b A biological rep2, Bovine 81 A biological rep3 represents GV stage oocytes derived from Adult (A) cow group. Bovine 7 PP biological rep1, Bovine 53 PP biological rep2, Bovine 57 PP biological rep3 represents MII stage oocytes derived from Prepubertal heifer group, while Bovine 59 A biological rep1, Bovine 70 A biological rep2, Bovine 71 A biological rep3 represents MII stage oocytes from Adult cow group.
2012-06-01 | GSE24477 | GEO
Project description:Full title: Prepubertal Human Spermatogonia and Mouse Gonocytes Share Conserved Gene Expression of Germline Stem Cell Regulatory Molecules Oligonucleotide Microarray Analysis Reveals Novel Genes Expressed in Prepubertal Human Spermatogonia and Mouse Gonocytes
2009-11-07 | GSE18914 | GEO
Project description:Oocyte developmental potential is progressively obtained as females approach puberty. Therefore, oocytes derived from prepubertal females are less developmentally competent, indicated by decreased embryonic development, compared to oocytes derived from adult females. To investigate mechanisms involved in establishing oocyte cytoplasmic maturation and developmental competence, Affymetrix GeneChip microarrays were used. Keywords: oocyte developmental competence, maternal age Porcine oocytes obtained from prepubertal and adult females were collected for RNA extraction and hybridization on Affymetrix microarrays. Oocytes were aspirated from 2 to 6 mm ovarian follicles and matured in vitro. Analysis of the first extruded polar body ensured that all oocytes used in the analyses had completed nuclear maturation.
2009-12-30 | E-GEOD-11590 | biostudies-arrayexpress