Maternal age related changes in transcriptome of bovine granulosa cells of the dominant follicle at the time of selection
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ABSTRACT:
ORGANISM(S): Bos taurus
SUBMITTER: Muhammad Khan
PROVIDER: E-GEOD-52480 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:The objective of the study was to determine how maternal age influences the transcriptome of the dominant follicle during the preovulatory period. We tested the hypotheses that delayed ovulation in aged cows is associated with 1) altered gene expression of granulosa cells of preovulatory follicles (24 h after LH treatment) and 2) decreased synthesis of progesterone by granulosa cells of the preovulatory follicle. Granulosa cells of preovulatory follicles obtained 24 h after LH treatment from aged Hereford cows (19.0 ±2.5 years; n=3) were compared to those from young cows (9.0 ± 0.6 years; n=3) using bovine specific microarrays (EmbryoGENE-EMBV3; GPL13226). Results were confirmed by RT-qPCR. A total of 1340 genes or gene isoforms were expressed differentially (â¥2-fold change; p ⤠0.05) in aged cows vs. young cows (daughters of aged cows). In conclusion, transcriptome analysis of granulosa cells from aged cows revealed a delayed or suboptimal response to the preovulatory LH stimulus, represented by delayed cellular differentiation, luteinization and progesterone synthesis. Granulosa cells of the dominant preovulatory follicle 24 h after LH treatment were compared between aged vs.young cows. Three biological replicates (each composed of one aged and one young cow). 3 Three technical replicate (dye swap). One biological or technical replicate per array.
2016-06-16 | E-GEOD-54138 | biostudies-arrayexpress
Project description:The fertility of dairy cows is challenged during early lactation and better nutritional strategies need to be developed to address this issue. Combined supplementation of folic acid and vitamin B12 improves energy metabolism in the dairy cow during early lactation. Therefore, the present study was undertaken to explore the effects of this supplement on gene expression in granulosa cells from the dominant follicle during the postpartum period. Multiparous Holstein cows received weekly intramuscular injection of 320 mg folic acid and 10 mg vitamin B12 (treated group) beginning 24 (SD 4) d before calving until 56 d after calving, whereas the control group received saline. The urea plasma concentration was significantly decreased during the pre-calving period, and the concentration of both folate and vitamin B12 were increased in treated animals. Milk production and dry matter intake were not significantly different between the two groups. Plasma concentrations of folates and vitamin B12 were increased in vitamin-treated animals. Daily dry matter intake was not significantly different between the 2 groups before (13.5 kg SE 0.5) and after (23.6 kg SE 0.9) calving. Average energy-corrected milk tended to be greater in vitamin-treated cows, 39.7 (SE 1.4) and 38.1 (SE 1.3) kg/d for treated and control cows, respectively. After calving, average plasma concentration of BHBA tended to be lower in cows injected with the vitamin supplement, 0.47 (SE 0.04) vs. 0.55 (SE 0.03) for treated and control cows, respectively. The ovarian follicle ? 12 mm in diameter was collected by ovarian pick-up after estrus synchronization. Recovered follicular fluid volumes were greater in the vitamin-treated group. A microarray platform was used to investigate the impact of treatment on gene expression of granulosa cells. Lower expression of genes involved in the cell cycle and higher expression of genes associated with granulosa cell differentiation prior to ovulation were observed. Selected candidate genes were analyzed by reverse transcription quantitative polymerase chain reaction. Although the effects of intramuscular injections of folic acid and vitamin B12 on lactational performance and metabolic status of animals were limited, Ingenuity Pathway Analysis of gene expression in granulosa cells suggests a stimulation of cell differentiation in vitamin-treated cows, which may be the result of an increase in LH secretion. Two conditions experiment (Control and Treated). Granulosa cells from the 66h post second PGF2alpha injection. Biological replicates: 3 from each time point. Two technical replicates for each comparison (dye-swap).
2015-09-15 | E-GEOD-72038 | biostudies-arrayexpress
Project description:The objective was to determine age-associated changes in the transcriptome of granulosa cells recovered from the dominant follicle at the time of selection. Granulosa cells were collected from the dominant follicle of aged and young cows after ovariectomy (15 ±1.5 years, n=3 and 6 ± 1.1 years, n=3), or ultrasound-guided follicle aspiration (16±2.1 years, n=4 and 7±1.2 years, 6 ± 1.1 years, n=4) 3-days after ovulation (Day 0). Messenger-RNA was extracted, amplified, labeled with florescent dyes, and hybridized with bovine-specific microarrays (GEO accession # GPL13226). Target intensities were analyzed to determine differential gene expression in granulosa cells from aged vs. young cows. A total 169 genes were differentially expressed (≥ 2 fold-change; P≤0.05) between age groups. In conclusion, granulosa cells collected at the time of selection of the dominant follicle exhibited age-related changes in the transcriptome that may explain follicle-associated loss of oocyte competence in aged cows.
2016-03-31 | GSE52480 | GEO
Project description:The objective of the study was to determine how maternal age influences the transcriptome of the dominant follicle during the preovulatory period. We tested the hypotheses that delayed ovulation in aged cows is associated with 1) altered gene expression of granulosa cells of preovulatory follicles (24 h after LH treatment) and 2) decreased synthesis of progesterone by granulosa cells of the preovulatory follicle. Granulosa cells of preovulatory follicles obtained 24 h after LH treatment from aged Hereford cows (19.0 ±2.5 years; n=3) were compared to those from young cows (9.0 ± 0.6 years; n=3) using bovine specific microarrays (EmbryoGENE-EMBV3; GPL13226). Results were confirmed by RT-qPCR. A total of 1340 genes or gene isoforms were expressed differentially (≥2-fold change; p ≤ 0.05) in aged cows vs. young cows (daughters of aged cows). In conclusion, transcriptome analysis of granulosa cells from aged cows revealed a delayed or suboptimal response to the preovulatory LH stimulus, represented by delayed cellular differentiation, luteinization and progesterone synthesis.
2016-06-16 | GSE54138 | GEO
Project description:In mammals, ovarian folliculogenesis leading to the ovulation of completely mature oocytes is a long and complex process that is regulated at different levels. The mechanisms that underlie the selection of one or several dominant follicles as well as the regulation of the number of ovulating follicles are largely unknown. In this project, we proposed to study the genetic determinism that underlies the difference of ovulation rate between species (cattle and pigs), by studying two processes that exist in ovary: follicular development and follicular atresia. Towards this purpose, we made a comparative transcriptomics study on granulosa cells. Pig and cattle comparison was achieved by a transcriptome analysis with a 9K nylon pig microarray (GPL3729) on granulosa cells from either small healthy antral follicles (SHF), small atretic follicles (SAF) or large healthy antral follicles (LHF). The images were quantified using AGscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Transcriptomic analysis on pig and cattle evidenced 997 differentially expressed genes (FDR1%) between the three follicle classes and/or the two species. This research project which implicated three laboratories from INRA: "Laboratoire de Genetique Cellulaire" (UMR444-LGC) , "Station d'Amelioration Genetique des Animaux" (UR 631-SAGA) and "Physiologie des Comportements et de la reproduction" (UMR 85-PRC) benefited from both European funding through SABRE project and French ANR funding through GenOvul project. Keywords: transcriptome analysis, pig, cattle, ovary, folliculogenesis, gene expression, cDNA microarray The data were obtained from 34 RNA samples: 10 small healthy follicles samples (6 for sows and 4 for cows), 13 small atretic follicles samples (7 for sows and 6 for cows) and 11 samples for large healthy follicles (6 for sows and 5 for cows). They were hybridized on a 9K pig nylon microarray (GPL3729).
2012-12-31 | E-GEOD-24221 | biostudies-arrayexpress
Project description:In mammals, ovarian folliculogenesis leading to the ovulation of completely mature oocytes is a long and complex process that is regulated at different levels. The mechanisms that underlie the selection of one or several dominant follicles as well as the regulation of the number of ovulating follicles are largely unknown. In this project, we proposed to study the genetic determinism that underlies the difference of ovulation rate between species (cattle and pigs). Towards this purpose, we made a comparative transcriptomics study on granulosa cells. Pig and cattle comparison was achieved by a transcriptome analysis with a 9K nylon pig microarray (GPL3729) on granulosa cells from either small healthy antral follicles (SHF) or large healthy antral follicles (LHF). The images were quantified using AGscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Transcriptomic analysis on pig and cattle evidenced 252 differentially expressed genes (FDR5%) between the two follicle classes and/or the two species. This research project which implicated three laboratories from INRA: M-BM-+ Laboratoire de GM-CM-)nM-CM-)tique Cellulaire M-BM-; (UMR444-LGC) , M-BM-+ Station dM-bM-^@M-^YAmM-CM-)lioration GM-CM-)nM-CM-)tique des Animaux M-BM-; (UR 631-SAGA) and M-BM-+ Physiologie des Comportements et de la reproduction M-BM-; (UMR 85-PRC) benefited from both European funding through SABRE project and French ANR funding through GenOvul project. Keywords: transcriptome analysis, pig, cattle, ovary, folliculogenesis, gene expression, cDNA microarray The data were obtained from 21 RNA samples: 10 small healthy follicles samples (6 for sows and 4 for cows) and 11 samples for large healthy follicles (6 for sows and 5 for cows). They were hybridized on a 9K pig nylon microarray (GPL3729).
2012-12-31 | E-GEOD-23335 | biostudies-arrayexpress
Project description:Cows in Negative Energy Balance (NEB) may preferentially divert nutrients away from reproduction, thereby experiencing a period of anovulatory anestrus, delayed ovulation of large follicles and a condition of impaired fertility. To better understand the changes occurring in these large follicles as a function of time post-partum granulosa cells of preovulatory follicles have been collected at different times: 30, 60, 90 and 120 days after calving . An analysis of the transcriptome was performed using a global bovine oligo-array microarray to map the differences in genes expression and cellular functions that occur in the follicular microenvironment during the progressive recovery from NEB condition in dairy cow Four time points experiment: 30, 60, 90 and 120 days. Granulosa cells from the 30, 60 and 90 days compare to the 120 days (reference). Biological replicates: 3 from each time point. One replicate per array.
2013-12-31 | E-GEOD-50854 | biostudies-arrayexpress
Project description:Granulosa cells originating from three different phases of antral follicle growth were compared: growing (G), plateau (P) and atresia (A), as categorized by flow cytometry profiles of DNA. The growing and atretic conditions were each hybridized against the plateau condition as a reference in order to understand the specific biological mechanisms modulated in this class of follicles. Three-condition experiment, Plateau, Growing vs Atretic granulosa cells. Biological replicates: 4 . Dye-swap experiment.
2014-06-02 | E-GEOD-56145 | biostudies-arrayexpress
Project description:Cellular mechanisms that contribute to low estradiol concentrations produced by the preovulatory ovarian follicle in cattle with a compromised metabolic status (such as lactatino) are largely unknown. To gain insight into the main metabolic mechanisms affecting preovulatory follicle function RNAseq profiling was conducted on non-lactating Holstein-Friesian heifers (n=16) and lactating Holstein-Friesian cows (n=17) at three stages of preovulatory follicle development: A) newly selected dominant follicle in the luteal phase (Selection); B) follicular phase before the LH surge (Differentiation) and C) pre-ovulatory phase after the LH surge (Luteinization). Based on a combination of RNA sequencing, ingenuity pathway analysis and Q-RT-PCR validation several important molecular markers involved in steroid biosynthesis, such as the expression of steroidogenic acute regulatory protein (STAR) within developing dominant follicles, were identified to be affected (downregulated) by the catabolic state. We propose that the adverse metabolic environment caused by lactation decreases preovulatory follicle function by affecting cholesterol transport into the mitochondria to initiate steroidogenesis. Granulosa and Theca samples from the dominant follicle were taken from cows and heifers at stages: selection, differentiation and luteinization.
2012-06-15 | E-GEOD-34317 | biostudies-arrayexpress
Project description:In mammals, ovarian folliculogenesis leading to the ovulation of completely mature oocytes is a long and complex process that is regulated at different levels. The mechanisms that underlie the selection of one or several dominant follicles as well as the regulation of the number of ovulating follicles are largely unknown. Atresia is a phenomenon which affects the majority of developing follicles. In this project, we proposed to study the gene regulation of small antral follicles that are either healthy or undergoing atresia in pigs). Towards this purpose, we made a comparative transcriptomics study on granulosa cells, using a 9K nylon pig microarray (GPL3729) on granulosa cells from either small healthy antral follicles (SHF) or small antretic follicles (SAF). The images were quantified using AGscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Transcriptomic analysis evidenced 1682 (912) differentially expressed genes with a 5% (1%) FDR between the two follicle classes. This research project which implicated three laboratories from INRA: "Laboratoire de Genetique Cellulaire" (UMR444-LGC), "Station d'Amelioration Genetique des Animaux" (UR 631-SAGA) and "Physiologie des Comportements et de la reproduction" (UMR 85-PRC) benefited from both European funding through SABRE project and French ANR funding through GenOvul project. Keywords: transcriptome analysis, pig, cattle, ovary, folliculogenesis, gene expression, cDNA microarray The data were obtained from 13 RNA samples: 6 small healthy follicles samples and 7 small atretic follicles. They were hybridized on a 9K pig nylon microarray (GPL3729).
2012-12-31 | E-GEOD-24273 | biostudies-arrayexpress