Project description:A large body of evidence suggests that the development and maintenance of corpus luteum (CL) in primates requires the action of LH. Earlier, using CET-induced luteolysis model, we demonstrated changes in luteal transcriptome suggesting nuclear actions of LH in the primate CL. To further demonstrate the role of LH in maintenance of primate CL, replacement studies were carried out and it was observed that administration of a single injection of rhLH was sufficient to restore the progesterone to pre-CET treatment levels and prevent the CET-induced luteolysis. To elucidate the molecular mechanisms underlying the rescue of CL function, we used LH-replacement model to study immediate early changes in gene expression at a global level (Affymetrix oligonucleotide microarray) following LH replacement in CET-treated monkeys and to evaluate if the changes in gene expression mediated by LH-withdrawal can be reversed by LH replacement. Results demonstrated up-and down-regulation of various genes following LH replacement and suggested that LH-withdrawal induced changes in gene expression are reversible at least for some genes. Keywords: CL, CET, rhLH
Project description:Corpus luteum (CL) is an ephemeral gland whose main function is to secrete progesterone required for the establishment and maintenance of pregnancy. In non-fertile cycles, primate CL has a finite life span of 14-16 days, following which it undergoes regression. Although it has been suggested long time back that PGF2alpha of intra-ovarian or intra-luteal origin acts as a physiological luteolysin in primates, the mechanisms by which PGF2alpha mediates its luteolytic actions are poorly understood. Earlier, we standardized an induced luteolysis model, where 3 injections of PGF2alpha to female bonnet monkeys on day 10 of luteal phase led to luteolysis. To delineate the mechanism of this PGF2alpha-induced luteolysis, we tried to study the global changes in gene expression following PGF2alpha treatment using Affymetrix microarray analysis of PGF2alpha and VEH treated CL and results suggested that PGF2alpha exerts its luteolytic effects by altering gene expression. Keywords: CL, PGF2alpha, VEH, gene expression
Project description:Though uncommon, adoption of orphaned infants has been observed in both wild and captive non-human primates. In two groups of wild bonnet macaques (Macaca radiata), we observed five instances of infants being cared for after they lost their mothers at a pre-weaning age (< 6 months). Orphaned infants had one or more caregivers (juvenile, subadult, and adult female or male) involved in carrying, grooming, hugging, and protecting them. Adoption did not appear to be related to the age/sex class of the infant, or directly to the mother's rank. Although the dominance rank of the mother of an orphaned infant did not have a direct effect on orphan survivorship, it determined the number of caregivers available to the orphaned infant, and infant survivorship was positively related to the number of caregivers of the orphaned infant. Thus, survivorship was likely a function of the mother's sociality. Two other infants born to high-ranking mothers were also adopted by more individuals and survived longer than the infants of low-ranking mothers.
Project description:This study was designed to provide a genome-wide analysis of the effects of luteinizing hormone (LH) ablation/replacement versus steroid ablation/replacement on gene expression in the developed corpus luteum (CL) in primates during the menstrual cycle. Naturally cycling, female rhesus monkeys were left untreated (Control; n = 4) or received one of the following treatments for three days beginning on Day 9 of the luteal phase: daily injection of the gonadotropin-releasing hormone (GnRH) antagonist (Antide; n = 5), Antide + recombinant human LH (A+LH; n = 4), Antide + LH + the 3b-HSD antagonist Trilostane (A+LH+TRL; n = 4), and Antide + LH + TRL + progesterone replacement with a synthetic progestin R5020 (A+LH+TRL+ R5020; n = 5). On Day 12 of the luteal phase, CL were removed and samples of RNA from individual CL were fluorescently labeled and hybridized to Affymetrix™ rhesus macaque total genome microarrays. The greatest number of altered transcripts was associated with the ablation/replacement of LH, while ablation/replacement of progestin affected fewer transcripts. Replacement of LH during Antide treatment restored expression of most transcripts to control levels. Real-time PCR validation of a subset of transcripts revealed that most expression patterns were similar between microarray and real-time PCR. Analysis of protein levels were subsequently determined for 2 of the transcripts differentially expressed by real-time PCR. This is the first genome-wide analysis of LH and steroid regulation of gene transcription in the developed primate CL. Further analysis of novel transcripts identified in this data set can clarify the relative role for LH and steroids in CL maintenance and luteolysis. Keywords: LH/steroid ablation/replacement in primate mid-late luteal phase corpora lutea 22 samples from Rhesus Macaque corpus luteum hybridized to individual Rhesus Affymentrix Gene Chip Arrays. 5 treatment groups, with at least 4 replicates per treatment.
Project description:To explore chorionic gonadotropin (CG)-regulated gene expression in the primate corpus luteum (CL), adult female rhesus macaques were treated with a model of simulated early pregnancy (SEP). Total RNA was isolated from individual CL and hybridized to AffymetrixM-bM-^DM-" GeneChip Rhesus Macaque Genome Arrays The level of 1192 transcripts changed expression > 2-fold (one-way ANOVA, FDR correction; P<0.05) during SEP when compared to Day 10 untreated controls, and the majority of changes occurred between Days 10 and 12 of SEP. To compare transcript levels between SEP rescued and regressing CL, previously banked rhesus GeneChip array data from the mid- to late and very late luteal phase were analyzed with time-matched intervals in SEP. Comparing RMA-normalized transcripts from the natural cycle with those from luteal rescue revealed 7677 transcripts changing in expression pattern >2 fold (one-way ANOVA, FDR correction; P<0.05) between the two groups. Clustering of samples revealed that the SEP samples possessed the most related transcript expression profiles. Regressed CL (days 18-19, around menses) were the most unlike all other CL. The most affected KEGG pathway was Steroid Biosynthesis, and most significantly absent pathways following SEP treatment includes groups of genes whose products promote cell-death. By further comparing the genome-wide changes in luteal gene expression during rescue in SEP, with those in CL during luteolysis in the natural menstrual cycle, it is possible to identify key regulatory pathways promoting fertility. Simulated early pregnancy (SEP) treatment was begun on day 9 as Duffy and Stouffer (1997) by treatment of females with recombinant human chorionic gonadotropin (hCG; NovarelM-bM-^DM-", Ferring Pharmaceuticals Inc. Parsippany, NJ, USA) in increasing dosages (15, 30,45,90,180,360,720,1440, and 2880 IU) twice daily by intramuscular injection. CL were collected by laparotomy on days 10, 12, 15, and 18, representing 1, 3, 6 and 9 days of hCG treatment (n=4 CL/day). Additionally, luteal day 10 untreated CL were collected to serve as baseline controls for SEP CL. All CL were dissected away from luteal tissue, sectioned, and snap-frozen in liquid nitrogen and stored at -80M-BM-0C until RNA and protein isolation by TRIzolM-BM-. extraction (Invitrogen, Carlsbad, CA, USA) according to manufacturerM-bM-^@M-^Ys protocols.