Microarray analysis of the primate luteal transcriptome during chorionic gonadotrophin administration simulating early pregnancy

Bishop, Cecily V.; Satterwhite, Shannon; Xu, Liangzhi; Hennebold, Jon D.; Stouffer, Richard L.

doi:10.1093/molehr/gar073

Cited by 18 publications

(29 citation statements)

References 33 publications

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“…Processed microarray image (.CEL) files were analyzed using the web-based GeneSifter® software (Geospiza, Inc., Seattle, WA, USA) and probeset (mRNA) hybridization data were log2-transformed using the Robust Multi-array Average (RMA) algorithm as previously reported (23). All mRNA expression data were interrogated by one-way ANOVA; standard diet (control) versus WSD versus WSD+T (n=3/ group).…”

Section: Methodsmentioning

confidence: 99%

“…Pairwise comparisons were performed on SAF mRNA levels between standard diet controls, WSD, and WSD+T by Welch’s T-test (≥2 fold, P<0.05). KEGG analysis was performed on the list of significantly altered mRNAs similar to previous methods (23). Significant ontologies identified by GeneSifter were refined by eliminating those in which fewer than 5 gene products were associated with the ontology (too narrow classification), and with greater than 100 gene products (too broad) similar to Doniger et al (24).…”

Section: Methodsmentioning

confidence: 99%

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Western-style diet, with and without chronic androgen treatment, alters the number, structure, and function of small antral follicles in ovaries of young adult monkeys

Bishop

et al. 2016

Fertility and Sterility

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Objective To examine the small antral follicle (SAF) cohort in ovaries of adult rhesus monkeys following consumption of a western-style diet (WSD), with or without chronically elevated androgen levels since before puberty. Design Cholesterol or testosterone (T; n=6/group) implants were placed subcutaneously in female rhesus macaques beginning at 1 yr of age (pre-pubertal), with addition of a WSD (high fat/fructose) at 5.5 yrs (menarche ~2.6 yrs). Ovaries were collected at 7 yrs of age. One ovary/female was embedded in paraffin for morphological and immunohistochemical analyses. The SAFs (<2.5mm) were dissected from the other ovary obtained at/near menses in a subgroup of females (n=3/group), and processed for microarray analyses of the SAF transcriptome. Ovaries of adult monkeys consuming a standard macaque diet (low in fats and sugars) were obtained at similar stages of the menstrual cycle and used as controls for all analyses. Setting National primate research center Animals Adult, female rhesus monkeys (Macaca mulatta) Interventions None Main outcome measures Histological analyses, SAF counts and morphology, protein localization and abundance in SAFs, transcriptome in SAFs (mRNAs) Results Compared to controls, consumption of a WSD, with and without T treatment, increased the numbers of SAFs per ovary, due to the presence of more atretic follicles. Numbers of granulosa cells expressing cellular proliferation markers (pRb and pH3) was greater in healthy SAFs, while numbers of cells expressing the cell cycle inhibitor (p21) was higher in atretic SAFs. Intense CYP17A1 staining was observed in the theca cells of SAFs from WSD+/− T groups, compared to controls. Microarray analyses of the transcriptome in SAFs isolated from WSD and WSD+T treated females and controls consuming a standard diet, identified 1944 genes whose mRNA levels changed ≥2-fold among the three groups. Further analyses identified several gene pathways altered by WSD and/or WSD+T associated with steroid, carbohydrate and lipid metabolism, plus ovarian processes. Alterations in levels of several SAF mRNAs are similar to those observed in follicular cells from women with polycystic ovary syndrome (PCOS). Conclusion These data indicate that consumption of a WSD high in fats and sugars in the presence and absence of chronically elevated T alters the structure and function of SAFs within primate ovaries.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Western-style diet, with and without chronic androgen treatment, alters the number, structure, and function of small antral follicles in ovaries of young adult monkeys

Bishop

et al. 2016

Fertility and Sterility

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“…A cohort of adult, female rhesus macaques (n=25) with a history of normal menstrual cycles were maintained for this study. Hormonal status of each female was monitored for detection of ovulation as previously described [27]; the first day that the serum estradiol (E2) level declines at midcycle below 100 pg/ml with a coincident rise in serum progesterone (P4) was denoted as the first day of the luteal phase, in accordance with previous studies demonstrating the LH-surge occurs the day before E2 levels decline [28]. Monkeys were under the direct care of the ONPRC Department of Comparative Medicine (DCM); all protocols requiring sterile aseptic surgical procedures were performed by trained nonhuman primate (NHP) surgical veterinarians and technicians in the DCM Surgical Services Unit (SSU).…”

Section: Methodsmentioning

confidence: 99%

Dynamics of Immune Cell Types Within the Macaque Corpus Luteum During the Menstrual Cycle: Role of Progesterone1

Bishop

Molskness

et al. 2015

Biology of Reproduction

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The goal of the current study was to characterize the immune cell types within the primate corpus luteum (CL). Luteal tissue was collected from rhesus females at discrete intervals during the luteal phase of the natural menstrual cycle. Dispersed cells were incubated with fluorescently labeled antibodies specific for the immune cell surface proteins CD11b (neutrophils and monocytes/macrophages), CD14 (monocytes/macrophages), CD16 (natural killer [NK] cells), CD20 (B-lymphocytes), and CD3epsilon (T-lymphocytes) for analysis by flow cytometry. Numbers of CD11b-positive (CD11b(+)) and CD14(+) cells increased significantly 3 to 4 days after serum progesterone (P4) concentrations declined below 0.3 ng/ml. CD16(+) cells were the most abundant immune cell type in CL during the mid and mid-late luteal phases and were 3-fold increased 3 to 4 days after serum P4 decreased to baseline levels. CD3epsilon(+) cells tended to increase 3 to 4 days after P4 decline. To determine whether immune cells were upregulated by the loss of luteotropic (LH) support or through loss of LH-dependent steroid milieu, monkeys were assigned to 4 groups: control (no treatment), the GnRH antagonist Antide, Antide plus synthetic progestin (R5020), or Antide plus the estrogen receptor agonists diarylpropionitrile (DPN)/propyl-pyrazole-triol (PPT) during the mid-late luteal phase. Antide treatment increased the numbers of CD11b(+) and CD14(+) cells, whereas progestin, but not estrogen, replacement suppressed the numbers of CD11b(+), CD14(+), and CD16(+) cells. Neither Antide nor steroid replacement altered numbers of CD3epsilon(+) cells. These data suggest that increased numbers of innate immune cells in primate CL after P4 synthesis declines play a role in onset of structural regression of primate CL.

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“…This method prolonged luteal function similar to early pregnancy in rhesus females [23, 24]. Evaluation of ovarian/uterine vascular function (CEUS) and ovarian permeability to albumin (DCE-MRI) was performed the day before and at/near the time of peak P4 response to SEP.…”

Section: Methodsmentioning

confidence: 99%

Intravenous neutralization of vascular endothelial growth factor reduces vascular function/permeability of the ovary and prevents development of OHSS-like symptoms in rhesus monkeys

et al. 2017

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BackgroundOvarian hyperstimulation syndrome (OHSS) is a disorder associated with elevated serum VEGFA following chorionic gonadotropin (hCG) exposure in controlled ovarian stimulation (COS) cycles in women. In this study, we tested the effect of intravenous VEGFA neutralization on OHSS-like symptoms and vascular function in rhesus macaques during COS cycles.MethodsMonkeys (n = 8) were treated with 3 COS protocols and assigned randomly to groups as follows: 1) COS alone (Control, n = 5); 2) COS + VEGF mAb Avastin 19 ± 5 h before hCG (Avastin pre-hCG; n = 6); 3) COS + Avastin 3–4 days post-hCG (Avastin post-hCG; n = 4); 4) COS + Simulated Early Pregnancy (SEP n = 3); or 5) COS + SEP + Avastin (SEP + Avastin n = 3). Follicles were aspirated 36 h post-hCG, fluid was collected from one follicle for analysis of steroid and vascular hormone content. Remaining follicles were aspirated, and luteinized granulosa cells (LGCs) cultured for 24 h. Ovarian/uterine vascular flow (VF) and blood volume (BV) were analyzed by contrast enhanced ultrasound (CEUS) before hCG bolus and 6–8 days post-hCG bolus/time of peak SEP response. Ovarian permeability to albumin was analyzed by Dynamic Contrast Enhanced-MRI (DCE-MRI) post-hCG.ResultsAbdominal fluid was present in 4/5 Control, 2/6 Avastin pre-hCG, and 3/4 Avastin post-hCG females. Neutralization of VEGFA before hCG reduced ovarian VF, BV, and permeability to albumin (P < 0.05), while only ovarian VF and permeability were reduced in Avastin-post hCG group (P < 0.05). There was no effect of Avastin on ovarian vascular function during COS + SEP. VEGF levels in follicular fluid were reduced 78-fold by Avastin pre-hCG, and LGCs exposed to Avastin in vivo also released 4-fold less VEGF into culture media (P < 0.05). Culture medium of LGCs exposed to VEGFA neutralization in vivo had lower levels of P4 and ANGPT1, and an increased ratio of ANGPT2/1 (P < 0.05). Uterine VF was reduced by SEP + Avastin in the basalis/junctional zone (P < 0.05).ConclusionsAvastin treatment before hCG prevents the development of symptoms associated with ovarian hyperstimulation syndrome. In vitro data suggest neutralization of VEGFA alters expression of other vascular factors typically induced by hCG in the luteinizing follicle. Neutralization of VEGFA action alters the vascular function of the basalis zone of the uterus during simulated early pregnancy, indicating a potential effect on embryo implantation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13048-017-0340-5) contains supplementary material, which is available to authorized users.

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Microarray analysis of the primate luteal transcriptome during chorionic gonadotrophin administration simulating early pregnancy

Cited by 18 publications

References 33 publications

Western-style diet, with and without chronic androgen treatment, alters the number, structure, and function of small antral follicles in ovaries of young adult monkeys

Western-style diet, with and without chronic androgen treatment, alters the number, structure, and function of small antral follicles in ovaries of young adult monkeys

Dynamics of Immune Cell Types Within the Macaque Corpus Luteum During the Menstrual Cycle: Role of Progesterone1

Intravenous neutralization of vascular endothelial growth factor reduces vascular function/permeability of the ovary and prevents development of OHSS-like symptoms in rhesus monkeys

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