Marli Amin1, Ariel Simerman, Michele Cho, Prapti Singh, Christine Briton-Jones, David Hill, Tristan Grogan, David Elashoff, Nigel J Clarke, Gregorio D Chazenbalk, Daniel A Dumesic. 1. Department of Obstetrics and Gynecology (M.A., A.S., M.C., P.S., G.D.C., D.A.D.), David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, California 90024; Department of Medicine Statistics Core (T.G., D.E.), David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, Los Angeles, California 90095; Quest Diagnostics Nichols Institute (N.J.C.), San Juan Capistrano, California 92675; and ART Reproductive Center (C.B.-J., D.H.), Beverly Hills, California 90210.
Abstract
CONTEXT: Mineralocorticoid synthesis by the nonhuman primate periovulatory follicle enhances luteinization. Whether a similar event occurs in women undergoing in vitro fertilization (IVF) is unknown. OBJECTIVE: The objective of the study was to determine whether human luteinized granulosa cells (LGCs) produce mineralocorticoids derived from 21-hydroxylase activity and also express mRNA for 21-hydroxylase and the mineralocorticoid receptor. DESIGN: This was a prospective cohort study. SETTING: The study was conducted at an academic center. PATIENTS: LGC lipid content and follicle fluid (FF) hormone analysis was performed on 27 nonobese IVF women. LGCs from six additional nonobese IVF women were used for gene expression studies. INTERVENTION: At oocyte retrieval, FF was aspirated from the first follicle (≥16 mm in size) of each ovary and pooled LGCs were collected. MAIN OUTCOME MEASURES: FF steroid analysis was performed by liquid chromatography-tandem mass spectrometry. LGCs were stained with lipid fluorescent dye BODIPY FL C16 to estimate lipid content by confocal microscopy as a cholesterol source for steroidogenesis in vivo. Quantitative real-time PCR was performed using LGCs to detect 21-hydroxylase and mineralocorticoid receptor mRNA expression. Pearson correlation coefficients determined associations between FF steroid levels and LGC lipid content. RESULTS: FF levels of the 21-hydroxylase-derived steroids, 11-deoxycorticosterone [DOC, 39.97, median (13.94-63.02) ng/mL] and 11-deoxycortisol [11DOC, 2.07 (0.69-5.01) ng/mL], along with the 21-hydroxylase precursor 17-hydroxyprogesterone [1268.21 (493.26-3558.39) ng/mL], positively correlated with LGC lipid content (84 ± 43 fluorescent units/sample) (P ≤ .05, all steroids). 21-Hydroxylase and mineralocorticoid receptor mRNA expression was detected in LGCs. CONCLUSIONS: Human LGCs likely synthesize 21-hydroxylase-derived mineralocorticoids from cholesterol-containing lipid in vivo to promote postovulatory luteinization via mineralocorticoid receptor-mediated events.
CONTEXT: Mineralocorticoid synthesis by the nonhuman primate periovulatory follicle enhances luteinization. Whether a similar event occurs in women undergoing in vitro fertilization (IVF) is unknown. OBJECTIVE: The objective of the study was to determine whether human luteinized granulosa cells (LGCs) produce mineralocorticoids derived from 21-hydroxylase activity and also express mRNA for 21-hydroxylase and the mineralocorticoid receptor. DESIGN: This was a prospective cohort study. SETTING: The study was conducted at an academic center. PATIENTS: LGC lipid content and follicle fluid (FF) hormone analysis was performed on 27 nonobese IVFwomen. LGCs from six additional nonobese IVFwomen were used for gene expression studies. INTERVENTION: At oocyte retrieval, FF was aspirated from the first follicle (≥16 mm in size) of each ovary and pooled LGCs were collected. MAIN OUTCOME MEASURES: FF steroid analysis was performed by liquid chromatography-tandem mass spectrometry. LGCs were stained with lipid fluorescent dye BODIPY FL C16 to estimate lipid content by confocal microscopy as a cholesterol source for steroidogenesis in vivo. Quantitative real-time PCR was performed using LGCs to detect 21-hydroxylase and mineralocorticoid receptor mRNA expression. Pearson correlation coefficients determined associations between FF steroid levels and LGC lipid content. RESULTS: FF levels of the 21-hydroxylase-derived steroids, 11-deoxycorticosterone [DOC, 39.97, median (13.94-63.02) ng/mL] and 11-deoxycortisol [11DOC, 2.07 (0.69-5.01) ng/mL], along with the 21-hydroxylase precursor 17-hydroxyprogesterone [1268.21 (493.26-3558.39) ng/mL], positively correlated with LGC lipid content (84 ± 43 fluorescent units/sample) (P ≤ .05, all steroids). 21-Hydroxylase and mineralocorticoid receptor mRNA expression was detected in LGCs. CONCLUSIONS:Human LGCs likely synthesize 21-hydroxylase-derived mineralocorticoids from cholesterol-containing lipid in vivo to promote postovulatory luteinization via mineralocorticoid receptor-mediated events.
Authors: A E Michael; M Evagelatou; D P Norgate; R J Clarke; J W Antoniw; B A Stedman; A Brennan; R Welsby; I Bujalska; P M Stewart; B A Cooke Journal: Mol Cell Endocrinol Date: 1997-09-19 Impact factor: 4.102
Authors: Ariel A Simerman; David L Hill; Tristan R Grogan; David Elashoff; Nigel J Clarke; Ellen H Goldstein; Alexa N Manrriquez; Gregorio D Chazenbalk; Daniel A Dumesic Journal: Fertil Steril Date: 2014-10-24 Impact factor: 7.329