title: 21-Hydroxylase-derived steroids in follicles of nonobese women undergoing ovarian stimulation for in vitro fertilization (IVF) positively correlate with lipid content of luteinized granulosa cells (LGCs) as a source of cholesterol for steroid synthesis.
abstract: BACKGROUND
  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.
  
  
  METHODS
  This was a prospective cohort study.
  
  
  METHODS
  The study was conducted at an academic center.
  
  
  METHODS
  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.
  
  
  METHODS
  At oocyte retrieval, FF was aspirated from the first follicle (≥16 mm in size) of each ovary and pooled LGCs were collected.
  
  
  METHODS
  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.affiliation: 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.
authors.forename: Marli
authors.initials: M
authors.lastname: Amin
authors.forename: Ariel
authors.initials: A
authors.lastname: Simerman
authors.forename: Michele
authors.initials: M
authors.lastname: Cho
authors.forename: Prapti
authors.initials: P
authors.lastname: Singh
authors.forename: Christine
authors.initials: C
authors.lastname: Briton-Jones
authors.forename: David
authors.initials: D
authors.lastname: Hill
authors.forename: Tristan
authors.initials: T
authors.lastname: Grogan
authors.forename: David
authors.initials: D
authors.lastname: Elashoff
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Clarke
authors.forename: Gregorio D
authors.initials: GD
authors.lastname: Chazenbalk
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Dumesic
mesh_terms: D000328:Adult; D002784:Cholesterol / Q000378:metabolism; D015331:Cohort Studies; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000737:chemistry / Q000378:metabolism; D006107:Granulosa Cells / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D056865:Ideal Body Weight; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis; D034461:Luteinization / Q000502:physiology; D008901:Mineralocorticoids / Q000032:analysis / Q000378:metabolism*; D006080:Ovarian Follicle / Q000737:chemistry / Q000378:metabolism*; D010062:Ovulation Induction; D013255:Steroid 21-Hydroxylase / Q000235:genetics / Q000378:metabolism*
chemical_list: D008055:Lipids; D008901:Mineralocorticoids; D002784:Cholesterol; C531510:CYP21A2 protein, human; D013255:Steroid 21-Hydroxylase