Óscar Peña

Angiotensin II induces apoptosis in human mural granulosa-lutein cells, but not in cumulus cells.

OBJECTIVE To test whether angiotensin II (AngII) could modulate apoptosis of human granulosa-lutein (GL) cells from gonadotropin-stimulated follicles. DESIGN In vitro assays on mural and cumulus granulosa cells. SETTING University laboratory and private IVF practice. PATIENT(S) One hundred six consecutive women undergoing 113 IVF cycles. INTERVENTION(S) Purified human GL mural or cumulus cells were cultured in serum-free media in the presence or absence of AngII with or without the AngII receptor blockers saralasin and CGP42112A. MAIN OUTCOME MEASURE(S) Detection of apoptosis using a fluorescent in situ marker for activated caspases. RESULT(S) Mural cells had approximately eightfold the amount of apoptosis compared with cumulus cells (average 0.23 vs. <0.03, respectively). With mural cells, AngII increased GL cell apoptosis versus untreated control samples (AngII 10(-)11 mol/L +6.5%; AngII 10(-9) mol/L +13.3%, and AngII 10(-7) mol/L +11.3%), an effect which was blocked by concurrent incubation with AngII receptor blockers. The AngII receptor blockers produced a significant decrease of apoptosis compared with control cultures (saralasin: 19.4%; CGP42112A: 28.9%). Neither AngII nor blockers had effect on cumulus cells. CONCLUSION(S) Preovulatory concentrations of AngII, most likely via AT2 receptors, increase apoptosis of cultured mural GL cells but have no effect on cumulus cells. Granulosa cells appear to be differentially regulated by AngII.

FSH receptor, KL1/2, P450, and PAPP genes in granulosa-lutein cells from in vitro fertilization patients show a different expression pattern depending on the infertility diagnosis

OBJECTIVE To study the expression of mRNA for Kit ligand 1 (KL1), KL2, FSH receptor (FSHR), pregnancy-associated plasma protein (PAPP) and P450 in granulosa-lutein cells from IVF patients and its relationship with the infertility diagnosis and IVF outcome. DESIGN In vitro assays. SETTING University laboratory and private IVF center. PATIENT(S) A total of 113 women undergoing IVF. INTERVENTION(S) Quantitative reverse-transcription polymerase chain reaction in granulosa-lutein cells from pooled follicles. MAIN OUTCOME MEASURE(S) Expression of mRNA for KL1, KL2, FSHR, PAPP and P450 in infertility patients with different infertility diagnoses. RESULT(S) Infertile patients with healthy ovaries show positive correlations among KL1, KL2, FSHR, PAPP, and P450 gene expression levels. In patients with ovarian disease, the correlation between KL1 and KL2 gene expression is maintained, but correlations between KL1/KL2 and FSHR, PAPP and P450 are not present except for KL1/FSHR in endometriosis and KL2/P450 in polycystic ovary syndrome. FSHR/KL1 and FSHR/KL2 expression correlates positively only in women who became pregnant. CONCLUSION(S) Findings in healthy human ovaries agree with the feedback model of bone morphogenetic protein 15-KL1/KL2 cross-talk between oocyte and granulosa cells described in other species. The complex relationship between the expression of these genes seems to be disrupted in patients with infertility of ovarian origin. A normal pattern of gene expression and feedback seems to be associated with pregnancy.