Differential regulation of cumulus cell transcription during oocyte maturation in vivo and in vitro

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HAS2-AS1 is a novel LH/hCG target gene regulating HAS2 expression and enhancing cumulus cells migration

Abstract Background The cumulus expansion process is one of the LH mediated ovulatory processes. Hyaluronan synthase 2 (HAS2) regulates the synthesis of hyaluronic acid, the main component of the cumulus expansion process. Recently, the lncRNA HAS2 antisense RNA 1 (HAS2-AS1) was identified in our global transcriptome RNA-sequencing of novel ovulation associated genes. The role of HAS2-AS1 in HAS2 regulation w.as studied previously with contradictive results in different models but not in the ovary. Taken together the induction of HAS2-AS1 and the important role of HAS2 in the cumulus expansion process, we hypothesize that HAS2-AS1 regulate HAS2 expression and function in the ovary. Therefore we undertook to study the expression, regulation, and possible functional role of HAS2-AS1 in the human ovary. Results HAS2-AS1, located within the HAS2 gene that was highly regulated in our library. We found that HAS2-AS1 express mainly in cumulus cells (CCs). Furthermore, HAS2-AS1 showed low expression in immature CCs and a significant increase expression in mature CCs. Functional studies reveal that inhibition of HAS2-AS1 by siRNA caused decrease expression of HAS2. Furthermore, inhibition of HAS2-AS1 by siRNA results in decrease migration of granulosa cells. Conclusions Our results suggest that HAS2-AS1 is an LH/hCG target gene that plays a positive role in HAS2 expression and thus might play a role in regulating cumulus expansion and migration

GnRH Agonist vs. hCG for Triggering of Ovulation – Differential Effects on Gene Expression in Human Granulosa Cells

<div><p>Objective</p><p>To investigate the mRNA expression of genes related to steroidogenesis and OHSS in granulosa cells (GCs) of patients triggered with GnRH agonist compared to patients triggered with hCG.</p><p>Design</p><p>Mural GCs were obtained at the time of oocyte retrieval and gene expression was analyzed using quantitative real time RT-PCR.</p><p>Settings</p><p>Single center, case control study.</p><p>Patient(s)</p><p>24 women who were treated with GnRH agonist or hCG for triggering of ovulation.</p><p>Interventions</p><p>GC collection.</p><p>Main Outcome Measure(s)</p><p>The expression of genes related to steroidogenesis and OHSS in mural GCs</p><p>Results</p><p>The fertilization rate was similar in the two groups. The mRNA expression of CYP19A1 (0.50 vs 1, arbitrary unit), CYP11A1 (0.6 vs. 1) and 3 beta hydroxysteroid-dehydrogenase (0.39 vs 1) was significantly lower in the GnRH group. The expression of VEGF (0.74 vs. 1) and inhibin β B (0.38 vs 1) was lower in the GnRH analog triggered group.</p><p>Conclusion</p><p>Expression of genes related to steroidogenesis is lower at the time of oocyte retrieval in patients triggered with GnRH agonist. The decreased expression of VEGF and inhibin β B in the GnRH agonist group can explain the mechanism of early OHSS prevention.</p></div

Expression of STAR in mGCs obtained from IVF cycles separated according to ovulation triggering.

<p>Expression of STAR in mGCs obtained from IVF cycles separated according to ovulation triggering.</p

Characteristics of the patients and IVF procedures included in this study.

<p>Characteristics of the patients and IVF procedures included in this study.</p

Expression of epiregulin in mGCs obtained from IVF cycles separated according to ovulation triggering.

<p>Expression of epiregulin in mGCs obtained from IVF cycles separated according to ovulation triggering.</p

Expression of VEGF in mGCs obtained from IVF cycles separated according to ovulation triggering.

<p>Expression of VEGF in mGCs obtained from IVF cycles separated according to ovulation triggering.</p