Introduction Previous studies suggest that Progesterone Receptor Membrane Component 1 (PGRMC1) plays an essential role during bovine oocyte meiosis, since it 1) localizes to the centromeres at metaphase-I and II and 2) concentrates between the separating chromosomes at ana/telophase-I. Moreover, injection of an antibody to PGRMC1 significantly impairs completion of meiosis. The aim of the present study is to expand these findings by using small-interfering RNA (RNAi) mediated gene silencing. Methods Cumulus-Oocytes Complexs were microinjected to deliver PGRMC1 or CTRL-RNAi into the cytoplasm of oocytes, kept in meiotic arrest for 18h with 10µM cilostamide and then in vitro-matured for 24h (IVM). After IVM, efficacy in depleting PGRMC1 expression was assessed by quantitative RT-PCR. Finally, the oocyte capability to extrude the first polar body (PBI) and the morphology of the MII plates were assessed. Results and Discussion PGRMC1 expression following PGRMC1-RNAi treatment was significantly reduced by 30%. This was accompanied by a 22% reduction of the oocytes that extruded the PBI (p<0.05). Surprisingly, PGRMC1-RNAi treatment did not affect MII plate formation or morphology. However, a significantly higher proportion of PGRMC1-RNAi injected oocytes possessed clumps of DNA scattered throughout the ooplasm in addition to the MII plate (p<0.05). This is consistent with PGRMC1 localization at the midbody and with a putative role in cytokinesis. We hypothesize that lower PGRMC1 expression impairs the process of PBI formation. As a consequence, DNA that should be extruded with the PBI is retained in the cytoplasm and degraded. Funding: FP7-PEOPLE-2011-CIG, contract n.:303640-Pro-Ovum.
Insight into Progesterone Receptor Membrane Component 1 (PGRMC1) action during bovine oocyte meiosis by means of small-interfering RNA mediated gene silencing / V. Lodde, I. Tessaro, F. Raucci, F. Franciosi, F. Marchese, S. Modina, J.J. Peluso, A.M. Luciano. - In: REPRODUCTION ABSTRACTS. - ISSN 2052-1472. - 2014:1(2014), pp. P044.1-P044.1. ((Intervento presentato al convegno World Congress of Reproductive Biology tenutosi a Edinburg nel 2014.
|Titolo:||Insight into Progesterone Receptor Membrane Component 1 (PGRMC1) action during bovine oocyte meiosis by means of small-interfering RNA mediated gene silencing|
LODDE, VALENTINA (Primo) (Corresponding)
TESSARO, IRENE (Secondo)
LUCIANO, ALBERTO MARIA (Ultimo)
|Settore Scientifico Disciplinare:||Settore VET/01 - Anatomia degli Animali Domestici|
Settore VET/02 - Fisiologia Veterinaria
Settore BIO/06 - Anatomia Comparata e Citologia
|Progetto:||Role of Progesterone Receptor Membrane Component-1 in oogenesis and mammalian fertility|
|Data di pubblicazione:||2014|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1530/repabs.1.P044|
|Appare nelle tipologie:||01 - Articolo su periodico|