Inhibitory effect of retinoic acid on the development of immature porcine granulosa cells to mature cells

Abstract

The present study investigated the effect of retinoic acid (RA) on the differentiation of granulosa cells prepared from porcine ovaries. The granulosa cells were precultured for 15 h, then cultured for 48 h with FSH and further treated for 24 h with LH in order to induce their transformation into luteal cells. After the cells had been exposed to 1 µM retinoids (RA, retinal and retinol) for 87 h, analysis of the LH receptor mRNA expression, an indicator of granulosa cell differentiation, was carried out by using semiquantitative RT-PCR. The results showed that there was a decrease in LH receptor mRNA levels, and that RA had a more potent effect on these levels than the other two retinoids. When cells were exposed to RA in the immature stage (before the addition of FSH) or the early stage of development (0-24 h after the addition of FSH), expression of LH receptor mRNA was greatly diminished. When the immature cells were cultured for 15 h with RA, then washed and cultured for 48 h with FSH and for 24 h with LH, the expression of LH receptor mRNA was not reversed. In the differentiated cells (24 h after the addition of FSH), however, RA no longer had any inhibitory effect. When the immature cells were exposed to RA, FSH-induced expression of c-fos mRNA was markedly decreased. In contrast, expression of c-jun and activating transcription factor-4 mRNAs remained constant. However, the expression of c-fos mRNA was not decreased by forskolin. The results indicate that RA is a potent inhibitor in the immature stage of porcine granulosa cell differentiation, probably through decreased expression of FSH receptor, but that RA does not inhibit differentiation in the mature stage of the cells.

9‐cis‐retinoic acid during in vitro maturation improves development of the bovine oocyte and increases midkine but not IGF‐I expression in cumulus‐granulosa cells

“…The MK gene product could be an intermediary in the above reported developmental phenomena (Griffith and Zile, 2000). In rat granulosa cells, 0.3 mM RA resulted in a two fold increase in MK mRNA levels (Minegishi et al, 1996), and 1 mM RA altered LH receptor mRNA expression (Hattori et al, 2000). The embryotrophic effects of MK reported by Ikeda et al (2000a,b) reproduce in part the effect of RA within the present work.…”

Supplementation of 9-cis retinoic acid in the in vitro maturation medium increase blastocyst development rate and quality

“…These results are consistent with previous reports (Deb et al, 2011;Gómez et al, 2003). The beneficial effects of RA during IVM are mediated by enhancing oocyte maturation through its effect on FSH or LH receptor expression (Hattori et al, 2000), increasing mRNA quality and processing (Gomez et al, 2004), growth factors signaling by endogenous oxidative-stress protection mechanism (Guerin, 2001) and /or prevention of apoptosis (Deb et al, 2011). …”

Retinoid receptor-specific agonists regulate bovine in vitro early embryonic development, differentiation and expression of genes related to cell cycle arrest and apoptosis

“…These authors found antiproliferative and pro-apoptotic effects that mainly targeted the ICM [17]. Concentrations of ATRA between 0.3 mM and 1 mM are commonly encountered in the literature on reproductive cells [9,18,19]. Finally, a concentration of 1 mM ATRA during IVM can improve blastocyst development (Ikeda S, personal communication).…”