Gonadotropin-releasing hormone messenger ribonucleic acid and protein expression in Vero cells

Purpose: Interleukin-1 (IL-1) is a major regulator of local cellular intera ctions during embryonic implantation. We hypothesized that gonadotropin-rel easing hormone (GnRH) may also play a role in the embryonic/epithelial dial ogue during early implantation. To examine this hypothesis, we examined the ability of IL-1 to regulate GnRH mRNA and protein expression in Vero cells . Methods Viable Vero cells (1 x 10(5)/well) were cultured in multiple-well t issue culture plates for in vitro studies and in 4-well chamber slides for in immunohistochemical study. Confluent Vero cells were cultured with incre asing concentrations of recombinant human IL-1 beta for an additional 24 he Vero cell expression of GnRH and GnRH receptor mRNAs was measured with pol ymerase chain reaction (PCR) and nested PCR, respectively GnRH protein expr ession was validated by immunohistochemistry study. The quantitative lever of GnRH mRNA expression regulated by IL-1 beta in Vero cells was determined by quantitative competitive PCR (QC PCR) with standard curve methodology. Results: RT-PCR revealed beta -actin, GnRH, and GnRH receptor mRNA expressi on in Vero cell cultures. Immunostaining confirmed the presence of GnRH pro tein in Vero cells. Quantitative PCR demonstrated IL-1 beta up-regulation o f Vero cell GnRH mRNA expression (p < 0.05). Conclusions: These results suggest that Vero cell mRNA and protein expressi on of GnRH may play a substantial role in early embryo/epithelial dialogue during embryo coculture, with nn embryotrophic effect due to expression of GnRH by Vero cells.