Effects of kisspeptin-10 on progesterone secretion in cultured chicken ovarian granulosa cells from preovulatory (F1-F3) follicles

The effect of kisspeptin-10 (Kp-10) on the secretion of progesterone (P-4) was investigated in cultured granulosa cells from F-1 to F-3 follicles of hens. The results showed that granulosa cells were stained with clear signals for kisspeptin using immunocytochemistry with the specific antibody against Kp-10. Among 10, 100 and 1000 nM concentrations tested, 100 nM Kp-10 treated for 24 h significantly increased P-4 secretion in granulosa cells from F-1 to F-3 follicles. After 24 h and 48 h of treatment, 100 nM Kp-10 showed a significant increase in P-4 secretion, while after 72 h of treatment 134 secretion was markedly decreased by Kp-10 compared to the control group (P < 0.05). F-1 and F-2/3 cells treated with 100 nM Kp-10 for 24 h showed significantly increased viability (P < 0.05) and which was in parallel to a marked increase in 134 secretion (P < 0.01). Real-time PCR results showed that the gene expression of the steroidogenic acute regulatory protein (StAR), cytochrome P450 side-chain cleavage (P450scc) and the enzyme 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) in F-1 and F-2/3 granulosa cells was significantly up-regulated by 24 h-100 nM Kp-10 treatment (P < 0.05 versus P < 0.01, respectively). However, there was no significant difference in StAR, P450scc and 3 beta-HSD protein content between control and the Kp-10 treated group (P > 0.05). These results indicate that Kp-10 stimulates 134 secretion in cultured chicken granulosa cells, which was associated with an up-regulation in StAR, P450scc and 3 beta-HSD gene transcription. (C) 2011 Elsevier Inc. All rights reserved.