Effect Of RFRP-3 On Reproductive Function Of Rat Epididymis

GnIH is a novel neuropeptide with an RF amide structure at the C-terminus found in the brain of a Japanese research team.Later,neuropeptides with an RF amide structure were also found on mammals.This RF amide peptide was named RF-Related Peptide(RFRP),and RFRP-3 was consistent with the physiological role of GnIH in birds.At present,there are few studies on the effect of RFRP-3 on the reproductive function of male animals at epididymal level.In this study,we examined the expression of RFRP-3 and GPR147 mRNAs in epididymis of rats at different stages of development.Then we examined the effects of different doses of RFRP-3 on the morphology,apoptosis,and autophagy-related protein expression in epididymis of rats.Investigate the effect of RFRP-3 on the reproductive function of epididymis in rats.This study can be divided into three parts:(1)Detect the changes of expression of RFRP-3 and GPR147 mRNAs in the epididymis of 20,40,60,and 80 days old rats(corresponding to P20,P40,P60,and P80,respectively).The results showed that the expression levels of RFRP-3 and GPR147 mRNAs fluctuate with the development of epididymis in rats,and the expression levels of RFRP-3 and GPR147 mRNAs were highest at P60,suggesting that RFRP-3 and its receptor may be involved in rat epididymis.Develops and regulates its reproductive activity.(2)Through in vivo experiments,observe the effect of RFRP-3 on rat epididymis histology.Rats were divided into control and 0.1μg,1μg and 10μg/day different dose groups,6 per group.The rats were injected intraperitoneally with RFRP-3 for 7 consecutive days,and one side of the epididymis was removed and paraffin sections were made and HE staining was performed under a microscope.The results showed that RFRP-3 can cause epididymal epithelial cell abnormalities in a dose-dependent manner and significantly reduce the number of spermatozoa in the epididymis,suggesting that RFRP-3 induces functional decline in the epididymis.(3)The mechanism of RFPR-3 induced functional decline of epididymis in rats was investigated by examining the levels of apoptosis and autophagy in the epididymis of rats.On the one hand,we have detected that RFRP-3 can induce increased activity of Caspase-3 in epididymal tissue.On the other hand,Western blot analysis revealed that different doses of RFRP-3 caused conflicting epididymal apoptosis and expression of autophagy marker proteins,suggesting a complex interaction between apoptosis and autophagy.In summary,RFRP-3 plays an important regulatory role in the reproductive function of male epididymis,probably because RFRP-3 mediates epididymal apoptosis and autophagy.