Studies On Cranial Nerve Regulation Genes Mining And Its Expressed Molecular Mechanism In Ejaculation Process Of Male Rats

Ejaculation is the ejection of semen from the male reproductory tract,and the basic phenomenon of life which matains sex harmony and life forms to the continuation in human or other mammals.As to humans,ejaculation is considered to be the most amazing biological phenomenon of life.Ejaculatory dysfunction including prosermia and retardation affect the fertility and quality of life.Molecular mechanism of regulation in animal brain during ejaculation not only opens up new way to understand the physical laws of animal ejaculation,but also provides scientific evidence for clinical research.In this paper,ejaculation-re la ted genes in brain will be excavated in the process of ejaculation of male rats by sequencing,aiming to provide a scientific basis for exploring the molecular mechanism in rat brain regulation during ejaculation.1.A simple and convenient experimental mating model in male rat was established,which could be suitable for the subject-related research.2.The effects of different dose of 8-OH-DPAT and dapoxetine at different dose on the ejaculation in male rat were evaluated.The results showed that 8-OH-DPAT(0.5mg/kg)could facilitate the ejaculation,while dapoxetine(60mg/kg)delayed the ejaculation in male rats.Therefor,two does of them were chose to the followed transcriptom experiment.3.In Solexa high-throughput sequencing technology-based transcriptome studying(Six transriptome groups:control-CK,The fourth intromission一CR4,Ejaculation一EJ,8-OH-DPAT to ejaculation-DP AT,dapoxetine to ejaculation-DAP and postejaculation interval 1 min-PEI1),by mutual comparison between transcriptome and statistical differences in genes,the major result showed:the expression pattern of 42genes were up-regulated and 97 genes were down-regulated in CR4/CK;the expression pattern of 66 genes were up-regulated and 191 genes were down-regulated in EJ/CK;the expression pattern of 91 genes were up-regulated and 258 genes were down-regulated in DPAT/CK;the expression pattern of 52 genes were up-regulated and 155 genes were down-regulated in DAP/CK;the expression pattern of 39 genes were up-regulated and 91 genes were down-regulated in PEI1/CK;the expression pattern of 112 genes were up-regulated and 191 genes were down-regulated in EJ/CR4;the expression pattern of 128 genes were up-regulated and 194 genes were down-regulated in PEI1/EJ;the expression pattern of 264 genes were up-regulated and 487 genes were down-regulated in DPAT/EJ;the expression pattern of 162 genes were up-regulated and 211 genes were down-regulated in DAP/EJ.4.The differences in gene expression level and its change law were analyzed,we speculate that the Dnd4,Tph1,Chrna3,Chrnb4,Oxt,Prl,Avp,Pomc,Gabra6,Gabrrl and Cacnalf,a total of 11 genes may be involved in regulating the biological behavior of ejaculation.In addition,Drd1 and Slc6a3 may involve the molecular mechanism of 8-OH-DPAT promoting ejaculation,and Drd4 involved the molecular mechanism of dopaxetine delaying ejaculation.5.The 11 genes were detected by using qRT-PCR technique in this validate,result showed that:changes in the relative expression levels of 11 genes were similar to the transcriptome.This showed transcriptome data in our experiment were reliable and accurate.6.Based on the comprehensive analysis of the transient expressive ejaculation-related genes in male rats,we hypothesis:When external stimulated signals acting on the central nervous,stimulated related genes including neurotransmitter receptors,ion channels,the Philippian peptide precursor,peptide hormones,etc.,were involved in the regulation of ejaculation.They directly or indirectly,positive or negative feedback to the central nervous system by translating functional proteins or enzymes,and formed the brain control loop to complete regulating in the process of ejaculation.7.We analyzed gene homology of 11 ejaculation related genes through bioinformatics,found Tph1,Chrna3,Oxt,Gabra6,Gabrr1 and Cacnalf more conservative,while Drd4,Chrnb4,Pr1,Avp and Pomc less conservative.By the average homology analysis,they may evolve faster in humans.In this paper,a transcriptome approach using high-throughput Solexa sequencing technology was applied to explore the molecular mechanism in rat brain regulation during ejaculation.A lot of genetic information were obtained from male rats,and filtered out the possible genes involved in regulatin ejaculation It laied a solid foundation for the study of the molecular mechanisms of ejaculation.