Study On The Gonad-related Hormones And The Genes Of Penaeus Monodon

Penaeus monodon,commonly known as grass shrimp and tiger shrimp,is an important economic species for marine aquaculture in China.At present,the whole artificial reproducing technology of Penaeus monodon has made tremendous breakthroughs and progress.But in the reproducing process,the traditional facilitating maturation method of cutting the eyestalks often causes certain deaths.Therefore,it is very important to find new methods for inducing gonad maturation and study the regulation hormones related to gonad development.In addition to hormone regulation,the regulation of gonad development in Penaeus monodon is also influenced and regulated by a variety of neurotransmitters.However,there are few reports on mammalian sex hormones which are directly act on gonad organs.Gonadotropin-releasing hormone is an important neurohormone that plays an important role in animal reproducing.This study by Liquid Chromatography technique of Penaeus monodon in the ovarian tissue and brain tissue extract separation of Gn RH,identifying ovarian tissue and brain Gn RH in crude extracts of existence,and to explore the separation method.A hormone similar to Follicle-Stimulating Homne(FSH)found in mammals,named Crustean female sex hormone(CFSH),is thought to be related to the regulation of crustacean ovarian development.Therefore,we cloned the CFSH gene of Penaeus monodon in this experiment and explored its stress response mechanism.The results are as follows: 1 Gonadotropin-releasing hormone preliminary separation The presence of Gn RH was identified from the crude extracts of ovarian tissue and brain tissue of Penaeus monodon by dot immunoassay.After a preliminary attempt to separate the crude extract,it was found that the gradient of mobile phase B from 5% to 35% can obtain a better separation chromatogram.The identification of Gn RH analogues in crude ovarian extract provides a new reference basis for the separation and purification of Penaeus monodon Gn RH.2 Molecular cloning and multifunction exploration of CFSH gene in Penaeus monodonThe ORF and 3'UTR of Pm CFSH [Penaeus monodon Crustacean female sex hormone)were cloned by RACE(Rapid-amplification of c DNA ends)method.A protein of 214 aa was predicted to encode,which contained a signal peptide and an interleukin 17E(IL-17E)domain relating to immune response.The q RT-PCR results show that the Pm CFSH was expressed in various tissues,among which the highest expression level was found in the abdominal nerve tissue;the expression level gradually increased from fertilized egg stage to larval stage,and the expression level reached the highest at the larval stage;During the ovarian development Stage ? to ?,it gradually increased as the ovary matured;In view of the IL-17 domains of Pm CFSH,we did bacteria stimulate experiments to explore its response in immune response mechanisms.Harvey vibrio and expression of staphylococcus aureus bacteria are able to raise Pm CFSH.The study reveals that Pm CFSH not only participates in the regulation of larval development and gonad maturity,but also has an immune response to bacterial stress,reflecting the "multifunctional" phenomenon of hormones.3 Interference experiment of crustacean female hormone CFSH geneThe interference results showed that the decreased expression of Pm CFSH may promote the development of male gonadal gonad tissue and spermatogenesis.However,from the observation of tissues,interference with the Pm CFSH gene will lead to an increase in the rate of sperm malformation in male sperm pods,but the effect on the development of the gonads of female Penaeus monodon needs further investigation.