| Immune System plays an important role for identifying and eliminating foreign-body antigen. Female reproductive system is the key organ in breeding new life,its local immunity is regulated by the nervous and endocrine.To explore whether the TH1-type cytokines can influence the physiological state of the female reproductive system by both sympathetic and sensory nerve pathway,while these cytokines can play a biological activity after combining with their corresponding receptor. In this study, we used molecular biology and proteomics approach to explore the expression and localization of IL18Rαin inferior mesenteric ganglion(IMG) and lumbar dorsal root ganglions(LDRGs) in female goats, it was proved that IL18 could act on the visceral afferent and sympathetic efferent neurons by direct or indirect means , and then affecting the reproductive physiology of uterine activity, in order to provide the morphological evidence for studying the neural immune regulation of female reproductive system.The main experimental results are as follows:1. Goat IL18Rαgene partial cDNA sequences was initially identified in the IMG and LDRGs by molecular biology methods using bovine,human and porcine IL18Rαas the seed. The partial sequence for goat IL18RαcDNA was 447 bp, and predicted to have 148 amino acids.2. Bioinformatics software was used to blast goat IL18Rαgene sequence and the amino acid sequence homology with other species.Goat IL18Rαdisplayed extensive homology with IL18Rαof other species. The predicted amino acid sequence shared 100%, 90% and 88% homology with bovine, human, and porcine IL18Rα, respectively ,and gene sequence homology comparison showed 91%, 88%, 87% homology with bovine, human, and porcine IL18Rα,respectively.3. The cloning of the IL18Rαpartial cDNA as a template,digoxigenin-labelled RNA probes were synthesized by reverse transcription in virto for in situ hybridization. Expression and localization of IL18RαmRNA was detected in IMG and LDRGs,and the results showed that IL18RαmRNA mainly located in the cytoplasm of neurons, satellite cells and connective tissue between neurons in the IMG and IL18RαmRNA was expressed neuronal nuclear membrane, cytoplasm of neurons, satellite cells, some neurogliocytes and medullated fibers in LDRGs.4. It is verified that IL18Rαprotein existed in the IMG and LDRGs by Werstern Blot and immunohistochemical SP method, and interesting is this result and the expression of its mRNA remained the same basically.In the IMG, positive products of IL18Rαmainly expressed in the cytoplasm of neurons, satellite cells and some positive particles between neurons; in DRG, nuclear membrane,neuronal cytoplasm,satellite cells, some neurogliocytes and medullated fibers showed positive expression, and nucleolus was negative. |
