Zhang Kaizhao, Tao Pan, Liu Jianxin, Wang Qingnan, Ge Shikun, Ning Zhangyong
College of Veterinary Medicine, South China Agricultural University , Guangzhou 510642 , People's Republic of China.
Acta Vet Hung. 2017 Sep;65(3):402-416. doi: 10.1556/004.2017.038.
The NOD-like receptor protein 3 (NLRP3) inflammasome comprised of NLRP3, ASC and caspase-1 plays an important role in the inflammatory and innate immune response. However, little is known about the expression pattern and histological distribution of these genes in goat. Here, we first cloned the fulllength cDNAs of the NLRP3, ASC and caspase-1 genes of Hainan black goat and produced their polyclonal antibodies. Tissue-specific expression and histological distribution of these genes were analysed. Phylogenetic analysis revealed that these three goat genes had high homology with Bos taurus genes and low homology with avian or fish genes. After immunisations with these recombinant Histagged proteins, the titres of antiserum were higher than 1:1024 and purified IgG was obtained. These three genes were expressed in all examined tissues, the mRNA expression level of NLRP3 and caspase-1 was most abundant in the spleen and mesenteric lymph nodes (MLNs), while ASC was primary expressed in the liver, spleen and kidney. The histological distribution of NLRP3, ASC and caspase-1 was detected in myocardial cells, hepatocytes, focal lymphocytes, bronchiolar epithelial cells, renal tubular epithelial cells, cortical neurons and endothelial cells of the germinal centres in the MLNs. These results will be helpful in further investigations into the function of the NLRP3 inflammasome and in elucidating its role in caprine inflammatory diseases.
由NLRP3、ASC和半胱天冬酶-1组成的NOD样受体蛋白3(NLRP3)炎性小体在炎症和固有免疫反应中起重要作用。然而,关于这些基因在山羊中的表达模式和组织学分布却知之甚少。在此,我们首先克隆了海南黑山羊NLRP3、ASC和半胱天冬酶-1基因的全长cDNA,并制备了它们的多克隆抗体。分析了这些基因的组织特异性表达和组织学分布。系统发育分析表明,这三个山羊基因与牛基因具有高度同源性,而与禽类或鱼类基因的同源性较低。用这些重组的带有组氨酸标签的蛋白免疫后,抗血清滴度高于1:1024,并获得了纯化的IgG。这三个基因在所有检测组织中均有表达,NLRP3和半胱天冬酶-1的mRNA表达水平在脾脏和肠系膜淋巴结(MLN)中最为丰富,而ASC主要在肝脏、脾脏和肾脏中表达。在心肌细胞、肝细胞、局灶性淋巴细胞、细支气管上皮细胞、肾小管上皮细胞、皮质神经元和MLN生发中心的内皮细胞中检测到NLRP3、ASC和半胱天冬酶-1的组织学分布。这些结果将有助于进一步研究NLRP3炎性小体的功能,并阐明其在山羊炎症性疾病中的作用。