Guo Cheng, Xie Yue, Liu Yuchen, Wang Ning, Zhan Jiafei, Zhou Xuan, Angel Christiana, Gu Xiaobin, Lai Weimin, Peng Xuerong, Yang Guangyou
Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.
Genes (Basel). 2018 Nov 19;9(11):559. doi: 10.3390/genes9110559.
, the metacestode of , causes coenurosis, a disease severely affecting goat, sheep, cattle and yak farming and resulting in huge economic losses annually. Annexins bind calcium ions and play an important role in flatworm parasite development. To explore potential functions of annexins in , three homologous genes, namely, , and , were screened from the transcriptome dataset, amplified from cDNA and subjected to bioinformatics analysis. Then, polyclonal antibodies recognizing the recombinant TmAnxB2 (rTmAnxB2) and rTmAnxB3 were prepared for localization of TmAnxB2 and TmAnxB3 in different tissues and developmental stages by immunofluorescence. The transcription of all three genes was also measured by relative fluorescent quantitative PCR. The sizes of rTmAnxB2, rTmAnxB3 and rTmAnxB12 were 58.00, 53.06 and 53.51 kDa, respectively, and rTmAnxB12 was unstable. Both rTmAnxB2 and rTmAnxB3 were recognized by goat-positive sera in Western blots. Immunofluorescence revealed that TmAnxB2 and TmAnxB3 were localized in the protoscolex and cyst wall and TmAnxB3 was also detected in adult cortex. TmAnxB2 and TmAnxB12 mRNA levels were determined to be highest in oncospheres and protoscolex, whereas transcription of TmAnxB3 was highest in scolex and immature segments. Taken together, these findings indicate that TmAnxB2 and TmAnxB12 may play critical roles in larvae, while TmAnxB3 may have important functions in adults. These results will lay the foundation for functional research of annexins in .
,的中绦期会引发多头蚴病,这是一种严重影响山羊、绵羊、牛和牦牛养殖的疾病,每年都会造成巨大的经济损失。膜联蛋白结合钙离子并在扁形寄生虫发育中发挥重要作用。为了探究膜联蛋白在中的潜在功能,从转录组数据集中筛选出三个同源基因,即、和,从的cDNA中扩增出来并进行生物信息学分析。然后,制备了识别重组TmAnxB2(rTmAnxB2)和rTmAnxB3的多克隆抗体,通过免疫荧光法对TmAnxB2和TmAnxB3在不同组织和发育阶段进行定位。还通过相对荧光定量PCR测定了这三个基因的转录情况。rTmAnxB2、rTmAnxB3和rTmAnxB12的大小分别为58.00、53.06和53.51 kDa,且rTmAnxB12不稳定。在蛋白质免疫印迹中,山羊阳性血清可识别rTmAnxB2和rTmAnxB3。免疫荧光显示,TmAnxB2和TmAnxB3定位于原头节和囊壁,在成虫皮层中也检测到了TmAnxB3。测定发现TmAnxB2和TmAnxB12的mRNA水平在六钩蚴和原头节中最高,而TmAnxB3的转录在头节和未成熟节片中最高。综上所述,这些发现表明TmAnxB2和TmAnxB12可能在幼虫中起关键作用,而TmAnxB3可能在成虫中具有重要功能。这些结果将为膜联蛋白在中的功能研究奠定基础。