Objective: To investigate the expression of some genes in Pomacea canaliculata infected with Angiostrongylus cantonensis, so as to provide insight into the preliminary understanding of the interactions between Angiostrongylus cantonensis and its intermediate host Pomacea canaliculata.
Methods: P. canaliculata was fed with rat faces containing the first-stage larvae of A. cantonensis. Three to five P. canaliculata was sampled 1, 10 days and 20 days after feeding, and the hemolymph, hepatopancreas, kidney, intestinal tract, head-foot and gill tissues were collected, while uninfected P. canaliculata served as controls. Total RNA was extracted from various tissues of P. canaliculata at different time points post-infection, and transcribed reversely into cDNA. Based on previous transcriptome sequencing results, 10 genes associated with immune defense, signal transduction, cell growth and metabolism, stress response were selected, and the gene expression was determined in the hemolymph tissues of P. canaliculata 1, 10 days and 20 days post-infection with A. cantonensis using real-time fluorescent quantitative PCR assay, and the α-tubulin gene expression was quantified in the hepatopancreas, kidney, head-foot, intestinal tract and gill tissues of P. canaliculata infected with A. cantonensis.
Results: Higher CELA1 gene expression was detected in the infection group than in the control group 1 (t = 12.32, P < 0.05), 10 days (t = 23.51, P < 0.05) and 20 days post-infection (t = 34.92, P < 0.05), and the CELA1 expression increased with the time of infection. The GST gene expression was (7.26 ± 1.80) times higher in the infection group than in the control group 1 day post-infection, and was significantly lower in the infection group than in the control group 10 days (t = 23.89, P < 0.05) and 20 days post-infection (t = 19.83, P < 0.05). Higher ferritin gene expression was found in the infection group than in the control group 10 days post-infection (t = 32.76, P < 0.05), and higher CRT gene expression was seen in the infection group than in the control group 1 (t = 7.23, P < 0.05), 10 days (t = 5.78, P < 0.05) and 20 days post-infection (t = 6.32, P < 0.05). In addition, the greatest α-tubulin gene expression was observed in the the hepatopancreatic tissues of P. canaliculata (F = 17.58, P < 0.05), and the α-tubulin gene expression altered in various tissues of P. canaliculata post-infection with A. cantonensis, with the most remarkable reduction of α - tubulin gene expression seen in the hepatopancreatic tissues (P < 0.05).
Conclusions: Following A. cantonensis infection in P. canaliculata, the expression of multiple genes is altered, and the expression of α-tubulin gene is inhibited in multiple tissues. The findings provide a basis for the further elucidation of the interactions between P. canaliculata and A. cantonensis.
[摘要] 目的 探究小管福寿螺感染广州管圆线虫后部分基因表达情况, 初步了解广州管圆线虫与其中间宿主小管福 寿螺之间的相互作用关系, 为防治广州管圆线虫病提供基础数据。方法 取含有广州管圆线虫Ⅰ期幼虫的大鼠粪便喂 食福寿螺, 分别于感染后1、10、20 d各取3～5只小管福寿螺, 采集其血淋巴、肝胰腺、肾脏、肠道、头足和鳃组织, 以未感 染的小管福寿螺为空白对照组, 提取不同感染时期小管福寿螺各组织总RNA, 逆转录为cDNA。基于前期转录组测序结 果, 挑选10个涉及免疫防御、信号转导、细胞生长和代谢、应激反应等方面的基因, 对小管福寿螺感染广州管圆线虫1、10、20 d的血淋巴进行基因荧光定量表达分析, 并对α-微管蛋白 (α-tubulin) 基因在感染广州管圆线虫后的福寿螺肝胰 腺、肾脏、头足部、肠道、鳃组织中的表达水平进行分析。结果 与空白对照组相比, CELA1 基因在感染广州管圆线虫后 1、10、20 d的福寿螺中表达量上调 (t = 12.32、23.51、34.92, P 均< 0.05), 且表达量随感染时间延长而上升; GST 基因表达 量在感染后1 d为空白对照组的 (7.26 ± 1.80) 倍, 在感染后10、20 d表达水平低于空白对照组 (t = 23.89、19.83, P 均< 0.05); ferritin 基因在感染后10 d较空白对照组显著上调 (t = 32.76, P < 0.05)。CRT 基因表达水平在感染后1、10、20 d均 较空白对照组上调 (t = 7.23、5.78、6.32, P 均< 0.05)。α-tubulin 基因在小管福寿螺肝胰腺组织中的表达水平最高 (F = 17.58, P < 0.05); 在小管福寿螺感染广州管圆线虫后, 该基因在各组织中的表达量均有不同程度变化, 其表达量以肝胰 腺组织中下调最为明显 (P均< 0.05)。结论 小管福寿螺感染广州管圆线虫后, 血淋巴中多个基因表达水平发生改变, α-tubulin基因表达水平在多个组织中受到抑制。该研究为深入阐明小管福寿螺和广州管圆线虫的相互作用关系奠定了基础。.
Keywords: Angiostrongylus cantonensis; Interaction; Pomacea canaliculata; α-tubulin.