敲低piwi基因对黑腹果蝇血细胞增殖及分化的影响  

Effect of piwi knock-down on hemocyte proliferation and differentiation in Drosophila melanogaster

作　　者：刘晓楠 赵素娟 王博 王宏鑫 郝阳光 LIU Xiao-Nan;ZHAO Su-Juan;WANG Bo;WANG Hong-Xin;HAO Yang-Guang(Department of Basic Medical,Shenyang Medical College,Shenyang 110034,China)

机构地区：[1]沈阳医学院基础医学院,沈阳110034

出　　处：《昆虫学报》2021年第12期1359-1366,共8页Acta Entomologica Sinica

基　　金：国家自然科学基金青年科学基金项目(31900366);辽宁省自然科学基金项目(20180530021);辽宁省大学生创新创业科研基金项目(20198019);沈阳医学院博士科研启动基金项目(20186064)。

摘　　要：【目的】探究敲低piwi基因对黑腹果蝇Drosophila melanogaster血细胞增殖及分化的影响。【方法】利用黑腹果蝇e33C-Gal4和Hml-Gal4-UAS-2×EGFP品系分别与野生型w^(1118)和UAS-piwi RNAi品系杂交,实现在黑腹果蝇游离血细胞或淋巴腺中降低piwi基因的表达;采用免疫荧光染色方法检测Piwi蛋白在血细胞中的定位及其对黑腹果蝇血细胞增殖与分化的影响。【结果】Piwi蛋白在黑腹果蝇游离血细胞及整个淋巴腺中都表达,且主要定位在细胞质;敲低piwi基因导致游离血细胞数量明显增加,处于有丝分裂M期的细胞数量增加,但未影响游离血细胞中浆细胞及薄层细胞的分化;敲低piwi基因对淋巴腺血细胞增殖无影响,但导致浆细胞过度分化及薄层细胞的产生。【结论】piwi基因在果蝇游离血细胞中的缺失可引起血细胞过度增殖,而在淋巴腺中敲低可引起血细胞的异常分化。【Aim】To explore the effect of piwi knock-down on hemocyte proliferation and differentiation in Drosophila melanogaster.【Methods】The D.melanogaster strains e33C-Gal4 and Hml-Gal4-UAS-2×EGFP were crossed with the wild strain w^(1118) and the strain UAS-piwi RNAi,respectively,to reduce the expression of piwi gene in circulating hemocytes or lymph glands of D.melanogaster.The localization of Piwi protein in hemocytes and its effect on hemocyte proliferation and differentiation in D.melanogaster were detected by immunofluorescence staining.【Results】Piwi protein was expressed in circulating hemocytes and the whole lymph glands of D.melanogaster,and was mainly localized in the cytoplasm.The knock-down of piwi gene resulted in significant increase in the number of circulating hemocytes and the number of cells in the M phase of mitosis,but the differentiation of plasmatocytes and lamellocytes in circulating hemocytes were not affected.The knock-down of piwi gene had no effect on the proliferation of lymph gland hemocytes,but resulted in the excessive differentiation of plasmatocytes and the generation of lamellocytes.【Conclusion】The knock-down of piwi gene in circulating hemocytes in Drosophila can induce the overproliferation of hemocytes,while the knock-down of piwi gene in lymph glands can induce abnormal differentiation of hemocytes.

关 键 词：黑腹果蝇 PIWI 淋巴腺 血细胞 增殖与分化 免疫荧光染色 

分 类 号：Q352[生物学—遗传学]