Genetics & Developmental Biology Laboratory, School of Biological Sciences & Biotechnology, Indian Institute of Advanced Research/IAR, Koba Institutional Area, Gandhinagar, Gujarat, 382007, India.
Mol Neurobiol. 2018 May;55(5):4345-4361. doi: 10.1007/s12035-017-0650-z. Epub 2017 Jun 20.
In the present study, we demonstrate that molecular chaperone Hsp70 and Hsc70 is essential for normal organization and development of ommatidial cells in Drosophila melanogaster eye. An exogenously expressed dominant negative mutant of Hsp70 (K71E) and Hsc70.4 (K71S and D206S) in an eye-specific manner resulted in eye degeneration that includes loss of eye pigment, disorganized ommatidia, abnormality in bristle cell arrangement and reduction in the eye size. The developmental organization of ommatidial cells (cone, photoreceptor, pigment, and bristle cell complex) was disturbed in Hsp70 and Hsc70 mutants. Acridine orange (AO) and caspase 3 staining showed an increased cell death in Hsp70 and Hsc70 mutant eyes. Genetic interaction study of Hsp70 and Hsc70 mutants with candidate genes of JNK signaling pathway and immunocytochemistry study using phospho-JNK antibody suggested that mutation in Hsp70 and Hsc70 results in ectopic activation of JNK signaling in fly eye. Further, anti-PH3 staining in Hsp70 and Hsc70 mutant eyes revealed a reduced number of mitotic cells in second mitotic wave (SMW) of developing eye and anti-Rh1 staining showed reduced Rh1 expression, accumulation of Rh1 in the cytoplasm, and rhabdomere degeneration. Thus, on the basis of results, it was concluded that molecular chaperone Hsp70 and Hsc70 play an indispensable role during Drosophila eye development.
在本研究中,我们证明了分子伴侣 Hsp70 和 Hsc70 对于黑腹果蝇眼的小眼细胞的正常组织和发育是必不可少的。以眼睛特异性方式表达的 Hsp70(K71E)和 Hsc70.4(K71S 和 D206S)的显性负突变体导致眼睛退化,包括眼睛色素丧失、小眼排列紊乱、刚毛细胞排列异常以及眼睛缩小。小眼细胞(视锥细胞、感光细胞、色素和刚毛细胞复合物)的发育组织在 Hsp70 和 Hsc70 突变体中受到干扰。吖啶橙(AO)和 caspase 3 染色显示 Hsp70 和 Hsc70 突变体眼睛中的细胞死亡增加。Hsp70 和 Hsc70 突变体与 JNK 信号通路候选基因的遗传相互作用研究以及使用磷酸化-JNK 抗体的免疫细胞化学研究表明,Hsp70 和 Hsc70 的突变导致果蝇眼睛中 JNK 信号的异位激活。此外,在 Hsp70 和 Hsc70 突变体眼睛中的抗-PH3 染色显示发育中眼睛的第二次有丝分裂波(SMW)中的有丝分裂细胞数量减少,并且抗-Rh1 染色显示 Rh1 表达减少,Rh1 在细胞质中积累,并且光感受器退化。因此,根据结果得出结论,分子伴侣 Hsp70 和 Hsc70 在果蝇眼睛发育过程中发挥不可或缺的作用。