Owa Chie, Aoki Fugaku, Nagata Masao
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Seimei Building 302, 5-1-5 Kashiwa-no-ha, Kashiwa, Chiba 277-8562, Japan.
J Insect Physiol. 2008 Aug;54(8):1286-92. doi: 10.1016/j.jinsphys.2008.06.011. Epub 2008 Jul 16.
To examine the function of silkworm Bombyx mori L. athrocytes (nephrocytes), we constructed cDNAs of larval peritracheal athrocytes that were anatomically isolated from surrounding tissues. Larval expression levels of genes encoding hemolymph proteins, such as arylphorin, the 30K proteins, and lysozyme, were lower in peritracheal athrocytes than in the fat body, whereas genes involved in protein degradation were highly expressed in athrocytes. Real time RT-PCR revealed that a member of the Hsp40/Dnaj protein family, DjA2 (also known as Rdj2, Dj3, Dnj3, Cpr3, and Hirip4), an endocytic gene, was highly expressed in the peritracheal athrocytes compared to the fat body. Homologs of the Drosophila ATG1, ATG5, ATG6, and ATG8 genes had high expression levels in the peritracheal athrocytes. Observations using laser confocal microscopy with lysosomal fluorescent probes showed that silkworm athrocytes, including pericardial cells, suboesophageal body, and peritracheal athrocytes, were rich in lysosomes, in contrast to other tissues. Peritracheal athrocytes had lysotracker-positive spots at all times from the fourth larval molt to the pupa. Of these, molting larval and pupal peritracheal athrocytes had larger spots. Starvation for 24h induced greater lysotracker staining, but the number of spots decreased. Silkworm peritracheal athrocytes are lysosome-rich tissues and may function in the degradation of proteins.
为了研究家蚕血细胞(肾细胞)的功能,我们构建了从周围组织中解剖分离出的幼虫气管周血细胞的cDNA。编码血淋巴蛋白的基因,如芳基脂蛋白、30K蛋白和溶菌酶,在气管周血细胞中的幼虫表达水平低于脂肪体,而参与蛋白质降解的基因在血细胞中高度表达。实时RT-PCR显示,与脂肪体相比,如果蝇ATG1、ATG5、ATG6和ATG8基因的同源物在气管周血细胞中具有高表达水平。使用溶酶体荧光探针的激光共聚焦显微镜观察表明,与其他组织相比,家蚕血细胞,包括心包细胞、咽下神经节和气管周血细胞,富含溶酶体。从幼虫第四次蜕皮到蛹期,气管周血细胞始终有溶酶体示踪剂阳性斑点。其中,蜕皮幼虫和蛹期的气管周血细胞斑点较大。饥饿24小时会导致溶酶体示踪剂染色增强,但斑点数量减少。家蚕气管周血细胞是富含溶酶体的组织,可能在蛋白质降解中发挥作用。