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家蚕脑视蛋白(Boceropsin)的分子克隆及其在蚕脑内表达的细胞定位

Molecular cloning of Bombyx cerebral opsin (Boceropsin) and cellular localization of its expression in the silkworm brain.

作者信息

Shimizu I, Yamakawa Y, Shimazaki Y, Iwasa T

机构信息

Division of Molecular Ecology, Center for Ecological Research, Kyoto University, Kamitanakami, Otsu, Shiga 522-2113, Japan.

出版信息

Biochem Biophys Res Commun. 2001 Sep 14;287(1):27-34. doi: 10.1006/bbrc.2001.5540.

Abstract

We have cloned a cDNA for a novel opsin from the larval brain of the silkworm Bombyx mori in which the photoperiodic photoreceptor had been supposed to reside in the cephalic central nervous system (CNS). Its deduced amino acid sequence was composed of 381 amino acids and included amino acid residues highly conserved in insect visual pigments. This opsin belonged to the long wavelength photoreceptor group of insect opsins and showed the greatest degree of homology (84%) with the green visual photoreceptor in the sphingid moth. We have designated this Bombyx cerebral opsin as Boceropsin. Southern blotting experiments indicated that the Boceropsin gene is present in a single copy, and RT-PCR analysis revealed that Boceropsin mRNA is expressed in the larval brain but not in the subesophageal ganglion (Sg) or thoracic ganglion (Tg). Immunohistochemical analyses demonstrated that Boceropsin protein is present bilaterally in some defined cells localized in the brain of Bombyx larvae. This is the first report of expression of an opsin-based protein in CNS of an insect. The possibility that the Boceropsin functions as the photoperiodic receptive pigment in the silkworm is also discussed.

摘要

我们从家蚕幼虫大脑中克隆了一种新型视蛋白的cDNA,此前认为光周期光感受器存在于家蚕头部中枢神经系统(CNS)中。其推导的氨基酸序列由381个氨基酸组成,包含在昆虫视觉色素中高度保守的氨基酸残基。这种视蛋白属于昆虫视蛋白的长波长光感受器组,与天蛾科蛾类的绿色视觉光感受器具有最高程度的同源性(84%)。我们将这种家蚕脑视蛋白命名为Boceropsin。Southern印迹实验表明Boceropsin基因以单拷贝形式存在,RT-PCR分析显示Boceropsin mRNA在幼虫大脑中表达,但在咽下神经节(Sg)或胸神经节(Tg)中不表达。免疫组织化学分析表明,Boceropsin蛋白在位于家蚕幼虫大脑中的一些特定细胞中双侧存在。这是首次报道昆虫中枢神经系统中基于视蛋白的蛋白质表达。还讨论了Boceropsin在家蚕中作为光周期感受色素发挥作用的可能性。

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