Analysis of species-specific function of neuropeptides in insects.

1.应用抗psps单克隆抗体，研究了psps相关肽在鳞翅目昆虫中枢神经系统中的分布。在家蚕和其他鳞翅目昆虫的大脑和心脏中，PRPs的不同分子似乎占主导地位。甘薯角虫（Agrius convoluvulli）在蜕皮期似乎从表直肠腺释放PRPs，但这些昆虫从该腺体分泌的时间不同。这些结果表明，鳞翅目昆虫中PRP的分布和分泌是不同的。从家蚕CNS中克隆了corazonin （Crz）受体的cDNA。实时荧光定量PCR分析表明，该受体主要在上腺、前丝腺和中枢神经系统中表达，而在表皮中不表达，尽管表皮被认为是蝗虫corazonin的可能靶组织。这一结果表明，corazonin受体的分布在不同的昆虫种类中是不同的，从而导致了不同的生理功能。Crz基因是从工蜂（Apis mellifera）的大脑中克隆出来的。ApimeCrz cDNA编码一种新的corazonin，显示[His^7]-corazonin的Thr^4取代。从家蚕的大脑和前胸神经节中分离到新的前胸抑制因子Bombyx myosuppressin （BMS）和Bombyx FMRFamide (BRFa)，分别克隆了编码这些肽的cdna。直接质谱分析和电生理分析表明，BRFa在5龄早期至中期通过前胸神经节神经支配的神经元供应到前胸腺（PG）表面。据我们所知，这项研究首次报道了肽通过直接神经支配控制卵巢甾体生成。

1.Distribution of prothoracicostatic peptide (PTSP)-related peptides (PRPs) in the central nervous system (CNS) was investigated using the monoclonal antibodies against PTSP in several lepidopteran insects. In the brain and corpora-cardiaca, different molecules of PRPs seemed to be predominant between Bombyx mori and other lepidopteran insects. In the sweet potato hornworm, Agrius convoluvulli, PRPs seemed to be released from the epiproctodeal glands during molting period, but the timing of secretion from this gland was different among these insects. These results suggest that the distribution and secretion of PRP are different among the lepidopteran insects.2.The cDNA encoding the receptor for corazonin (Crz) was cloned from the CNS of B.mori. Quantitative real-time PCR analysis suggested that this receptor was predominantly expressed hi the epitracheal gland, anterior silkgland, and CNS, but not in the epidermis, although epidermis is thought to be a possible target tissue of corazonin in the locusta. This result suggests that the distribution of corazonin receptor is different among insect species, which causes the different physiological function. The Crz gene was cloned from brains of the honey bee workers (Apis mellifera). The ApimeCrz cDNA encodes a new form of corazonin displaying Thr^4 substitution of [His^7]-corazonin.3.New prothoracicostatic factors, Bombyx myosuppressin (BMS) and Bombyx FMRFamide (BRFa), were isolated and the cDNAs encoding these peptides cloned from the brains and prothoracic ganglion of B.mori, respectively. Direct mass spectrometory and electrophysiological analyses demonstrated that BRFa was supplied to the surface of the prothoracic gland (PG) via neurons innervating from the prothoracic ganglion during early to mid fifth instar. To our knowledge, this study provides the first report of peptides controlling ecdysteroidogenesis by direct innervation.