Siekhaus D E, Fuller R S
Department of Biochemistry, Stanford University School of Medicine, Stanford, California 94305-5307, USA.
J Neurosci. 1999 Aug 15;19(16):6942-54. doi: 10.1523/JNEUROSCI.19-16-06942.1999.
Accurate proteolytic processing of neuropeptide and peptide hormone precursors by members of the kexin/furin family of proteases is key to determining both the identities and activities of signaling peptides. Here we identify amontillado (amon), the Drosophila melanogaster homolog of the mammalian neuropeptide processing protease PC2, and show that in contrast to vertebrate PC2, amontillado expression undergoes extensive regulation in the nervous system during development. In situ hybridization reveals that expression of amontillado is restricted to the final stages of embryogenesis when it is found in anterior sensory structures and in only 168 cells in the brain and ventral nerve cord. After larvae hatch from their egg shells, the sensory structures and most cells in the CNS turn off or substantially reduce amontillado expression, suggesting that amontillado plays a specific role late in embryogenesis. Larvae lacking the chromosomal region containing amontillado show no gross anatomical defects and respond to touch. However, such larvae show a greatly reduced frequency of a hatching behavior of wild-type Drosophila in which larvae swing their heads, scraping through the eggshell with their mouth hooks. Ubiquitous expression of amontillado can restore near wild-type levels of this behavior, whereas expression of amontillado with an alanine substitution for the catalytic histidine cannot. These results suggest that amontillado expression is regulated as part of a programmed modulation of neural signaling that controls hatching behavior by producing specific neuropeptides in particular neurons at an appropriate developmental time.
蛋白酶的克新/弗林家族成员对神经肽和肽激素前体进行精确的蛋白水解加工,是决定信号肽的身份和活性的关键。在这里,我们鉴定出了蒙蒂亚托(amon),它是哺乳动物神经肽加工蛋白酶PC2在果蝇中的同源物,并表明与脊椎动物的PC2不同,蒙蒂亚托的表达在发育过程中在神经系统中受到广泛调控。原位杂交显示,蒙蒂亚托的表达仅限于胚胎发育的最后阶段,此时它存在于前部感觉结构以及大脑和腹神经索中仅168个细胞中。幼虫从卵壳中孵化出来后,感觉结构和中枢神经系统中的大多数细胞关闭或大幅降低蒙蒂亚托的表达,这表明蒙蒂亚托在胚胎发育后期发挥特定作用。缺乏包含蒙蒂亚托的染色体区域的幼虫没有明显的解剖学缺陷,并且对触摸有反应。然而,这些幼虫表现出野生型果蝇孵化行为的频率大大降低,在这种行为中,幼虫摆动头部,用口钩刮破卵壳。蒙蒂亚托的普遍表达可以恢复这种行为接近野生型的水平,而用丙氨酸替代催化组氨酸的蒙蒂亚托的表达则不能。这些结果表明,蒙蒂亚托的表达作为神经信号程序性调节的一部分受到调控,该调节通过在适当的发育时间在特定神经元中产生特定神经肽来控制孵化行为。