Winther Asa M E, Siviter Richard J, Isaac R Elwyn, Predel Reinhard, Nässel Dick R
Department of Zoology, Stockholm University, SE-10691 Stockholm, Sweden.
J Comp Neurol. 2003 Sep 15;464(2):180-96. doi: 10.1002/cne.10790.
The gene Dtk, encoding the prohormone of tachykinin-related peptides (TRPs), has been identified from Drosophila. This gene encodes five putative tachykinin-related peptides (DTK-1 to 5) that share the C-terminal sequence FXGXRamide (where X represents variable residues) as well as an extended peptide (DTK-6) with the C-terminus FVAVRamide). By mass spectrometry (MALDI-TOF-MS), we identified ion signals with masses identical to those of DTK-1 to 5 in specific brain regions. We have analyzed the distribution of the Dtk transcript and peptides, by in situ hybridization and immunocytochemistry during postembryonic development of the central nervous system (CNS) of Drosophila. Antiserum against a cockroach TRP that cross-reacts with the DTKs was used for immunocytochemistry. Expression of transcript and peptides was detected from first to third instar larvae, through metamorphosis to adult flies. Throughout postembryonic development, we were able to follow the strong expression of TRPs in a pair of large descending neurons with cell bodies in the brain. The number of TRP-expressing neuronal cell bodies in the brain and ventral nerve cord increases during larval development. In the early pupa (stage P8), the number of TRP-expressing cell bodies is lower than in the third instar larvae. The number drastically increases during later pupal development, and in the adult fly about 200 TRP-expressing neurons can be seen in the CNS. The continuous expression of TRPs in neurons throughout postembryonic development suggests specific functional roles in both larval and imaginal flies and possibly also in some neurons during pupal development.
已从果蝇中鉴定出编码速激肽相关肽(TRP)前体激素的基因Dtk。该基因编码五种假定的速激肽相关肽(DTK-1至5),它们共享C末端序列FXGXRamide(其中X代表可变残基)以及具有C末端FVAVRamide的延伸肽(DTK-6)。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS),我们在特定脑区鉴定出了质量与DTK-1至5相同的离子信号。我们通过原位杂交和免疫细胞化学分析了果蝇中枢神经系统(CNS)胚胎后发育过程中Dtk转录本和肽的分布。使用与DTK发生交叉反应的针对蟑螂TRP的抗血清进行免疫细胞化学分析。从一龄幼虫到三龄幼虫,直至变态发育为成虫,均检测到转录本和肽的表达。在整个胚胎后发育过程中,我们能够追踪到一对位于大脑中具有细胞体的大型下行神经元中TRP的强烈表达。在幼虫发育过程中,大脑和腹神经索中表达TRP的神经元细胞体数量增加。在蛹早期(P8期),表达TRP的细胞体数量低于三龄幼虫。在蛹后期发育过程中,数量急剧增加,在成虫中,中枢神经系统中可见约200个表达TRP的神经元。TRP在整个胚胎后发育过程中在神经元中的持续表达表明其在幼虫和成虫阶段以及可能在蛹发育过程中的某些神经元中具有特定的功能作用。