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果蝇中与序列相关的神经肽的免疫细胞化学

Immunocytochemistry of sequence-related neuropeptides in Drosophila.

作者信息

Tibbetts M F, Nichols R

机构信息

Department of Biology, University of Michigan, Ann Arbor 48109-1048.

出版信息

Neuropeptides. 1993 Jun;24(6):321-5. doi: 10.1016/0143-4179(93)90002-r.

Abstract

Based on structure, activity, and expression, the Drosophila drosulfakinin I peptide (DSK I; FDDY(OSO3H)GHMRFamide) is similar to the vertebrate peptide, cholecystokinin. Dromyosuppressin (DMS; TDVDHVFLRFamide) is an abundant peptide isolated from adult Drosophila which shares a high degree of sequence homology with peptides isolated from chicken, cockroach, fleshfly, and locust. DSK I and DMS, encoded by different precursors, have similar expression patterns in larval brain tissue; each localizes to cells in the anterior and medial protocerebrum. Because of the precedence for coexistence of neural messengers, it was of interest to determine the cellular expression patterns relative to one another. The question of whether the two peptides were expressed in the same cells was resolved using an immunofluorescent double-labeling technique developed for sequence-specific antisera raised in separate animals of the same species. Double labeling was done using a combination of indirect and direct immunofluorescence. DSK I and DMS were shown to localize to different cells in close proximity to one another in the larval brain. The non-overlapping expression patterns of these peptides illustrate the complete lack of cross-staining with this technique.

摘要

基于结构、活性和表达情况,果蝇的果蝇速激肽I肽(DSK I;FDDY(OSO3H)GHMRFamide)与脊椎动物的肽胆囊收缩素相似。促肌抑制素(DMS;TDVDHVFLRFamide)是从成年果蝇中分离出的一种丰富的肽,它与从鸡、蟑螂、麻蝇和蝗虫中分离出的肽具有高度的序列同源性。由不同前体编码的DSK I和DMS在幼虫脑组织中具有相似的表达模式;它们各自定位于前脑和中脑原脑的细胞中。由于神经信使共存的先例,确定它们彼此相对的细胞表达模式很有意义。使用为在同一物种的不同动物中产生的序列特异性抗血清开发的免疫荧光双标记技术,解决了这两种肽是否在同一细胞中表达的问题。双标记采用间接免疫荧光和直接免疫荧光相结合的方法进行。结果表明,在幼虫脑中,DSK I和DMS定位于彼此紧邻的不同细胞中。这些肽的非重叠表达模式表明,使用该技术完全不存在交叉染色。

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