CLONING, HETEROLOGOUS EXPRESSION AND DEVELOPMENTAL REGULATION OF A DROSOPHILA RECEPTOR FOR TACHYKININ-LIKE PEPTIDES
CLONING, HETEROLOGOUS EXPRESSION AND DEVELOPMENTAL REGULATION OF A DROSOPHILA RECEPTOR FOR TACHYKININ-LIKE PEPTIDES
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DOI：
10.1002/j.1460-2075.1991.tb04885.x
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发表时间：
1991-11-01
期刊：
影响因子：
11.4
通讯作者：
FORTE, M
中科院分区：
文献类型：
--
作者：
LI, XJ;WOLFGANG, W;FORTE, M
我们确定了克隆编码果蝇受体速激肽样肽的胚胎cDNA文库与探针从牛K物质受体的低严格筛选。该cDNA编码一个519个氨基酸的七跨膜结构域蛋白（DTKR），与哺乳动物速激肽受体的跨膜区有40 - 48%的氨基酸同一性。注射DTKR cRNA的非洲爪蟾卵母细胞对脊椎动物P物质及其激动剂表现出选择性反应，而对其他脊椎动物速激肽肽没有反应。用百日咳毒素处理卵母细胞可消除这些反应。在成年苍蝇，北方和PCR分析表明DTKR的优先表达的头部，原位杂交表明，DTKR是积累在成年中枢神经系统的神经元的细胞体。DTKR转录水平在发育过程中受到调节。北方和PCR扩增分析表明，虽然DTKR转录本存在于所有阶段，高水平的表达发生在胚胎发育的后期阶段（开始于10 - 14小时），与神经发育的主要时期的开始相一致。全胚胎原位杂交表明，DTKR表达在胚胎发育的后期阶段（11 - 15小时）在大脑中，并在一个特定的子集的神经元在每个neuromere的发展腹侧神经节。通过原位杂交将编码DTKR的基因定位到3号染色体右臂上99D的单个位置。这些观察结果表明，速激肽家族的肽递质及其受体代表了神经系统内细胞通讯的一种进化上古老的形式。此外，编码DTKR的转录本的丰度及其精确的发育调节表明，速激肽及其受体可能对正常发育至关重要。
We identified clones encoding a Drosophila receptor for tachykinin-like peptides by low stringency screening of an embryonic cDNA library with probes from the bovine substance K receptor. The cDNAs encode a seven transmembrane domain protein (DTKR) of 519 amino acids with 40-48% amino acid identity to mammalian tachykinin receptors within transmembrane regions. Xenopus oocytes injected with DTKR cRNAs showed selective responses to vertebrate substance P, its agonists and not to other vertebrate tachykinin peptides. These responses were eliminated by treatment of oocytes with pertussis toxin. In the adult fly, Northern and PCR analysis demonstrated preferential expression of DTKR in the head; in situ hybridization indicated that DTKR is accumulated in the cell bodies of neurons in the adult CNS. The levels of DTKR transcript are regulated during development. Northern and PCR amplification analysis showed that while DTKR transcripts are present at all stages, high levels of expression occur in later stages of embryogenesis (starting at 10 - 14 h), coinciding with the beginning of major periods of neural development. Whole mount embryo in situ hybridization demonstrated that DTKR is expressed at these later stages of embryogenesis (11-15 h) in the brain and in a specific subset of neurons in each neuromere of the developing ventral ganglion. The gene encoding DTKR was mapped by in situ hybridization to a single location at 99D on the right arm of chromosome 3. These observations demonstrate that the tachykinin family of peptide transmitters and their receptors represent an evolutionarily ancient form of cellular communication within the nervous system. In addition, the abundance of transcripts coding for DTKR and their precise developmental regulation suggest that tachykinins and their receptors may be critical for normal development.