Molecular cloning, expression and identification of the promoter regulatory region for the neuropeptide trissin in the nervous system of the silkmoth Bombyx mori
Molecular cloning, expression and identification of the promoter regulatory region for the neuropeptide trissin in the nervous system of the silkmoth Bombyx mori
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DOI：
10.1007/s00441-015-2352-z
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发表时间：
2016-06-01
影响因子：
3.6
通讯作者：
Zitnan, Dusan
中科院分区：
文献类型：
--
作者：
Roller, Ladislav;Cizmar, Daniel;Zitnan, Dusan
Trissin是一种保守的昆虫神经肽，但其在细胞中的表达和功能尚不清楚。我们检测到这种神经肽的存在，在家蚕的电子搜索和分子克隆。采用原位杂交技术研究了Trissin在幼虫、蛹和成虫整个中枢神经系统（CNS）和肠道中的表达。令人惊讶的是，它的表达仅限于两对小的前脑中间神经元和四到五个大的神经元在额神经节（FG）。这些神经元的进一步特征在于随后的多重染色与选定的抗体对昆虫神经肽。脑中间神经元支配蘑菇体的边缘，并与肌抑制肽（MIP）和CRF样利尿激素（CRF-DH）共表达Trissin。在FG中，一对神经元共表达Trissin、降钙素样利尿激素（CT-DH）、短神经肽F（sNPF）和MIP。这些神经元支配大脑、后脑和前中肠的肌肉组织。FG中的另一对trissin神经元共表达sNPF，并将轴突投射到tritcerebrum和中肠。我们还使用杆状病毒表达系统，以确定在这些神经元中的各种分子标记物的靶向表达的trissin基因的启动子调控区。Trissin在FG中的优势表达表明其可能在调节前肠-中肠收缩和食物摄入中起作用。
Trissin has recently been identified as a conserved insect neuropeptide, but its cellular expression and function is unknown. We detected the presence of this neuropeptide in the silkworm Bombyx mori using in silico search and molecular cloning. In situ hybridisation was used to examine trissin expression in the entire central nervous system (CNS) and gut of larvae, pupae and adults. Surprisingly, its expression is restricted to only two pairs of small protocerebral interneurons and four to five large neurons in the frontal ganglion (FG). These neurons were further characterised by subsequent multiple staining with selected antibodies against insect neuropeptides. The brain interneurons innervate edges of the mushroom bodies and co-express trissin with myoinhibitory peptides (MIP) and CRF-like diuretic hormones (CRF-DH). In the FG, one pair of neurons co-express trissin with calcitonin-like diuretic hormone (CT-DH), short neuropeptide F (sNPF) and MIP. These neurons innervate the brain tritocerebrum and musculature of the anterior midgut. The other pair of trissin neurons in the FG co-express sNPF and project axons to the tritocerebrum and midgut. We also used the baculovirus expression system to identify the promoter regulatory region of the trissin gene for targeted expression of various molecular markers in these neurons. Dominant expression of trissin in the FG indicates its possible role in the regulation of foregut-midgut contractions and food intake.