Choi Cheol Young, Zheng Junying, Watson R Douglas
Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Gen Comp Endocrinol. 2006 Sep 15;148(3):383-7. doi: 10.1016/j.ygcen.2006.03.003. Epub 2006 May 2.
Crustacean hyperglycemic hormone (CHH), a polypeptide with multiple physiological effects, was first identified in the X-organ/sinus gland neurosecretory system of the eyestalks. In studies reported here, we used a PCR-based cloning strategy (RT-PCR followed by 5'- and 3'-RACE) to clone from blue crab (Callinectes sapidus) eyestalk ganglia a cDNA (CsCHH-1) encoding a putative CHH preprohormone. Sequence analysis revealed the preprohormone included all structural features previously reported for CHH preprohormones: a signal peptide, a CHH precursor-related peptide (CPRP), the CHH polypeptide, and a C-terminal basic processing site. Further, the deduced amino acid sequence of the mature polypeptide included all signature domains previously reported for CHH. The primary structure of blue crab CHH is most closely related to CHH from other brachyurans. RT-PCR revealed the CsCHH-1 transcript was present in eyestalk ganglia, but was undetectable in other tissues tested. A transcript encoding a similar CHH-like preprohormone was detected in thoracic ganglion, ventral nerve cord, and brain, but was not detected in eyestalk ganglia.
甲壳类高血糖激素(CHH)是一种具有多种生理效应的多肽,最初是在眼柄的X器官/窦腺神经分泌系统中发现的。在本文报道的研究中,我们采用基于PCR的克隆策略(逆转录聚合酶链反应(RT-PCR),随后进行5'和3'端快速扩增(RACE))从蓝蟹(Callinectes sapidus)眼柄神经节中克隆出一个编码假定CHH前激素原的cDNA(CsCHH-1)。序列分析表明,该前激素原包含先前报道的CHH前激素原的所有结构特征:一个信号肽、一个CHH前体相关肽(CPRP)、CHH多肽以及一个C端碱性加工位点。此外,推导的成熟多肽氨基酸序列包含先前报道的CHH的所有特征结构域。蓝蟹CHH的一级结构与其他短尾类动物的CHH最为密切相关。RT-PCR显示CsCHH-1转录本存在于眼柄神经节中,但在其他测试组织中未检测到。在胸神经节、腹神经索和脑中检测到一个编码类似CHH样前激素原的转录本,但在眼柄神经节中未检测到。