Department of Biology, University of Toronto Mississauga, 3359 Mississauga Road North, Mississauga, Ontario, Canada L5L 1C6.
Gen Comp Endocrinol. 2011 Oct 1;174(1):36-43. doi: 10.1016/j.ygcen.2011.08.007. Epub 2011 Aug 22.
Crustacean cardioactive peptide (CCAP), a cyclic nonapeptide (PFCNAFTGCamide), has multifunctional roles in insects including stimulating visceral and cardiac muscle contraction, and regulating ecdysis. Previously, we have sequenced the cDNA for CCAP from Rhodnius prolixus central nervous system (CNS) and shown expression of the CCAP transcript in neurons of the CNS. In the present study, we have biochemically identified and sequenced CCAP from 5th instar R. prolixus CNS using matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry, and mapped CCAP-like immunoreactivity in the CNS and peripheral tissues of 5th instar R. prolixus. Physiologically, the hindgut of R. prolixus was found to be sensitive to CCAP, showing dose-dependent increases in contractions with threshold at 5 × 10(-9) M and maximum response at 10(-7) M CCAP. Also, CCAP was found to increase the frequency of the heartbeat in a reversible, dose-dependent manner, with threshold close to 10(-11) M and maximum response at 10(-10) M CCAP.
甲壳动物心脏活性肽(CCAP),一种环状九肽(PFCNAFTGCamide),在昆虫中具有多种功能,包括刺激内脏和心肌收缩,以及调节蜕皮。先前,我们已经从 Rhodnius prolixus 中枢神经系统(CNS)中测序了 CCAP 的 cDNA,并显示了 CCAP 转录物在 CNS 神经元中的表达。在本研究中,我们使用基质辅助激光解吸电离-飞行时间串联质谱法从 5 龄期 Rhodnius prolixus CNS 中生化鉴定和测序了 CCAP,并在 5 龄期 Rhodnius prolixus CNS 和外周组织中绘制了 CCAP 样免疫反应。生理学上,发现 Rhodnius prolixus 的后肠对 CCAP 敏感,在 5×10(-9) M 时呈剂量依赖性收缩增加,在 10(-7) M CCAP 时达到最大反应。此外,CCAP 以可逆、剂量依赖性的方式增加心跳频率,其阈值接近 10(-11) M,在 10(-10) M CCAP 时达到最大反应。