D'Aquila Andrea L, Hsieh Alan Hwa-Ruey, Hsieh Adam Hwa-Ming, De Almeida Reuben, Lovejoy Sabine R, Lovejoy David A
Department of Cell and Systems Biology, University of Toronto, Ontario, Canada.
Department of Cell and Systems Biology, University of Toronto, Ontario, Canada.
Gen Comp Endocrinol. 2017 May 15;246:105-115. doi: 10.1016/j.ygcen.2016.06.015. Epub 2016 Jun 9.
Teneurin C-terminal associated peptide (TCAP) is a neuropeptide that bears some structural similarity to the corticotropin-releasing factor (CRF) family of peptides. TCAP and CRF are both implicated in the regulation of stress-related behaviors, as established in rodent models. However, in vertebrates, both TCAP and CRF possess three additional paralogous forms making vertebrate models difficult to assess with respect to TCAP-CRF interaction. As a urochordate, this species possesses single homologs of TCAP and of a CRF/Diuretic-like peptide (CDLP) in the genome, thereby establishing Ciona intestinalis as an excellent model organism to examine the interaction of these peptide systems. However, the lack of C. intestinalis synthetic peptides and specific antisera has complicated experimentation. We, therefore, prepared synthetic versions of CDLP and TCAP to prepare specific antisera and to investigate their bioactivity in this species. To analyze stress-related behaviors, a novel behavioral assay was used to characterize different types of contraction-based behaviors, using buccal opening contractions, cloacal opening contractions, lateral contractions, longitudinal contractions and expulsions. Protein and mRNA expression data indicate that the mature versions of both peptides are present in a number of tissues. With respect to behavioral activity, both TCAP- and CDLP-treated animals had distinct contraction profiles under ambient conditions. Moreover, food stimulation tests revealed that whereas CDLP-treated animals displayed a strong expulsion behavior in response to feeding, TCAP-treated animals did not show this effect. These actions are consistent with previous studies done in vertebrates.
Tenascin C末端相关肽(TCAP)是一种神经肽,与促肾上腺皮质激素释放因子(CRF)家族的肽在结构上有一些相似之处。如在啮齿动物模型中所证实的,TCAP和CRF都与应激相关行为的调节有关。然而,在脊椎动物中,TCAP和CRF都有另外三种旁系同源形式,这使得脊椎动物模型难以评估TCAP与CRF的相互作用。作为一种尾索动物,该物种在基因组中拥有TCAP和CRF/利尿样肽(CDLP)的单一同源物,从而使海鞘成为研究这些肽系统相互作用的优秀模式生物。然而,缺乏海鞘合成肽和特异性抗血清使实验变得复杂。因此,我们制备了CDLP和TCAP的合成版本,以制备特异性抗血清并研究它们在该物种中的生物活性。为了分析应激相关行为,我们使用了一种新颖的行为测定方法,通过口腔开口收缩、泄殖腔开口收缩、侧向收缩、纵向收缩和排出等基于收缩的行为来表征不同类型的行为。蛋白质和mRNA表达数据表明,这两种肽的成熟版本都存在于许多组织中。关于行为活性,在环境条件下,经TCAP和CDLP处理的动物都有不同的收缩模式。此外,食物刺激试验表明,经CDLP处理的动物在进食时表现出强烈的排出行为,而经TCAP处理的动物则没有这种效应。这些作用与先前在脊椎动物中进行的研究一致。