High-Level Expression, Purification, and Characterization of the Recombinant Grass Carp Pituitary Adenylate Cyclase-Activating Polypeptide

Wang, Zhangzhi; Mu, Yang; Wang, Xinyan; Zhou, Hong

doi:10.1271/bbb.80057

Cited by 5 publications

(1 citation statement)

References 30 publications

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“…Recently, it has been reported that the growth rates of catfish (Clarias gariepinus), tilapia (Oreochromis niloticus), and carp (Cyprinus carpio) were enhanced by the addition of both catfish PACAP-related peptide (PRP) and PACAP recombinant peptides (Lugo et al 2008). However, recombinant grass carp peptide specifically inhibited T98G human glioblastoma cell proliferation (Wang et al 2008). These results suggest that the growth-promoting effect of PACAP may be species-specific.…”

Section: Introductionmentioning

confidence: 99%

Expression and in vitro regulation of pituitary adenylate cyclase-activating polypeptide (pacap38) and its type I receptor (pac1-r) in the gonads of tilapia (Oreochromis mossambicus)

Huang¹,

Li²,

Wu³

et al. 2009

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Pituitary adenylate cyclase-activating polypeptide (PACAP), a pleiotropic neuropeptide, has diverse functions in mammals. However, studies of the expression and function of PACAP and its receptor in fish, particularly in the reproductive system, are still limited. In this report, semi-quantitative RT-PCR and immunohistochemical staining were performed to identify expression domains of commercially important tilapia (Oreochromis mossambicus). PACAP (tpacap 38 ) and its type I receptor (tpac 1 -r). Transcripts were detected in the brain, gallbladder, gill, heart, intestine, kidney, muscles, pancreas, spleen, stomach, testes, and ovaries, but not in the liver. Expression of tpacap 38 and tpac 1 -r mRNA in brain tissue was significantly higher in both sexes compared with other tissues. Addition of exogenous ovine PACAP 38 (0.25-5 nM), cAMP analog (dibutyryl-cAMP, 0.25-1.5 mM) or forskolin (adenylate cyclase activator, 1-10 mM) significantly upregulated tpacap 38 in the gonads via a dose-and time-dependent fashion. This effect reached a maximal level at 2 h after induction, and then decreased with prolonged culture for up to 4 or 8 h. Additionally, the expression levels of tpac 1 -r were not significantly affected by ovine PACAP 38 or dibutyryl-cAMP in either sex. Forskolin had a slightly inductive effect and its function could be suppressed with the addition of protein kinase A (PKA) inhibitor, H89 (10 mM), indicating involvement of the cAMP-PKA signaling pathway in the regulation of tpacap 38 . Expression of tpacap 38 and tpac 1 -r in the gonads of tilapia suggests that PACAP may mediate gonadotropin action via paracrine/autocrine mechanisms in this bony fish.

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Section: Introductionmentioning

confidence: 99%