Cloning and characterization of a PAC1 receptor hop-1 splice variant in goldfish (Carassius auratus)

Abstract : In several vertebrates, it has been demonstrated that alternative splicing of PAC1 receptor (PAC1-R) transcripts can generate a number of functional receptor variants which utilize diVerent signal transduction pathways to mediate their activities. As PACAP is a physiological growth hormone-releasing factor in Wsh, and PACAP and the PAC1-R are highly conserved in vertebrate evolution, it would be of interest to investigate the structure and cellular distribution, particularly in the pituitary, of PAC1-R splice variants in a Wsh model. Our laboratory has previously cloned a receptor cDNA corresponding to the goldWsh PAC1-R-s (goldWsh PAC1-R-short). In the present study, a goldWsh PAC1-R-hop1 variant was characterized. Functional expression of goldWsh PAC1-R-s and PAC1-R-hop1 in Chinese Hamster Ovary cells revealed that, upon stimulation by ovine PACAP38, these receptor variants exhibited similar EC 50 values (8.7 § 1.5 and 8.8 § 1.9 nM, respectively) and maximal responses in activating intracellular cAMP production. The presence and expression levels of these transcripts were measured by quantitative real-time PCR in the brain, heart, pituitary and male gonad, and goldWsh PAC1-R-s were found to be the predominant form. In situ hybridization of goldWsh PAC1-R in the pituitary revealed its prevalent presence in the pars distalis. In summary, the present study provides information to conWrm the role of PACAP in the pituitary and to elucidate the pleiotro-pic eVects of PACAP in Wsh. 
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Yuen-Yuen Kwok, Jessica Yan-Shuen Chu, Hubert Vaudry, Laurent Yon, Youssef Anouar, et al.. Cloning and characterization of a PAC1 receptor hop-1 splice variant in goldfish (Carassius auratus). General and Comparative Endocrinology, Elsevier, 2006, ⟨10.1016/j.ygcen.2005.08.011⟩. ⟨hal-02334629⟩



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