AVT and IT regulate ion transport across the opercular epithelium of killifish (Fundulus heteroclitus) and gilthead sea bream (Sparus aurata)

Abstract

The regulatory role of arginine vasotocin (AVT) and isotocin (IT) in Cl− secretion was investigated with the short circuit current (Isc) technique in opercular epithelia of killifish (Fundulus heteroclitus) and gilthead sea bream (Sparus aurata). Sea bream operculum showed ~ 4-fold lower number of Na/K-ATPase immunoreactive cells and ~ 12-fold lower secretory current than the killifish. In sea bream opercular membranes, the basolateral addition of AVT (10− 6 M) significantly stimulated Cl− secretion, while IT (10− 6 M) was without effect. In killifish, IT produced an immediate dose-dependent stimulation of Cl− secretion with significant effect at doses ≥ 10− 7 M and stimulation maxima (∆Isc ~ 25 μA ⋅ cm− 2) at 10− 6 M. The basolateral addition of bumetanide (200 μM) abolished > 75% of the effect of IT on Cl− secretion. In turn, AVT had a dual effect on killifish opercular Isc: an immediate response (~ 3 min) with Isc reduction in an inverted bell-shaped dose–response manner with higher current decrease (− 22 μA ⋅ cm− 2) at 10− 8 M AVT, and a sustained dose-dependent stimulation of Cl− secretion (stable up to 1 h), with a threshold significant effect at 10− 8 M and maximal stimulation (~ 20 μA ⋅ cm− 2) at 10− 6 M. Both effects of AVT appear receptor type specific. The V1-receptor antagonist SR 49059 abolished Isc reduction in response to AVT, while the specific V2-receptor antagonist (Tolvaptan, 1 μM) abolished the stimulatory action of AVT on Cl− secretion. According to these results, we propose a modulatory role for AVT and IT in Cl− (NaCl) secretion across the opercular epithelium of marine teleost.JAM-S was funded by Ministry of Education (Spain) through the program “Formación de Profesorado Universitario” (Ref: AP2008-01194). This work was partially supported by Ministry of Science and Education, Spain by Project AGL2010-14876 to JMM and by the Ministry of Science and Higher Education and European Social Funds through the Portuguese National Science Foundation by Projects PTDC/MAR/104008/2008 and PTDC/MAR-BIO/3811/2012 to JF.Peer reviewe