Immunohistochemical analysis of the distribution of molecules involved in ionic and pH regulation in the lancelet Branchiostoma floridae (Hubbs, 1922)

Abstract

The aim of present work is to analyse the distribution of carbonic anhydrase II (CAII), cystic fibrosis transmembrane regulator (CFTR), vacuolar-type H⁺-ATPase (V-H⁺-ATPase), Na⁺/K⁺ ATPase, Na⁺/H⁺ exchanger (NHE) and SLC26A6 (solute carrier family 26, member 6), also known as pendrin protein, in the lancelet Branchiostoma floridae in order to go in depth in the evolution of osmoregulation and pH regulation in Chordates. In view of their phylogenetic position, lancelets may indeed provide a critical point of reference for studies on osmoregulation evolution in Chordates. The results of present work demonstrated that, except to Na⁺/K⁺ ATPase that is strongly expressed in nephridia only, all the other studied molecules are abundantly present in skin, coelomic epithelium, renal papillae and nephridia and hepatic coecum. Thus, it is possible to hypothesize that also in lancelet, as in fish, these organs are involved in pH control and ionic regulation. In the digestive tract of B. floridae, the intestine epithelium was weakly immune-reactive to all tested antibodies, while the hepatic coecum showed an intense immunoreactivity to all molecules. Since in amphioxus the hepatic coecum functions simultaneously as stomach, liver and pancreas, these immunohistochemical results proved the secretion of H+ and HCO₃^- ions, typical of digestive process. Colocalization studies indicated a co-expression of the studied proteins in all considered organs, excluding NHE and pendrin for renal papillae, since some renal papillae are NHE immunopositive only.

Keywords: Carbonic anhydrase II; Cystic fibrosis transmembrane regulator; Na(+)/H(+) exchanger; Na(+)/K(+) ATPase; Pendrin; Vacuolar-type H(+)-ATPase.