Expression of H⁺,K⁺-ATPase and Glycopattern Analysis in the Gastric Glands of Rana esculenta

Maria Mastrodonato ¹, Giuseppe Calamita ¹, Roberta Rossi ¹, Giovanni Scillitani ¹, Giuseppa Esterina Liquori ¹ and Domenico Ferri ^1*

¹ Dipartimento di Zoologia, Laboratorio di Istologia e Anatomia comparata (MM,GS,GEL,DF), Dipartimento di Fisiologia Generale ed Ambientale (GC), and Dipartimento di Anatomia Patologica, Servizio di Patologia Ultrastrutturale, Policlinico (RR), Università degli Studi di Bari, Bari, Italy

^* To whom correspondence should be addressed. E-mail: d.ferri{at}biologia.uniba.it.

Submitted on July 3, 2008
Accepted on 20 October 2008

	Abstract

A multidisciplinary study involving lectin histochemistry, immunohistochemistry, immuno-lectin blotting and immunogold was carried out to determine the distribution of sugar residues in the glycoproteins of Rana esculenta oxynticopeptic cells. We considered animals in two experimental conditions, fasting and fed. It is known that in mammals the tubulovesicular membranes are rich in proteins with several functions. The proton pump H⁺,K⁺-ATPase, a heterodimeric complex with a catalytic -subunit and a heavily glycosylated -subunit, responsible for acid secretion, is the most abundant. No data have been published regarding the localization and the structures of H⁺,K⁺-ATPase in amphibians. In the water frog, the luminal membrane and tubulovesicular system of oxynticopeptic cells, which differ in morphology according to their functional stage, reacted with the primary gold-conjugated antibody against the H⁺,K⁺-ATPase -subunit. By lectin histochemistry and immunoblotting, in the oxynticopeptic cells of R. esculenta we detected the presence of N-linked glycans having fucosylated (poly)lactosamine chains, which could correspond to the oligosaccharide chains of the subunit. The latter are somewhat different from those described in mammals, and this is probably due to an adaptation to the different microenvironmental conditions in which the oxynticopeptic cells find themselves, in terms of their different habits and phylogeny.

Key Words: glycohistochemistry, lectins, oxynticopeptic cells, digestive tract, Rana esculenta

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