Increased gene expression of a facilitated diffusion urea transporter in the skin of the African lungfish (Protopterus annectens) during massively elevated post-terrestrialization urea excretion

Abstract

SUMMARYThe full-length cDNA sequence of a putative urea transporter(lfUT) of the facilitated diffusion UT-A type has been cloned from the African lungfish Protopterus annectens. The lfUT cDNA is 1990 bp in length and its open reading frame encodes a 409 amino acid long protein, with a calculated molecular mass of 44,723 Da. The sequence is closest to those of amphibians (∼65% amino acid homology), followed by mammals and elasmobranchs (∼60%), and then teleosts (∼50%). lfUT was clearly expressed in gill, kidney, liver, skeletal muscle and skin. Upon re-immersion in water after 33 days of air exposure(`terrestrialization'), lungfish exhibited a massive rise in urea-N excretion which peaked at 12–30 h with rates of 2000–5000 μmol-N kg–1 h–1 (versus normal aquatic rates of<130 μmol-N kg–1 h–1) and persisted until 70 h. This appears to occur mainly through the skin. Total `excess'urea-N excretion amounted to ∼81,000–91,000 μmol-N kg–1 over 3 days. By real-time PCR, there was no difference in lfUT expression in the ventral abdominal skin between aquatic ammoniotelic controls and terrestrialized lungfish immediately after return to water (0 h), and no elevation of urea-N excretion at this time. However, skin biopsies revealed a significant 2.55-fold elevation of lfUTexpression at 14 h, coincident with peak urea-N excretion. At 48 h, there was no longer any significant difference in lfUT mRNA levels from those at 0 and 14 h, or from aquatic fed controls. In accordance with earlier studies, which identified elevated urea-N excretion via the skin of P. dolloi with pharmacology typical of UT-A carriers, these results argue that transcriptional activation of a facilitated diffusion type urea transporter (lfUT) occurs in the skin during re-immersion. This serves to clear the body burden of urea-N accumulated during terrestrialization.