Current knowledge of ghrelin in amphibians

We are exploring physiological importance of the ghrelin system in vertebrates. This review summarizes current knowledge of the ghrelin system in amphibians. Our study on ghrelin precursor in various amphibians revealed that the third amino acid with acyl modification has changed to threonine (Thr-3) instead of serine (Ser-3) only in the genus, Rana. Functional analyses of the ghrelin receptor in three species of amphibians, Japanese fire belly newt, American bullfrog and Japanese tree frog revealed that ghrelin and GHS-R1a agonists increase intracellular Ca²⁺ concentration in HEK293 cells expressing each receptor, and that ligand selectivity of ghrelin with Ser-3 and Thr-3 that expected to see in the bullfrog receptor was not found in the two frog receptors, but in the newt receptor. The brain, gastrointestinal tract, kidney and gonad highly express GHS-R1a mRNA. In frogs and newt, fasting did not increase GHS-R1a mRNA expression in the brain, but in the stomach. However, intraperitoneal (IP) injection of ghrelin did not affect food intake. A dehydration treatment increased GHS-R1a mRNA expression in the brain, stomach and ventral skin in the tree frog. However, intracerebroventricular (ICV) injection of ghrelin did not affect water absorption. Ghrelin did not influence gastrointestinal motility in in vitro studies using smooth muscle strips of the bullfrog and newt in vitro. These results suggest that the ghrelin system is present in various amphibians, but little is known about the physiological functions except hypophyseal hormone secretion.