Whole-cell response characteristics of ciliated and microvillous olfactory receptor neurons to amino acids, pheromone candidates and urine in rainbow trout.

Abstract

Olfactory lamellae of teleosts contain two morphologically different types of olfactory receptor neurons (ORNs): ciliated ORNs (cORNs) and microvillous ORNs (mORNs). However, little is known about the functional difference between these two types of ORNs in fish olfaction. We isolated cORNs and mORNs using a Ca(2+)-free solution method from olfactory organs of the rainbow trout and examined their response characteristics to various odorants including fish pheromone candidates by whole-cell voltage-clamp techniques. Quadruple mixture of amino acids, single amino acids, steroids (analogues of DHP; 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one and ECG; etiocholan-3 alpha-ol-17-one glucuronide), prostaglandins (PGFs) and urine samples collected from immature and mature female fish were applied focally to olfactory cilia or microvilli using a multi-barreled stimulation pipette with a pressure ejection system. Inward current responses to odorants were recorded from both cORNs and mORNs at a holding potential of -60 mV. cORNs responded to the amino acid mixture, single amino acids, urine samples and ECG, whereas mORNs responded specifically either to the amino acid mixture or single amino acids. The response profiles of both cORNs and mORNs to various odorants varied widely. None of cORNs and mORNs responded to fish pheromone candidates, PGFs and DHPs. Androgen treatment of immature fish did not influence olfactory sensitivity of both cORNs and mORNs to the amino acid mixture and both urine samples. Amino acid and bile acid analyses by HPLC showed that both urine samples contained 35 amino acids (1-40 mM) and trace amounts of taurocholic acid and glycoursodeoxycholic acid. Our results suggest that cORNs are 'generalists' that respond to a wide variety of odorants, including pheromones, whereas mORNs are 'specialists', specific to amino acids, and also suggest that PGFs and DHPs are not pheromones for the rainbow trout.