The olfactory epithelium of fish contains three intermingled types of olfactory receptor neurons (ORNs): ciliated, microvillous, and crypt. The present experiments were undertaken to test whether the different types of ORNs respond to different classes of odorants via different families of receptor molecules and G-proteins corresponding to the morphology of the ORN. In catfish, ciliated ORNs express OR-type receptors and Galpha(olf). Microvillous ORNs are heterogeneous, with many expressing Galpha(q)/11, whereas crypt ORNs express Galpha(o). Retrograde tracing experiments show that ciliated ORNs project predominantly to regions of the olfactory bulb (OB) that respond to bile salts (medial) and amino acids (ventral) (Nikonov and Caprio, 2001). In contrast, microvillous ORNs project almost entirely to the dorsal surface of the OB, where responses to nucleotides (posterior OB) and amino acids (anterior OB) predominate. These anatomical findings are consistent with our pharmacological results showing that forskolin (which interferes with Galpha(olf)/cAMP signaling) blocks responses to bile salts and markedly reduces responses to amino acids. Conversely, U-73122 and U-73343 (which interfere with Galpha(q)/11/phospholipase C signaling) diminish amino acid responses but leave bile salt and nucleotide responses essentially unchanged. In summary, our results indicate that bile salt odorants are detected predominantly by ciliated ORNs relying on the Galpha(olf)/cAMP transduction cascade. Nucleotides are detected by microvillous ORNs using neither Galpha(olf)/cAMP nor Galpha(q)/11/PLC cascades. Finally, amino acid odorants activate both ciliated and microvillous ORNs but via different transduction pathways in the two types of cells.

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Electrophysiological experiments indicate that olfactory receptors of the channel catfish, Ictalurus punctatus, contain different receptor sites for the acidic (A), basic (B), and neutral amino acids ; further, at least two partially interacting neutral sites exist, one for the hydrophilic neutral amino acids containing short side chains (SCN), and the second for the hydrophobic amino acids containing long side chains (LCN) . The extent of cross-adaptation was determined by comparing the electro-olfactogram (EOG) responses to 20 "test" amino acids during continuous bathing of the olfactory mucosa with water only (control) to those during each of the eight "adapting" amino acid regimes. Both the adapting and test amino acids were adjusted in concentrations to provide approximately equal response magnitudes in the unadapted state . Under all eight adapting regimes, the test FOG responses were reduced from those obtained in the unadapted state, but substantial quantitative differences resulted, depending upon the molecular structure of the adapting stimulus . Analyses of the patterns of EOG responses to the test stimuli identified and characterized the respective "transduction processes," a term used to describe membrane events initiated by a particular subset of amino acid stimuli that are intricately linked to the origin of the olfactory receptor potential . Only when the stimulus compounds interact with different transduction processes are the stimuli assumed to bind to different membrane "sites." Four relatively independent L-a-amino acid transduction processes (and thus at least four binding sites) identified in this report include : (a) the A process for aspartic and glutamic acids ; (b) the B process for arginine and lysine ; (c) the SCN process for glycine, alanine, serine, glutamine, and possibly cysteine ; (d) the LCN process for methionine, ethionine, valine, norvaline, leucine, norleucine, glutamic acid-7-methyl ester, histidine, phenylalanine, and also possibly cysteine . The specificities of these olfactory transduction processes in the catfish are similar to those for the biochemically determined receptor sites for amino acids in other species of fishes and to amino acid transport specificities in tissues of a variety of organisms .Address reprint requests to Dr.

Horseradish peroxidase tracing and extracellular electrophysiological recording techniques were employed to delineate prosencephalic connections of the gustatory system in ictalurid catfishes. The isthmic secondary gustatory nucleus projects rostrally to several areas of the ventral diencephalon including the nucleus lobobulbaris and the nucleus lateralis thalami. Injections of HRP in the vicinity of the nucleus lobobulbaris reveal an ascending projection to the telencephalon terminating in the area dorsalis pars medialis (Dm) and the medial region of area dorsalis pars centralis (Dc). Conversely, injections of HRP into the gustatory region of area dorsalis pars medialis label small neurons in the nucleus lobobulbaris. Gustatory neurons in the telencephalon send descending projections via the medial and lateral forebrain bundles to several nuclei in the anterior and ventroposterior diencephalon. The nucleus lateralis thalami, a diencephalic nucleus, receives ascending gustatory projections from the secondary gustatory nucleus but does not project to the telencephalon. Neurons in both the nucleus lateralis thalami and the telencephalic gustatory target exhibit multiple extraoral and oral receptive fields and complex responses to chemical (taste) and tactile stimulation.

SUMMARY Electrophysiological responses of goldfish olfactory receptor neurons(ORNs) and goldfish behavioral responses to polyamines were investigated in vivo. Electro-olfactogram (EOG) recordings indicated that polyamines (putrescine, cadaverine and spermine) are potent olfactory stimuli for goldfish with estimated electrophysiological thresholds of 10–8–10–7 mol l–1,similar to that for L-arginine, the most stimulatory amino acid. Although thresholds were similar, the magnitude of the EOG responses to intermediate(10–5–10–4 mol l–1)and high (10–3 mol l–1) concentrations of polyamines dwarfed the responses to amino acids and related single amine containing compounds (amylamine and butylamine). The EOG responses to 0.1 mmol l–1 putrescine, cadaverine and spermine were, respectively,4.2×, 4.3× and 10.3× the response of the standard, 0.1 mmol l–1 L-arginine. Electrophysiological cross-adaptation experiments indicated that polyamine receptor sites are independent from those to L-amino acids (alanine, arginine, glutamate, lysine, methionine and ornithine), bile salts (sodium taurocholate and taurolithocholate), the single amine containing compounds (amylamine and butylamine) and ATP. Further, the cross-adaptation experiments revealed the existence of independent receptor sites for the different polyamines tested. Pharmacological experiments suggested that polyamine odorant transduction does not primarily involve the cyclic AMP and IP3 second messenger pathways. Behavioral assays indicated that polyamines are attractants that elicit feeding behavior similar to that elicited by L-amino acids.

Vertebrate gustatory systems include a tertiary ascending pathway from a secondary gustatory nucleus in the hindbrain to several forebrain nuclei. This connection is prominent in catfish, corresponding to their highly developed sense of taste. Iontophoretic injections of horseradish peroxidase were used to identify the specific target nuclei of the tertiary gustatory pathway in channel catfish and to characterize those nuclei by their respective connections. Efferents from the secondary gustatory nucleus (nGS) ascend in the tertiary gustatory tract to the caudal inferior lobe, where they terminate caudally in the nucleus lobobulbaris (nLB) and nucleus centralis (nCLI), and rostrally in the nucleus diffusus (nDLI). Secondary projections from the facial lobe (FL) also terminate in the nLB and in the nucleus subglomerulus (nSG). The nLB forms three cell groups (caudal--nLB, rostrolateral--rl nLB, parvicellular--nLBp), which project to the facial lobe, vagal lobe, and telencephalon, respectively. Cells from the nCLI project throughout the caudal inferior lobe and to the acousticolateral torus semicircularis and telencephalon, while the nDLI and nSG have intrinsic connections within the inferior lobe. The lateral thalamic nucleus projects from this region back to the nGS. Through these identified connections several mechanisms for the processing of gustatory information can be proposed. The descending projections from the nLB and nLT could provide feedback to the primary and secondary gustatory nuclei, and could modulate feeding-related motor circuits in the medulla. The connections of nCLI and nLBp with the telencephalon allow for the involvement of gustation in learning processes and other complex behaviors.

1. We report for the first time in any teleost, a quantitative in vivo study of recordings from single olfactory receptor neurons (ORNs) in the channel catfish, Ictalurus punctatus, with odorant stimuli. 2. Responses of 69 spontaneously active single ORNs were recorded simultaneously with the electroolfactogram (EOG). Recording times ranged from 10 to 72 min per receptor cell with an average of 24 +/- 15 (SD) min/cell. The averaged spontaneous frequency ranged from < 1 to 12 action potentials/s with a mean frequency of 4.7 +/- 2.5 action potentials/s. 3. Catfish ORNs responded to the odorant stimuli (amino acids, bile salts, and ATP) with either an excitation or suppression of the background neural activity. Suppressive responses were encountered more frequently than excitatory responses, suggesting that suppressive responses also play an important role in olfactory coding. 4. Excitatory and suppressive responses to the different odorants were elicited from the same ORN, suggesting that different olfactory receptor molecules and different transduction pathways exist in the same ORN.

Extracellular electrophysiological recordings from single olfactory bulb (OB) neurons in the channel catfish, Ictalurus punctatus, indicated that the OB is divided into different functional zones, each processing a specific class of biologically relevant odor. Different OB regions responded preferentially at slightly above threshold to either a mixture of 1) bile salts (10(-7) to 10(-5) M Na(+) salts of taurocholic, lithocholic, and taurolithocholic acids), 2) nucleotides [10(-6) to 10(-4) M adenosine-5'-triphosphate (ATP), inosine-5'-monophosphate (IMP), and inosine-5'-triphosphate (ITP)], or 3) amino acids (10(-6) to 10(-4)M L-alanine, L-methionine, L-arginine, and L-glutamate). Excitatory responses to bile salts were observed primarily in a thin, medial strip in both the dorsal (100-450 microm) and ventral (900-1,200 microm) OB. Excitatory responses to nucleotides were obtained primarily from dorsal, caudolateral OB, whereas excitatory responses to amino acids occurred more rostrally in the dorsolateral OB, but continued more medially in the ventral OB. The chemotopy within the channel catfish OB is more comparable to that previously described by optical imaging studies in zebrafish than by field potential studies in salmonids. The present results are consistent with recent studies, suggesting that the specific spatial organization of output neurons in the OB is necessary for the quality coding/decoding of olfactory information.