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[An antagonist to GnRH in the control of reproduction in teleost fish: dopaminergic inhibition. Ancestral origin and differential conservation within vertebrates?].

In mammals, the neurohormonal control of the pituitary gonadotropes is provided by the gonadoliberin GnRH. Several studies on teleost fish indicate that a single positive control by GnRH is not a general rule among vertebrates. Peter and colleagues presented the first evidence of an inhibitory neurohormonal factor, "GRIF" (gonadotropin-release inhibiting factor). They induced a preovulatory LH surge by injuring particular brain areas in the goldfish. Subsequent in vivo and in vitro studies identified dopamine as GRIF, and neuroanatomical investigations have demonstrated that dopaminergic neurones in the anterior preoptic area projecting to the pituitary represent the anatomical substrate for GRIF activity. An inhibitory role of dopamine on the control of LH and ovulation/spermiation has been evidenced in many adult teleosts, including its implications for aquaculture. However, dopamine does not play an inhibitory role in all adult teleosts. As regards the early stages of gametogenesis and especially the control of puberty, a role for dopamine has been suggested or rejected depending on species. The European eel has a unique life cycle with a long prepubertal stage, which has made it a useful model to demonstrate the key-role of dopamine in the control of puberty. Data from tetrapods suggest that the role of dopamine as a GRIF is not restricted to the teleosts, but that it may have an ancient evolutionary origin, and has been differentially conserved throughout vertebrate evolution.

Animals↗

Correlation between biochemical properties and adaptive diversity of skeletal muscle myofibrils and myosin of some air-breathing teleosts.

Functional properties of myofibrils and relative stability of myosin of five teleosts Channa punctata, Clarias batrachus, M astacembalus armatus, Labeo rohita and Catla catla adapted to different breathing modes were compared. Myofibrillar contractility and m-ATPase of air-breathing organ (ABO) possessing C.punctata and C. batrachus were low and least affected by pH in the range of 7.1-8.5. However, their myosin isoforms were relatively thermostable, more soluble at sub-neutral pH values, between 0.1 to 0.15 M KCl concentrations and less susceptible to a-chymotryptic digestion. In contrast, myofibrils and myosin of water-breather major carps L. rohita and C. catla were more contractile and susceptible to pH and salt concentrations. Thus, correlation between catalytic efficiency and relative stability of myofibrils and myosin of ABO-possessing teleosts was of reverse order and magnitude, as compared to water-breathers. Interestingly, myofibrils and myosin of the behavioral air-breather M. armnatus showed intermediate properties. The specific levels of m-ATPase of all the five teleosts were in conformity with the levels of metabolic marker, the lactate dehydrogenase. The effect of chymotryptic cleavage of 94 and 173 kDa domains on ATPase, individuality of peptide maps of MyHC isomers and perturbation of phenylalanine residues by urea implicated hydrophobic residues in stabilizing myosin structure in these fish. The present study suggests two apparent evolutionary modifications of myofibrils and myosin in ABO-possessing teleosts: (i), 'down-regulation' of ATPase that explains sluggishness of such species and, (ii), more stable molecular structure to support stress of air-breathing modes of life.

Actomyosin↗

Interphotoreceptor retinoid-binding protein gene structure in tetrapods and teleost fish.

PURPOSE: The interphotoreceptor retinoid-binding protein (IRBP) gene possesses an unusual structure, encoding multiple Repeats, each consisting of about 300 amino acids. Our goals were to gain insight into the function of IRBP, and to test the current model for the evolution of IRBP, in which Repeats were replicated from a simpler ancestral gene. METHODS: We employed a bioinformatics approach to analyze IRBP loci in recently completed or near-complete genome sequences of several vertebrates and nonvertebrate chordates. IRBP gene expression in zebrafish was evaluated by reverse transcriptase PCR (RT-PCR) and in situ mRNA hybridizations with gene-specific probes. RESULTS: Patterns of exons and introns in the IRBP genes of tetrapods were highly similar, as were predicted amino acid sequences and Repeat structures. IRBP gene structure in teleost fish was more variable, and we report a new gene structure for two species, the Japanese puffer fish (Takifugu rubripes) and the zebrafish (Danio rerio). These teleost genomes contain a two-gene IRBP locus arranged head-to-tail in which the first gene, Gene 1, is intronless and contains a single large exon encoding three complete Repeats. It is followed by a second gene, Gene 2, which corresponds to the previously reported gene consisting of two Repeats spread across four exons and three introns. Each of the two zebrafish genes is transcribed. Gene 2 is expressed in the photoreceptors and RPE, and Gene 1 is expressed in the inner nuclear layer and weakly in the ganglion cell layer. CONCLUSIONS: The tetrapod IRBP gene structure is highly conserved while the teleost fish gene structure was a surprise: It appears to be a two-gene locus with distinct Repeat organization in each open reading frame. This gene structure and gene expression data are consistent with possible neofunctionalization or sub-function partitioning of Gene 1 and Gene 2 in the zebrafish. We suggest that the two-gene locus in teleost fish arose as a consequence of either the known whole genome duplication or single gene tandem duplication.

Animals↗

Comparative anatomy of the topography of catecholamine containing neuron system in the brain stem from birds to teleosts.

The distribution of catecholamine (CA) containing neurons and nerve terminals in the brain stem from birds to teleosts were studied by means of histofluorescence technique of glyoxylic acid-formaldehyde combining method. The present study revealed that there exists marked differences in topography of CA neurons between higher vertebrates (birds and reptile) and lower vertebrates (amphibia and teleosts). Tegmental CA neurons in the birds and reptile are much more developed than those of amphibia and teleosts. Furthermore, huge collection of CA neurons (presumably dopamine) in the midbrain tegmentum could not be found in the amphibia and teleosts. On the other hand, distribution of CA nerve terminals in the brain stem is essentially similar throughout the vertebrate brains, though there exist modifications by secondary pattern distortions which correspond to various conditions of their lives. Some possible significance of CA neurons on development of central nervous system was discussed.

Animals↗

Localization and pattern of the mitotic activity in the central nervous system of developing oviparous and viviparous teleosts.

The subject of this work is an analysis of the mitotic activity in the developing spinal cord and rhombencephalon of the oviparous teleost Jordanella floridae and the viviparous teleost Gambusia affinis (Cyprinodontoidei). In Gambusia there is considerable and long lasting mitotic activity that decreases gradually and is still present 20 days after birth. In Jordanella the mitotic activity is concentrated in the early stages of development, it then decreases drastically and practically disappears even before hatching and reappears 20 days later. These differences are probably due to the variations in speed and pattern of development in the two teleosts. As regard the localization of mitoses, it emerged that migration towards the lumen may be prevented by physiological conditions related to normal development. It emerged moreover, that migration towards the lumen is not a generalised phenomenon in development of the central nervous system since it is related, in the spinal cord of teleosts at least, to the modality and speed of embryo development.

Animals↗

Sonic/vocal-acousticolateralis pathways in teleost fishes: a transneuronal biocytin study in mochokid catfish.

Mochokid catfish have two sound producing (sonic) organs--a pectoral spine stridulatory apparatus and a swimbladder whose vibration is established by nearby "drumming" muscles. Dextran-biotin or biocytin application to sonic nerves or muscles identified topographically separated motoneuron pools. Pectoral spine-related motoneurons are located within the ventral motor column whereas swimbladder motoneurons lie just ventral to the central canal or fourth ventricle. Axons of both groups of motoneurons exit the brain and spinal cord via ventral roots of occipital (swimbladder and pectoral) and spinal (swimbladder only) nerves. Transneuronal biocytin transport identified an extensive premotor network only for the swimbladder motor nuclei. Premotoneuron somata are located ipsilaterally in 1) a dorsolateral region of the sonic motor nucleus (SMN); motoneurons were clustered in the ventromedial region of the SMN and 2) the ventromedial medulla at the rostral pole of the SMN. Biocytin-filled fibers and less frequently premotoneuron somata were also found in the contralateral SMN. Biocytin-labeled fibers were continuous farther rostrally with 1) a commissural bundle that terminated bilaterally in the medial reticular formation near the caudal pole of the descending octaval nucleus and 2) a lateral brainstem bundle that terminated ipsilaterally in regions of the medulla and cerebellum considered to subserve acoustic and lateral line functions. Together with other data in distantly related teleost fishes, the results support the hypotheses that 1) central pathways linking sound-generating (sonic or vocal) and acoustic regions of the brain are traits common to both teleosts fishes and tetrapods that actively generate sounds, and 2) sonic/vocal pathways in teleosts have a conserved pattern of organization suggestive of common developmental origins.

Animals↗

Trimethylamine oxide counteracts effects of hydrostatic pressure on proteins of deep-sea teleosts.

In shallow marine teleost fishes, the osmolyte trimethylamine oxide (TMAO) is typically found at <70 mmol/kg wet weight. Recently we found deep-sea teleosts have up to 288 mmol/kg, increasing in the order shallow < bathyal < abyssal. We hypothesized that this protein stabilizer counteracts inhibition of proteins by hydrostatic pressure, and showed that, for lactate dehydrogenases (LDH), 250 mM TMAO fully offset an increase in NADH K(m) at physiological pressure, and partly reversed pressure-enhanced losses of activity at supranormal pressures. In this study, we examined other effects of pressure and TMAO on proteins of teleosts that live from 2000-5000 m (200-500 atmospheres [atm]). First, for LDH from a grenadier (Coryphaenoides leptolepis) at 500 atm for 8 hr, there was a significant 15% loss in activity (P < 0.05 relative to 1 atm control) that was reduced with 250 mM TMAO to an insignificant loss. Second, for pyruvate kinase from a morid cod (Antimora microlepis) at 200 atm, there was 73% increase in ADP K(m) without TMAO (P < 0.01 relative to K(m) at 1 atm) but only a 29% increase with 300 mM TMAO. Third, for G-actin from a grenadier (C. armatus) at 500 atm for 16 hr, there was a significant reduction of F-actin polymerization (P < 0.01 compared to polymerization at 1 atm) that was fully counteracted by 250 mM TMAO, but was unchanged in 250 mM glycine. These findings support the hypothesis. J. Exp. Zool. 289:172-176, 2001.

Actins↗

Structural evidence for two different testicular types in teleost fishes.

Testicular structure in Salmoniformes, Perciformes, Cypriniformes, and Atheriniformes has been examined and reinterpreted on the basis of two different tubular types, distinguished from each other by the intratubular distribution of spermatogonia. In the salmoniform, perciform, and cypriniform teleosts studied, spermatogonia are distributed along the entire length of the testicular tubules. However, in the atheriniform teleosts spermatogonia are restricted to the distal end of the tubule. Sperm development in teleosts is cystic, cysts being comprised of Sertoli-cell processes. In both testicular types described, Sertoli cells phagocytize spermatid residual bodies. Together with the germ cells, they comprise the only intratubular cell types within the teleostean testis. Boundary cells are located immediately outside of the tubule basement membrane. They do not form a complete layer over the tubule surface; therefore, interstitial Leydig cells and blood vessels may border directly upon the tubular basement membrane.

Animals↗

TrkA and TrkB neurotrophin receptor immunoreactivity in the teleost (Scorpaena porcus) endocrine pancreas.

Mammalian-like and specific neurotrophins, as well as their cognate tyrosine kinase Trk-like receptors have been identified in teleosts. They are mainly distributed in neuronal tissues, but evidence suggests that some non-neuronal tissues also express Trks. In this study we used immunohistochemistry to investigate the occurrence and cell distribution of Trks in the pancreas of teleosts (Scorpaena porcus). Immunoreactivity for TrkA and TrkB, but not for TrkC receptors, was found in the scorpionfish pancreas. TrkA-like positive cells were exclusively observed within pancreatic islets, among insulin- and glucagon-containing cells, but apparently did not co-localize with these hormones. TrkB immunostaining was found in islet cells, presumably colocalized with glucagon, as well as in some cells of the exocrine portion. These data provide a morphological basis for a role of the TrkA and TrkB ligands in the endocrine pancreas of teleosts.

Animals↗

Ultrastructural and cytochemical study of elastic fibers in the ventral aorta of a teleost, Anguilla japonica.

Previous studies have revealed that amorphous elastin and microfibrils are structural entities of mammalian elastic fibers. Elastin shows a wide phylogenetic distribution, but the presence of elastin-associated microfibrils has not been demonstrated in teleost aorta. Thus, we have ultrastructurally and cytochemically examined elastic fibers in the ventral aorta of eel, a teleost, by utilizing routine uranyl acetate and lead double staining, the tannic acid (pH 7.0)-uranyl acetate (TA-UA) method, elastase en bloc digestion, Thiéry's periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method, and the horseradish-peroxidase-labeled concanavalin A (Con A) method. In the ventral aorta of eel, a little ultrastructural difference between elastic fibers in the intima and media and those in the adventitia was noticed, but in either tunic each elastic fiber was basically composed of a "fibrillar core" and surrounding microfibrils. The fibrillar core was a collection of fibrils which showed a tendency to coalesce with each other, and these constituent fibrils were TA-UA positive and elastase-sensitive, representing their nature of elastin. By contrast, microfibrils associated with the fibrillar core were TA-UA negative and elastase-resistant, and their glycoproteinaceous nature was demonstrated by PA-TCH-SP and Con A methods. Thus, this study provides evidence for the presence of elastin-associated microfibrils in teleost aorta. These results are discussed in relation to the topographical difference of elastic fibers in eel aortic wall.

Anguilla↗

Distribution and quantification of corticotropin-releasing hormone (CRH) in the brain of the teleost fish Oreochromis mossambicus (tilapia).

The recent characterization of the corticotropin-releasing hormone (CRH) prehormone of the fish tilapia (Oreochromis mossambicus) showed that more variation exists between vertebrate CRH amino acid sequences than recognized before. The present study investigates whether the deviating composition of tilapia CRH coincides with an atypical distribution of CRH in the brain. For this purpose we applied immunohistochemistry, as well as radioimmunoassay (RIA) quantification in brain slices. The results are plotted in a new atlas and reconstruction of the tilapia brain. The largest population of CRH-immunoreactive (ir) neurons is present in the lateral part of the ventral telencephalon (Vl). Approximately tenfold less CRH-ir neurons are observed in the preoptic and tuberal region. The CRH-ir neurons observed in the preoptic region are parvocellular and do not, or hardly, display arginine-vasotocin (AVT) immunoreactivity. CRH-ir neurons are also present in the glomerular layer of the olfactory bulb, in the periventricular layer of the optic tectum, and caudal to the glomerular nucleus. A very dense plexus of CRH-ir terminals is located in the most rostral part of the dorsal telencephalon. This region has not been described in other teleosts and is in the present study subdivided into the anterior part of the dorsal telencephalon (Da) and the anterior part of the laterodorsal telencephalon (Dla). High densities of CRH-ir terminals were observed in and around Vl, in the tuberal region, around the rostral part of the lateral recess, and in the caudal part of the vagal lobe. In the pituitary, CRH-ir terminals are concentrated in the neuro-intermediate lobe. Overall, the immunohistochemical and quantitative data correlated well, as the RIA CRH profile in serial 160-microm slices revealed four peaks, which corresponded with major ir-cell groups and terminal fields. Our results strongly suggest that the CRH-ir cells of Vl project to the rostro-dorsal telencephalon. Consequently, they may not be primarily involved in regulation of pituitary cell types but may subserve other functions. The presence of a CRH-containing Vl-Da/Dla projection seems to be restricted to the most modern group of teleosts, i.e., the Acanthopterygians. Further anatomic indications for non-pituitary-related functions of CRH are found in the vagal lobe and the optic tectum of tilapia. Although the low CRH content of the preoptic region reported here for tilapia may be typical for unstressed fish, the fact remains that remarkably few CRH-ir neurons are involved in regulating the pituitary. Overall, the CRH distribution in the brain of tilapia is more widespread than previously reported for other teleosts.

Animals↗

Isolation and characterization of two teleost melanopsin genes and their differential expression within the inner retina and brain.

Melanopsin is a newly discovered photopigment that is believed to be involved in the regulation of circadian rhythms in tetrapods. Here we describe the characterization of the first two teleost melanopsins (opn4a and opn4b) isolated from Atlantic cod (Gadus morhua). These two teleost genes belong to a subgroup of melanopsins that also include members from Xenopus, chicken, and Takifugu. In situ hybridization revealed that opn4a and opn4b are differentially expressed within the retina and brain. In the larval and adult retina, both melanopsins are expressed in a subset of cells in the inner retina, resembling amacrine and ganglion cells. In addition, opn4a is expressed in the horizontal cells, indicating a separate task for this gene. In the brain, the two melanopsins are separately expressed in two major retinal and extraretinal photosensitive integration centers, namely, the suprachiasmatic nucleus (opn4a) and the habenula (opn4b). The expression of opn4a in the suprachiasmatic nucleus in cod is similar to the melanopsin expression found in Xenopus. This suggests a conserved role for this opsin and an involvement in mediation of nonvisual photoreceptive tasks, such as entraining circadian rhythms and/or hypophysiotrophic systems. The differential expression of opn4b in the habenula suggests that this gene plays a role similar to that of opn4a, in that it is also situated in an area that integrates photic inputs from the pineal as well as other brain regions. Thus, the habenula may be an additional region that mediates photic cues in teleosts.

Amino Acid Sequence↗

Form and function in the unique inner ear of a teleost: the silver perch (Bairdiella chrysoura).

Members of the teleost family Sciaenidae show significant variation in inner ear and swim bladder morphology as well as in the relationship between the swim bladder and the inner ear. In the silver perch (Bairdiella chrysoura), a Stellifer-group sciaenid, both the saccular and utricular otoliths are enlarged relative to those in other teleosts. Additionally, its swim bladder is two-chambered, and the anterior chamber surrounds the otic capsule and terminates lateral to the saccules. Structure and function of the auditory system of the silver perch were explored by using gross dissections, scanning electron microscopy, CT scan reconstruction, and auditory brainstem response approach. Several morphological specializations of the auditory system of the silver perch were found, including expansion of the utricular and lagenar otoliths, close proximity between the saccules and the utricles, deeply grooved sulci on the saccular otoliths, two-planar saccular sensory epithelia, and a unique orientation pattern of sensory hair cell ciliary bundles on the saccular sensory epithelium. It was determined that the silver perch can detect up to 4 kHz, with lowest auditory thresholds between 600 Hz and 1 kHz. Audition in the silver perch is comparable to that in the goldfish (Carassius auratus), a hearing "specialist." The morphological specializations of the inner ear and swim bladder of the silver perch may be linked to its enhanced hearing capabilities. The findings of this study support the proposal that sciaenids are excellent model species for investigating structure-function relations in the teleost auditory system.

Air Sacs↗

The basic helix-loop-helix transcription factor neuroD is expressed in the rod lineage of the teleost retina.

Persistent rod genesis in the retinas of teleost fish was first described over 2 decades ago, but little is known regarding the underlying genetic and molecular mechanisms that govern this phenomenon. Because of its function in the developing mammalian retina and persistently mitotic adult tissues, we sought to characterize the cellular expression of the basic helix-loop-helix (bHLH) transcription factor neuroD in the persistently neurogenic retina of adult teleosts. We show here that, in the adult retina of the goldfish, neuroD is expressed by putative amacrine cells, nascent cones, and the mitotically active cells of the rod lineage. neuroD is the first gene shown to be expressed by rod precursors, the immediate antecedents of rod photoreceptors. In contrast to the vertebrate classes described previously, neuroD is not expressed in multipotent progenitors in the teleost retina. Combining neuroD in situ hybridizations with cell-cycle-specific markers suggests that, in rod precursors, neuroD expression is cell cycle specific.

Animals↗

Distribution of serotonin in the brain of the mormyrid teleost Gnathonemus petersii.

The distribution of serotonin-immunoreactive neurons and fibers was studied in the highly developed brain of the weakly electric fish Gnathonemus petersii with the aid of specific antibodies against serotonin. Serotoninergic cell bodies occur in three regions: the raphe region of the brainstem, the hypothalamus, and the transition zone between the dorsal thalamus and the pretectum. Serotoninergic raphe neurons are clustered in three groups: nucleus raphes superior, intermedius, and inferior. The latter has not been described in other teleosts and thus might be the source of the serotoninergic innervation of specific mormyrid electrosensory brain regions. Most hypothalamic serotoninergic neurons have cerebrospinal-fluid (CSF)-contacting processes and thus belong to the paraventricular organ (PVO), which in Gnathonemus is located around a number of small infundibular recesses. The distribution of serotonin in the PVO precisely matches the distribution of dopamine, as described previously. Serotoninergic cells in the thalamopretectal transition zone also have been described in other teleosts, but not in other vertebrate groups, and thus seem to represent a teleostean specialization. Serotoninergic fiber density is especially high in the medial forebrain bundle and surrounding preoptic and hypothalamic regions as well as in several telencephalic and preoptic subependymal plexus. Serotoninergic fibers appear to be almost completely absent in the large and differentiated corpus and valvula cerebelli. Comparison with the literature on teleostean serotoninergic innervation patterns reveals several mormyrid specializations, including the absence of serotonin in large parts of the mormyrid telencephalic lobes, a differentiated innervation pattern of distinct electrosensory and mechanosensory subnuclei of the torus semicircularis, a refined serotoninergic lamination pattern in the midbrain tectum, and a prominent innervation of the electrosensory lateral line lobe, the associated caudal cerebellar lobe, and the electromotor medullary relay nucleus. A distinct innervation of several types of (pre)motor neurons, such as the Mauthner cells and facial motor neurons, has not been reported previously for other teleosts. Consequently, the distribution of serotoninergic fibers as well as neurons in the mormyrid brain is substantially adapted to the high degree of differentiation of its electrosensory and telencephalic brain regions, but serotoninergic innervation is not involved in the circuitry of the most impressive part of the mormyrid brain; i.e., its large corpus and valvula cerebelli.

Animals↗

Identification of cyclic nucleotide-regulated phosphoproteins, including phosducin, in motile rod inner-outer segments of teleosts.

In teleost retinas, rods elongate in the light and shorten in the dark. Rod motility is mediated by the actin cytoskeleton of the inner segment and is regulated by cyclic AMP- or cyclic GMP-stimulated phosphorylation of target proteins. In this study, we have identified the target proteins of cyclic nucleotide-dependent kinases in rods, using preparations of isolated, motile rod inner-outer segments (RIS-ROS). Five proteins found in Percoll-purified RIS-ROS were phosphorylated in the presence of cAMP (> 10 nM), cGMP (> or = 10 microM) and exogenous catalytic subunit of cAMP-dependent protein kinase (PKA). The PKA inhibitor, PKI, blocked stimulation of phosphorylation by both cAMP and cGMP. Three cAMP-stimulated phosphoproteins were detected in cytoskeletal fractions of light- and dark-adapted RIS-ROS. One of these, PP33, appears to be a fish homologue of mammalian phosducin, based on immunolabeling by two different antibodies against mammalian phosducin and on electrophoretic characteristics in 2-D gels. Two additional phosducin immunoreactive bands were detected in Western blots. One, at 35 kDa, comigrated with a second cAMP-stimulated RIS-ROS phosphoprotein, PP35, which was also detected in the cytoskeleton. The other, at 37 kDa, was present in whole teleost retinas but not in purified RIS-ROS. Our results suggest that the effects of both cAMP and cGMP on teleost rod motility are mediated through PKA modulation of target phosphoproteins. These phosphoproteins include a cytoskeleton-associated phosducin homologue.

Animals↗

GLUT-4 Deficiency and severe peripheral resistance to insulin in the teleost fish tilapia.

Teleost fish, in general, are glucose intolerant; this trait has been attributed to piscine islets secreting insulin primary in response to amino acid secretogogues rather than glucose. However, pancreatic islet from the teleost fish tilapia, when transplanted into diabetic nude mice, were glucose responsive even though tilapia were severely glucose intolerant. This suggested a strong peripheral resistance to the glucostatic effects of insulin. Using Western blotting with polyclonal antibodies as well as Northern analysis for mRNA, tilapia tissues were found to be devoid of GLUT-4, the insulin-sensitive glucose transporter responsible for the hypoglycemic effect of insulin in mammals. The absence of GLUT-4 in peripheral tissues may explain why tilapia, and possibly other teleost fish, are severely glucose intolerant. This suggests that tilapia islets have evolved along mammalian lines to be glucose sensitive while tilapia peripheral tissue have diverged widely. Using the same methods, tilapia were found to have a very limited tissue distribution of the insulin-independent glucose transporter, GLUT-1, which is responsible for basal glucose transport in mammalian cells. It is suggested that tilapia provide a naturally occurring GLUT-4 knockout model.

Adipose Tissue↗

Characterization of teleost insulin receptor family members. II. Developmental expression of insulin-like growth factor type I receptor messenger RNAs in rainbow trout.

The insulin-like growth factor (IGF) system in teleosts consists of two ligands, IGF-I and IGF-II, multiple binding proteins, and high-affinity transmembrane receptors. There exists a large gap in our knowledge of the structure and expression of receptors mediating the biological effects of the IGFs in teleosts. For example, nucleotide sequence data other than those from the kinase domain, evidence of multiple genes, mRNA expression pattern and polyadenylation status in multiple tissues at different developmental stages, and quantitation of mRNA levels in multiple tissues are not known for any teleost. In the study described here, two rainbow trout IGF type I receptor cDNAs (rtIGFR Ia and rtIGFR Ib) were isolated by a 5' rapid amplification of cDNA ends method and confirmed as separate genes by genomic Southern blot hybridization. The predicted amino acid sequences are 85% identical to each other in the tyrosine kinase domain. Both cDNAs are more homologous to mammalian IGF type I receptors than to insulin receptors. Reverse transcription-polymerase chain reaction from total RNA using either oligo(dT) or random hexamers as primers resulted in a diminished ability to detect IGF receptor mRNAs when oligo(dT) was used, suggesting developmental and tissue-specific polyadenylation. The highest steady-state mRNA levels of rtIGFR Ia were found in juvenile gill and adult heart, while the highest levels of rtIGFR Ib were found in adult pyloric caeca, which also contained diffuse pancreatic and adipose tissue. The lowest steady-state mRNA levels of both rtIGFR Ia and rtIGFR Ib were found in juvenile heart, liver, muscle, and spleen, and adult liver. Significant differences in steady-state mRNA levels were also found between juveniles and adults. These results suggest a complex expression pattern of IGF type I receptor mRNAs in partial tetraploid fish.

Amino Acid Sequence↗