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PubMed1995

Teleost isotocin receptor: structure, functional expression, mRNA distribution and phylogeny.

A cDNA encoding a receptor for the oxytocin-related peptide isotocin has been identified by screening a lambda gt11 library constructed from poly(A)+ RNA of the hypothalamic region of the teleost Catostomus commersoni. The probe used was obtained by PCR amplification of white sucker genomic DNA using degenerate primers based on conserved sequences in the mammalian receptor counterparts. The full-length cDNA specifies a polypeptide of 390 amino acid residues that displays the typical hydrophobicity profile of a seven transmembrane domain receptor and which exhibits greatest similarity to mammalian oxytocin receptors. Oocytes that express the cloned receptor respond to the application of isotocin by an induction of membrane chloride currents indicating that it is coupled to the inositol phosphate/calcium pathway. The isotocin receptor (ITR) can also be activated by vasotocin, mesotocin, oxytocin and Arg-vasopressin, although these have lower potencies than isotocin. ITR-encoding mRNA has been detected in brain, intestine, bladder, skeletal muscle, lateral line, gills and kidney indicating that this receptor may mediate a variety of physiological functions.

Amino Acid Sequence↗
PubMed1983

[Ultrastructure of prolactin cells and survival of Gambusia sp. (teleost fish) in deionized water enriched in calcium or sodium].

The ultrastructure of the prolactin (PRL) cells of Gambusia was studied in animals kept in deionized water (EDes) and in EDes supplemented either with Ca2+ (10,2 and 15,3 mM) or with Na+ (10,2 and 15,3 mM). In environments supplemented with CaCl2 the maximal survival was 40 days. The stimulation of PRL cells was similar to that described in EDes. On the contrary, in environments supplemented with NaCl, 50% of the animals are still alive after 45 days. PRL cells are slightly stimulated and their ultrastructural aspect is similar to that observed in controls kept in freshwater. The significance of these results is discussed in relation with our present knowledge of the pituitary control of osmoregulation in teleost fish.

Animals↗
PubMed1984

The vascular system of principal islets: semithin-section studies in teleosts fixed by perfusion.

The angioarchitecture of clearly zonated principal islets (central B-cell and peripheral A-cell region) from teleosts has been analyzed in five islets following perfusion fixation and serial sectioning. In contrast to the similar cell-type distribution in the species studied (Gadus morrhua , Myoxocephalus scorpius , Agonus cataphractus , Limanda limanda ) there were three different types of vasculature characterized by centripetal blood flow, centrifugal blood flow, or an intermediate type. The vascular pattern (and the development of the organ capsule) showed inter- and even intraspecies differences. It is suggested that differences in angioarchitecture reflect individually variable interactions of exocrine and endocrine tissue during organogenesis rather than mutual cell-type control via the vascular system.

Animals↗
PubMed1985

Somatostatin-like immunoreactivity in the pituitary and brain of three teleost fish species: somatostatin as a potential regulator of prolactin cell function.

Somatostatin-like immunofluorescence occurs in the hypothalamus and neurohypophysis of three euryhaline teleosts: tilapia, killifish, and mudsucker. This immunofluorescence was eliminated by incubating the primary antibody with excess somatostatin or somatostatin-28 but not with urotensin II, a partial analogue of somatostatin. In all three fishes, the strongest reaction was seen in the proximal pars distalis and parts of the pars intermedia. Strongly fluorescing processes from cells of the preoptic nucleus extend toward the pituitary. Distinct fluorescence was also associated with the neurohypophysis penetrating into the rostral pars distalis in the tilapia but not in the killifish or mudsucker. In the tilapia, an extensive network of immunofluorescent fibers and small cells were present in the anterior dorsolateral telencephalon, in addition to a moderately fluorescing group of cells anterolateral to the preoptic nucleus. A small area of diffuse fluorescence was also seen in the anterior dorsolateral midbrain tegmentum. Previous physiological studies have implicated somatostatin as a regulator of prolactin cell activity in tilapia. The present study demonstrates the route by which somatostatin may be delivered to the rostral pars distalis to inhibit prolactin secretion.

Animals↗
PubMed1986

Action of melanin-concentrating hormone (MCH) on teleost chromatophores.

The in vitro effects of synthetic salmon melanin-concentrating hormone (MCH) on chromatophores of four teleost species were studied. In the erythrophores of the platyfish (Xiphophorus maculatus) and the swordtail (Xiphophorus helleri), and in the xanthophores and amelanotic melanophores of the medaka (Oryzias latipes), pigment aggregation took place in response to MCH even in the absence of Ca2+. In contrast to this, the leucophores of the medaka responded to MCH by the pigment dispersion but only when Ca2+ was present. The motile iridophores of the blue damselfish (Chrysiptera cyanea), which play a predominant role in coloration and its changes, were not affected by the hormone. Pharmacological studies employing various blocking agents suggest that the pigment-aggregating action of MCH is probably mediated through specific receptors possessed by the erythrophores, xanthophores, or amelanotic melanophores, while the pigment-dispersing action on the leucophores might be revealed through the receptors for melanophore-stimulating hormone (MSH).

Animals↗
PubMed1986

Sodium-retaining activity of chum salmon prolactin in some euryhaline teleosts.

Sodium-retaining activity of chum salmon prolactin (PRL) was examined in several euryhaline teleosts. Chum PRL was 100 times more potent than ovine PRL in maintaining plasma sodium levels in the hypophysectomized killifish, Fundulus heteroclitus, transferred from 50% seawater to fresh water. The effects of PRLs were parabolic, high doses of the hormones being less effective than low doses. When injected into seawater-adapted fry of the ayu, Plecoglossus altivelis, or into juvenile rainbow trout, Salmo gairdneri, adapted to 50% seawater, a dose-dependent increase in plasma sodium was observed. Chum PRL was 2-10 times more active than ovine PRL, and the effects in the ayu were also parabolic. An increase in plasma sodium also occurred when the PRLs were injected into the seawater-adapted eel, Anguilla japonica; the chum and ovine PRLs were equipotent, and hypercalcemia was also observed. In contrast, both chum and ovine PRLs were without effect on plasma sodium levels of chum salmon fry, either when injected into seawater-adapted fish kept in seawater or into fish subsequently transferred to fresh water. The absence of an effect of PRLs in chum salmon fry seems to be due, at least in part, to their good osmoregulatory ability during the period of seaward migration; effects of the exogenously administered PRLs may be compensated for by other hormones responsible for their hydromineral balance.

Adaptation, Physiological↗
PubMed1986

Immunocytochemical demonstration of pituitary cell types in the teleost Poecilia latipinna, by light and electron microscopy.

Using the unlabelled antibody method at the light microscope level, and the immunogold method at the electron microscope level, the distribution of the different adenohypophysial cells was demonstrated in the teleost Poecilia latipinna, by means of antisera to both teleostean and mammalian pituitary hormones and their subunits. Anti-salmon prolactin, but not anti-rat or -ovine prolactin, gave a specific staining of the acidophils of the rostral pars distalis (RPD), while anti-trout growth hormone (GH), but not anti-rat GH, stained similar but always separate cells in the proximal pars distalis (PPD). Antisera to the whole molecules of mammalian glycoprotein hormones stained the entire population of basophils in the PPD, but separate populations of gonadotrophs and thyrotrophs could be discriminated using anti-salmon gonadotrophin and anti-human thyrotrophin beta subunit. Antisera to ACTH (1-24) and (11-24) sequences, as well as beta-endorphin and met-enkephalin, stained the lead haematoxylin-positive cells of the RPD and pars intermedia (PI), whereas anti-alpha-MSH stained only the PI cells. Ultrastructural examination showed that these immunoreactivities were present in the same secretory granules, and were always greater in pale granules rather than electron dense granules. In the RPD, blebs of ACTH-immunoreactive cytoplasm were found to protrude through the gaps in the basement membrane into the neurohypophysis. The second "PAS-positive" cell type of the PI showed a strong cross-reaction with anti-salmon gonadotrophin, suggesting that it may produce a glycoprotein chemically related to the gonadotrophin(s).

Adrenocorticotropic Hormone↗
PubMed1986

Lack of glycosilation of pro-opiomelanocortin might account for the periodic acid-Schiff-negative reaction in ACTH cells of teleost fishes.

Unlike tetrapod ACTH cells, teleost ACTH cells do not react with the periodic acid-Schiff method (PAS). To find an explanation for this unique feature, chromatographic fractions obtained after filtration of pituitary extracts of Prochilodus platensis in Sephadex were immunologically analyzed. A high-molecular-weight protein which was identified as pro-opiomelanocortin (POMC) was detected. When this POMC was submitted to affinity chromatography in concanavalin binding, it was not detected. Furthermore, pituitaries incubated in media containing [3H]glucosamine or [3H]fucose did not incorporate these amino acids to the newly synthesized POMC. The results obtained strongly suggest an inability of the fish to glycosilate POMC, and this failure could account for the PAS-negative reaction in the ACTH cells.

Adrenocorticotropic Hormone↗
PubMed1987

The response of nucleus preopticus neurosecretory cells to ovarian pressure in the teleost, Clarias batrachus (Linn.).

The changes undergone by the neurosecretory cells of the nucleus preopticus (NPO) were investigated histologically and immunocytochemically following pressure stimulation of the ovaries in the teleost, Clarias batrachus. Application of intraovarian pressure of 10 mm Hg revealed marked hypertrophy of the cells, while pressures of 20 and 30 mm Hg resulted in hypertrophy and exhaustion of the cells. No changes were observed when the treatment was preceded by transection of the spinal cord. The results suggest the existence of a pressure-excitable neural pathway from the ovary capable of eliciting profound cytological changes in the NPO.

Animals↗
PubMed1987

Kinetics of T4 and T3 binding to plasma sites in salmonid teleost fish.

The relative affinities and maximum capacities of the classes of L-thyroxine (T4)- and 3,5,3'-triiodo-L-thyronine (T3)-binding sites in plasma of three species of salmonid teleost fish were determined by saturation analysis on miniature Sephadex G-25 columns at 20-21 degrees and kinetic data were analyzed by the multiligand, multisite LIGAND computer program. In plasma of rainbow trout (Salmo gairdneri) homologous ligand displacement indicated that both thyroid hormones (TH) bound to a minimum of two classes of saturable sites and at least one nonsaturable site. For T4 (n = 13) the relative site affinities were 0.61 +/- 0.08 (SEM) X 10(7) M-1 and 0.86 +/- 0.11 X 10(5) M-1 and the site capacities were 8.3 +/- 1.16 X 10(-7) M and 5.15 +/- 1.34 X 10(-5) M, respectively; for T3 (n = 14) the relative site affinities were 1.8 +/- 0.16 X 10(7) M-1 and 1.7 +/- 0.17 X 10(5) M-1 and the site capacities were 7.8 +/- 1.3 X 10(-7) M, respectively. The greater affinities of T3 than T4 for plasma binding sites would explain the lower proportion of free T3 than free T4 in trout plasma. Two-site models with comparable values for TH-binding parameters were determined for brook trout (Salvelinus fontinalis) and arctic charr (Salvelinus alpinus). The TH-binding parameters were uninfluenced by severe hemoglobin contamination of plasma, bleeding of fish 24 hr previously, or 2 weeks starvation. Heterologous ligand displacement (T4 displaced by T3 or T3 displaced by T4) on rainbow trout plasma suggested two low-affinity, high-capacity sites, one binding predominantly T4 and one binding predominantly T3: a high-affinity, low-capacity site binding T3 exclusively: and a high-affinity, low-capacity site binding T4 predominantly but also binding T3 with low affinity.

Animals↗
PubMed1987

Patterns and regulation of estradiol metabolism by hepatic microsomes from two species of marine teleosts.

Estradiol (E2) metabolites formed in vitro by microsomes from the marine teleosts winter flounder (Pseudopleuronectes americanus) and scup (Stenotomus chrysops) included at least seven products detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The least polar metabolite was shown to be estrone by chromatographic and mass spectrometric identity with authentic estrone. Chromatographic analyses coupled with dual-label experiments also indicated formation of the catecholestrogen 2-hydroxyestradiol (2-OH-E2), which was the most prominent metabolite determined by TLC. Analysis of microsomal E2 2-hydroxylase activity by measuring the specific release of 3H2O from [2-3H]E2 indicated that it is mediated by cytochrome P-450. E2 2-hydroxylase activity normalized to microsomal protein was lower in females than in males for microsomes from both mature scup and winter flounder. Activity normalized to liver weight or body weight in female winter flounder was also lower than that in males. However, activity normalized to cytochrome P-450 content did not show sex differences in either species. E2 2-hydroxylase activity per nanomole cytochrome P-450 was reduced in scup treated with beta-naphthoflavone, which induces the hydrocarbon hydroxylase cytochrome P-450E. Studies employing reconstituted P-450E and microsomes preincubated with polyclonal antibodies against P-450E confirmed that this isozyme does not catalyze E2 2-hydroxylase activity in microsomes. However, preliminary work with scup cytochrome P-450A suggests that it may be an E2 2-hydroxylase. The studies establish that catecholestrogen formation is prominent in fish liver and that it is sexually differentiated, but further investigation is required to define the catalysts as well as the significance and regulation of this function.

Animals↗
PubMed1987

A comparative immunocytochemical study of the gastro-entero-pancreatic (GEP) endocrine system in a stomachless and a stomach-containing teleost.

The gastro-entero-pancreatic (GEP) endocrine system of a stomach-containing and of a stomachless teleost, Sparus auratus and Barbus conchonius, respectively, are studied immunocytochemically using different antisera against mammalian hormones. Insulin-, glucagon-, somatostatin-, and pancreatic polypeptide (PP)-immunoreactive cells are identified in the endocrine pancreas of both species. Only the distribution of PP-immunoreactive cells differed strongly; in the principal islet of both fishes, few PP-immunoreactive cells are present, whereas in the smaller ones many of them are observed in S. auratus and none in B. conchonius. In the digestive tract of S. auratus 10 endocrine cell types can be distinguished: neurotensin-, secretin-, serotonin-, somatostatin-, and two types of substance P-immunoreactive cells exclusively in the stomach, and C-t-gastrin/CCK-, glucagon-, Met-enkephalin-, PP-, and only one type of substance P-immunoreactive cells in the intestinal epithelium. With the exception of substance P-immunoreactive cells, the other four intestinal endocrine cells, as well as an unspecific immunoreactive cell, can also be found in B. conchonius. Coexistence of glucagon- and PP-like immunoreactivity is observed in the pancreas of S. auratus and in the gut of B. conchonius. Pancreatic and gut endocrine cells showing only PP- or glucagon-like immunoreactivity are found, too.

Animals↗
PubMed1988

Melanin-concentrating hormone (MCH) immunoreactive hypophysial neurosecretory system in the teleost Poecilia latipinna: light and electron microscopic study.

Neurons containing immunoreactivity for melanin-concentrating hormone (MCH) were located in the brain of the teleost Poecilia latipinna by light microscopic (peroxidase antiperoxidase) and electron microscopic (immunogold) methods. Neuronal cell bodies were found in the tuberal hypothalamus, mostly within the nucleus lateralis tuberis, pars lateralis, containing MCH-immunoreactive granules up to 150 nm in diameter. From here bundles of immunoreactive fibers could be traced through the preoptic area as far forward as the olfactory bulb, and through the posterior hypothalamus up into the pretectal thalamus and midbrain. The main projection was, however, to the neurohypophysis, where MCH fibers were observed to form contacts with pituicytes, basement membranes around blood vessels, and the endocrine cells of the pars intermedia. Occasionally MCH-immunoreactive terminals were also seen near the corticotrophs of the rostral pars distalis. These results support the hypothesis that MCH may act as a systemic hormone, a central neurotransmitter, and a modulator of pituitary function.

Animals↗
PubMed1988

Immunocytochemical and ultrastructural characterization of the cell types in the adenohypophysis of Sparus aurata L. (teleost).

The structure and immunocytochemistry of the adenohypophysis of sexually mature male specimens of Sparus aurata (gilthead sea bream) were studied. The adenohypophysis was composed of rostral pars distalis (RPD), proximal pars distalis (PPD), and pars intermedia (PI). In the RPD the prolactin cells were organized into follicles which occupied a very reduced area as corresponds to that in saltwater fishes; the corticotropic cells were surrounding the pars nervosa branches and the prolactin follicles. The PPD showed somatotropic, gonadotropic, and thyrotropic cells. The somatotropic cells were isolated, clustered, or surrounding the pars nervosa branches. Only one polymorphic cell type of gonadotropic cells was found in the PPD ventral and dorsal areas and around the PI. The PI was composed mainly of melanotropic cells and a PAS-positive cell layer adjacent to the neurohypophysis. The ultrastructure of the presumptive endocrine cells was reported and their distribution was discussed in relation to those of other teleosts.

Adrenocorticotropic Hormone↗
PubMed1988

Development of sensitivity to maturation-inducing steroids in the oocytes of the daily spawning teleost, the kisu Sillago japonica.

The responsiveness of oocytes to 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-diOH-P), 17 alpha-hydroxyprogesterone (17 alpha-OH-P), and silver carp gonadotropin (GtH) was examined in vitro at seven different times of the day (0100, 0500, 0800, 1000, 1300, 1700, 2100 hr) in a daily spawning teleost, the kisu Sillago japonica. 17 alpha,20 beta-DiOH-P, 17 alpha-OH-P, and GtH were effective in inducing germinal vesicle breakdown (GVBD) and ovulation in kisu oocytes. However, the oocytes of the kisu responded to these hormones during a limited time of the day (2100-1000 hr), and their responsiveness to steroids and GtH was different with the sampling time. After a 20-hr incubation, the oocytes with yolk globules were classified into four stages based on their responses (GVBD) to GtH and steroids. Stage A, the oocytes did not respond to either GtH or steroids. These unresponsive oocytes were observed in all the ovaries examined. Stage B, GVBD could be induced by GtH but not by steroids. These oocytes were found in the ovaries collected from 2100 to 1000 hr. Stage C, GVBD could be induced by both GtH and steroids. These oocytes were found only between 0800 and 1000 hr. Stage D, oocytes spontaneously underwent GVBD without hormones. These oocytes were found between 1300 and 1700 hr. Ovulated oocytes were found in the ovaries collected during 1700-0500 hr. These results indicate that the kisu possesses a daily rhythm in oocyte development from stage A to stage D, and that the sensitivity to 17 alpha,20 beta-diOH-P appears at stage C.(ABSTRACT TRUNCATED AT 250 WORDS)

17-alpha-Hydroxyprogesterone↗
PubMed1989

Effect of a teleost GnRH analog on steroidogenesis by the follicle-enclosed goldfish oocytes, in vitro.

The influence of an agonist analog of teleost GnRH [(D-Arg6, Trp7, Leu8, Pro9-NEt)-GnRH; tGnRH-A] on steroidogenesis was studied in prophase-I arrested, follicle-enclosed, goldfish oocytes in vitro. Incubation of the follicles with carp gonadotropin (GtH) significantly increased production of 17 alpha-hydroxyprogesterone (HP) and testosterone following 24 hr of incubation in vitro. Concomitant incubation with tGnRH-A (10(-7) M) significantly attenuated the dose-related increase in GtH-induced testosterone production, but was without effect on the GtH-induced HP level. Time course studies indicated that tGnRH-A exerted its maximum inhibitory action on the GtH-induced testosterone production during the initial 8 hr of incubation in vitro. The inhibition of GtH-induced testosterone production by tGnRH-A was dose dependent with an ED50 of 1.39 +/- 2.88 nM. A significantly higher testosterone level was obtained in the incubation media containing HP as substrate; concomitant treatment with tGnRH-A reduced the conversion of HP to testosterone. The incubation media also contained low, but measurable levels of 17 alpha-hydroxy-20 beta-dihydroprogesterone (DHP), which increased in the presence of 3-isobutyl-methyl-xanthine; lower levels of DHP were obtained in the groups incubated with tGnRH-A. In view of our present findings and previous observations concerning inhibitory effects of tGnRH-A on the progestogen and GtH-induced reinitiation of meiosis in the follicle-enclosed goldfish oocytes (H. R. Habibi, G. Van Der Kraak, E. Bulanski, and R. E. Peter, Amer. J. Physiol. 255, R268-R273 (1988] the influence of testosterone on the GtH- and DHP-induced meiosis in vitro was also studied. Testosterone (1 micrograms/ml) enhanced both GtH- and DHP-induced oocyte meiosis in the goldfish oocytes. Testosterone alone was also found to significantly increase oocyte meiosis in the goldfish oocytes in a dose-related fashion. The present findings demonstrate an inhibitory effect of a GnRH agonist on GtH-induced testosterone production in goldfish oocytes and suggest that tGnRH-A might influence oocyte meiosis in part by influencing steroidogenesis.

Animals↗
PubMed1989

Purification and properties of pituitary gonadotropic hormone from Indian teleosts: freshwater murrel (Channa punctatus) and carp (Catla catla).

Gonadotropic hormone (GtH) from the pituitaries of two widely different Indian teleosts, a freshwater murrel and a carp, was purified by solvent fractionation, Sephadex G-100 gel filtration, affinity chromatography on concanavalin A-Sepharose (Con A-Sepharose), and immunoaffinity chromatography. Elution profile from gel filtration showed three peaks in both cases, peak I and II were clearly separated in murrel but not in carp. Peak II demonstrated strong GtH activity in both murrel and carp but this activity could also be detected in peak I and III of carp. TSH activity was restricted to peak I in murrel and was distributed in all three peaks in carp. Chromatography on Con A-Sepharose was useful in harvesting carp glycoprotein hormones, but gave no advantage in murrel. A pure homogenous GtH from murrel and carp could be obtained by using immunoaffinity chromatography. Polyacrylamide gel electrophoresis of murrel and carp GtH showed a single discrete band. Determination of molecular weight (MW) by Sephadex G-100 gel filtration indicated that murrel and carp GtH were 42,000 and 40,000 Da, respectively. SDS-polyacrylamide gel electrophoresis of murrel and carp GtH revealed two dissimilar subunits, alpha and beta. MW of murrel and carp alpha-subunits were 18,000 and 16,000 while those of beta were 27,000 and 26,000 Da, respectively. Comparison of electrophoretic patterns of murrel and carp GtH alpha and beta with ovine LH subunits reveal distinct teleostean GtH subunits.

Animals↗
PubMed1989

A new member of the gonadotropin-releasing hormone family in teleosts: catfish gonadotropin-releasing hormone.

Two forms of immunoreactive gonadotropin-releasing hormone (GnRH) were detected in extracts of brain-pituitary tissue from the African catfish, Clarias gariepinus. Catfish I GnRH eluted first from reverse-phase HPLC and was present in larger amounts compared with catfish II GnRH. Chromatographic and immunological studies with four antisera provide evidence that catfish I GnRH is unique compared with identified GnRHs from mammal, chicken, salmon, and lamprey. Catfish II GnRH elutes in the same position as chicken II GnRH and the forms cannot yet be distinguished. GnRHs extracted from female and male catfish tissue appear to be similar in terms of the number of peaks eluted, elution position, quantity, and cross-reactivity with the antisera. The results of the HPLC and radioimmunoassay studies suggest that catfish I GnRH is likely to be 10 amino acids in length, and have an amide at the C terminus similar to the other family members. In addition, catfish I GnRH is probably different in the 5 to 10 amino acid region compared with mammalian GnRH. Finally, catfish I GnRH is likely to have a lysine or arginine residue as it is the most hydrophilic family member. The lack of the salmon form of GnRH and the presence of a unique GnRH form constitute another example of the considerable evolutionary variation that has occurred in the catfish family compared with other teleosts.

Animals↗