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Evidence for 5-HT-containing intrinsic neurons in the teleost intestine.

The stomach and small intestine of seven species of teleost fish were examined with both fluorescence and ultrastructural histochemical techniques for demonstrating monoamines. Fluorescence histochemistry demonstrated large numbers of yellow fluorescent neurons in the myenteric plexus of the intestine, but not the stomach, of four species. Varicose fibres arising from these neurons ran in the myenteric plexus, circular muscle layer and submucosa. Ultrastructural histochemistry of tissue fixed with a modified chromaffin reaction showed that some neurons in the myenteric plexus contained biogenic amines in vesicles of 40--100 nm diameter. It is concluded that 5-HT-containing neurons are present in the intestine of some teleost species.

Animals↗

Immunocytochemical identification of thyrotropin (TSH)-producing cells in pituitary glands of several species of teleosts with antiserum to human TSH beta subunit.

A rabbit antiserum raised against human thyrotropin (TSH) beta subunit (anti-human beta-TSH) was used to identify TSH cells in the pituitaries of five species of teleosts by means of the unlabeled antibody peroxidase-antiperoxidase (PAP) technique. The anti-human beta-TSH reacted only with polygonal basophils located in the rostral pars distalis (RPD) in the pituitaries of sexually immature eel (Anguilla japonica). In the pituitaries of medaka (Oryzias latipes) and white-edged rockfish (Sebastes taczanowskii), this serum reacted specifically with only one cell-type located in the antero-dorsal proximal pars distalis (PPD) and antero-ventral PPD, respectively. In the pituitaries of two species of salmonids, sexually immature rainbow trout (Salmo gairdneri) and sexually mature chum salmon (Oncorhynchus keta), the anti-human beta-TSH bound to the polygonal basophils in the ventral PPD adjacent to the RPD. Thus, the present immunocytochemical study identified polygonal basophils located in the RPD or PPD as TSH cells in five species of teleosts.

Anguilla↗

Optic tract cells projecting to the retina in the teleost, Pantodon buchholzi.

Horseradish peroxidase was employed to trace retino-fugal and retino-petal connections in the teleost fish, Pantodon buchholzi. Most of the reciprocal connections found were within the range also observed in previously studied species of teleosts. Of particular interest is the discovery of cells located within the optic tract and projecting to the retina. These neurons were investigated electron microscopically.

Animals↗

The relationship between the ionic composition of the environment and the secretory activity of the endocrine cell types of Stannius corpuscles in the teleost Gasterosteus aculeatus.

The corpuscles of Stannius of threespined sticklebacks contain two glandular cell types of presumed endocrine nature. To elucidate the function of both cell types the secretory activity of the cells was studied in fully adapted seawater and freshwater fishes and in specimens transferred from sea water to fresh water or adapted to media of various ionic composition. The secretory activity was established, in tissue sections and freeze-etch replicas, by estimating the volume of the nuclei, the density of the nuclear pores, and the frequency of exocytotic phenomena. The type-1 cells, ultrastructurally comparable to the predominant or only cell type described in many other teleosts, are more active in sea water than in fresh water. The activity of the type-2 cells, whose ultrastructural appearance is known only for salmonids and eels, is higher in fresh water. Transfer of seawater fishes to fresh water results in reduction of type-1 cells and activation of type-2 cells. The factors responsible for these changes were analyzed by exposure of fishes to solutions of various salts in fresh water and to artificial sea water with a reduced content of one of its components. The high activity of type-1 cells in sea water proved to be related to the high calcium content of this medium. These cells probably produce a substance comparable to hypocalcin, the endocrine factor isolated from the Stannius corpuscles of some other teleost species. The high activity of type-2 cells in fresh water appeared to be connected with the low sodium and potassium levels of this medium. Type-2 cells possibly produce a hitherto unknown hormone involved in the control of sodium and/or potassium metabolism.

Animals↗

Electron microscopic studies on the development of the chorion of the viviparous teleost Dermogenys pusillus (Hemirhamphidae).

Eight stages in the oogenesis of Dermogenys pusillus were selected in order to demonstrate the formation of the egg membrane. In young oocytes (stages 1 and 2) the contact between oocyte and follicular cells is rather close. During stage 3 microvilli arise from the oocyte, and the follicular cells protrude lobopodia-like cell processes. When the microvilli have become arranged more regularly, the homogeneous material of the zona radiata externa is deposited between them (stage 4). During the subsequent stage (5) the inhomogeneous zona radiata interna appears. It attains its greatest thickness and its characteristic fine structure during stage 6. It then consists of cross-banded fibrillae and lamellae. Later on (stage 7) the egg membrane flattens, and finally it is a compact chorion consisting perhaps of very fine fibrils. Because of this development and of its comlex fine structure the egg membrane of Dermogenys like that of Zoarces resembles that of oviparous teleosts but remains consideribly thinner. The eight stages of oogenesis in Dermogenys are compared with those described by Arndt (1956) in several limnetic teleosts.

Animals↗

Ultrastructure of the adenohypophysis in the teleost Poecilia latipinna.

In the sailfin molly, Poecilia latipinna, seven morphological endocrine cell-types could be distinguished with the electron microscope. Each of these was identified with one of the seven cell-types distinguished with the light microscope, to most of which endocrine functions have previously been allocated. Corticotrophs and prolactin cells form the rostral pars distalis, and the proximal pars distalis consists of an outer layer of gonadotrophs and an inner zone containing growth hormone cells and thyrotrophs. The pars intermedia contains two cell-types, of uncertain function. Stellate cells (interstitial cells) occur throughout the adenohypophysis, but are most numerous and prominent in the rostral pars distalis. The inner proximal pars distalis contains a cell-type not previously distinguished in this species with the light microscope, the Z-cell, which could be aminergic. The ultrastructural features of each cell-type are described in detail, and discussed in comparisons with the homologous cells described in other teleosts. There is good agreement for different teleosts in the ultrastructural details of each cell type.

Adrenocorticotropic Hormone↗

Immunohistochemical demonstration of calbindin-D 28K (CABP28K) in the spinal cord motoneurons of teleost fish.

The distribution and localization of the calcium-binding protein, calbindin-D 28K (CaBP28K), in the spinal cord motoneurons of larvae of the teleost fish, Apteronotus leptorhynchus (Gymnotidae) and Pollimyrus isidori (Mormyridae), and in the adult goldfish, Carassius auratus (Cyprinidae), were determined by means of immunohistochemistry. Sections of whole larvae and goldfish spinal cord were reacted with a polyclonal antibody to rat renal CaBP28K. CaBP28K was located by the PAP technique (Sternberger). It was found in the soma, dendrites, axons and axon terminals of spinal motoneurons but not in those of electromotoneurons of Apteronotus leptorhynchus, whereas it occurred in both motoneurons and electromotoneurons of the larval electric organ of Pollimyrus isidori. In these species CaBP28K was also present in the electromotoneuron axon terminals that make synaptic contacts with the pedicles of the electrocytes. In adult Carassius auratus, CaBP28K was found in the soma, dendrites and axons of certain spinal motoneurons. The results indicate that, in teleosts, the motoneurons containing CaBP28K may represent a well-defined population within the spinal cord; the role of this protein in these cells remains to be determined.

Animals↗

Mitochondria-rich (chloride) cells in the gill epithelia from four species of stenohaline fresh water teleosts.

The mitochondria-rich (chloride) cells have been found to be present in the gill epithelia of four species of stenohaline fresh water teleosts. The cytoplasm of these chloride cells contains an extensive network of cytoplasmic tubules which communicate with intercellular spaces bordering the lateral and basal cell surfaces. Numerous vesicles with fairly electron-dense interiors are also present in the apical cytoplasm of chloride cells. The apical surface of a chloride cell forms an apical pit, but the lumen of the pit does not appear to be in continuity with the interior of the apical vesicles and tubules inside the cell. When Carassius auratus were kept in 100, 200, 300, and 400 mOsm-diluted sea water for a month, no appreciable changes occurred in the number and fine structure of the chloride cells, except for a dilation of the apical vesicles and a slight decrease in diameter of the cytoplasmic tubules in these cells in the fishes kept in 300 and 400 mOsm. These results suggest that chloride cells may be a rather common occurrence in the gill epithelia of stenohaline fresh water teleosts, and may function in ion transport in these fishes in fresh water environments.

Animals↗

Capillary distribution and metabolic histochemistry of the lateral propulsive musculature of pelagic teleost fish.

Metabolic and vascular adaptation of teleost lateral propulsive musculature to an active mode of life was investigated in four pelagic teleosts (mackerel, yellowtail scad, pilchard and Australian salmon). Histochemical profiles and capillarisation data of the red and white muscle were compared to those of less active demersal species. Pelagic white muscle stained positively for the aerobic enzymes succinate dehydrogenase and NADH diaphorase, and had both subsarcolemmal and intermyofibrillar mitochondria which corresponded to the loci of the histochemical stain. Subsarcolemmal mitochondria tended to be localised close to capillaries. In contrast, white muscle from demersal species was unstained for the same enzymes and was devoid of mitochondria. Red muscle of all species had abundant mitochondria and stained intensely for aerobic enzymes. Capillarisation was quantified by determining the percentage of fibres surrounded by a given number of peripheral capillaries, mean fibre diameter, mean number of peripheral capillaries, capillary: fibre ratio and sharing factor where appropriate. Red muscle of mackerel, Australian salmon, pilchard and scad are better vascularised than red muscle of the flathead having 153, 200, 242, 291 and 309 microns 2 of cross-sectional fibre area per peripheral capillary, respectively. White muscle of mackerel, pilchard and scad are better vascularised than white muscle of the Australian salmon and flathead having 2040, 3367, 4992, 9893 and 10,469 microns 2 of cross-sectional fibre area per peripheral capillary, respectively. Red muscle of Australian salmon had distinct regional variation. Deep red muscle was found to be more highly vascularised (4.2 peripheral capillaries per muscle fibre) than lateral red muscle (1.9 peripheral capillaries per muscle fibre). Red muscle of the other species was less heterogeneous. White muscle capillarisation was slightly variable in all species. It is concluded that the white muscle of the pelagic species studied is functionally and structurally adapted for sustained aerobic activity with relatively abundant mitochondria being preferentially situated close to the source of gas and metabolite exchange.

Adenosine Triphosphatases↗

The distribution of aminergic neurones and their projections in the brain of the teleost, Myoxocephalus scorpius.

Fluorescent histochemistry was carried out on the brain of the teleost Myoxocephalus scorpius to show the distribution of monoaminergic neurones and their projections. Posterior to the obex of the fourth ventricle, at the junction of the spinal chord and medulla, there is an unpaired dorsal nucleus of catecholaminergic cells. A second group of catecholaminergic perikarya are scattered lateral to the vagal and glossopharyngeal motor nuclei. Both groups of aminergic cells contribute to a tract which crosses the fourth ventricle at the obex and runs along the lateral wall of the medulla towards the diencephalon. At the level of the isthmus there is a lateral nucleus composed of large catecholaminergic cells with prominent fluorescent axons and its possible homology with the locus coeruleus is considered. Medially, in the same region a nucleus of serotonergic neurones lies between the paired tracts of the fasciculus longitudinalis medialis. In the diencephalon there are three paraventricular nuclei, the nuclei recessus posterioris and lateralis and the paraventricular organ pars anterior. Ventral to the lateral recess there is a further nucleus less closely associated with the ependyma. The distribution of fluorescent fibres is described and the dispositions of the aminergic nuclei compared to those of other teleosts.

Animals↗

Immunocytochemical localization of a galanin-like peptidergic system in the brain and pituitary of some teleost fish.

Immunostaining of brain and pituitary sections of teleost fishes (eels, salmonidae, cyprinidae, gourami, sculpin, mullet) with anti porcine galanin (GAL) revealed the presence of immunoreactive (ir) perikarya and a rich network of fibers. Ir-perikarya were located rostrodorsally to the recessus preopticus, and in the posterior tuberal hypothalamus. Ir-fibers were abundant in basal telencephalon and around diencephalic ventricular recesses but never contacted their lumen. Furthermore, they were observed in basal hypothalamus, brainstem and ventral medulla. Ir-fibers passed along corticotropic (ACTH), gonadotropic cells and somatotropes (GH cells) in eel and trout pars distalis, but rarely ended in caudal neurohypophysis. In goldfish pituitary ir-fibers occurred in neural digitations and among different cell types which however did not contain a GAL-like peptide. The relation GAL fibers/GH cells appeared more evident in species with a high growth rate. The other species showed a similar distribution of brain GAL. In eels and trout, ir-perikarya were not observed in areas containing somatostatin, GH- and ACTH-releasing factor, and ACTH-like perikarya, suggesting that GAL did not coexist with these peptides. The widespread distribution of a GAL-like peptide in teleost brain suggests that it could play a role of neurotransmitter and/or neuromodulator and regulate the secretion of adenohypophysial hormone(s).

Animals↗

Fluorescent labelling of Na+, K(+)-ATPase in intact cells by use of a fluorescent derivative of ouabain: salinity and teleost chloride cells.

Anthroylouabain, a fluorescent derivative of ouabain, was used to localize Na+, K(+)-ATPase in transport epithelia of two species of teleosts. Exposure of the opercular membrane of seawater-adapted tilapia (Oreochromis mossambicus) and the jaw skin of the long-jawed mudsucker (Gillichthys mirabilis) to a 2 microM anthroylouabain solution resulted in the appearance of cells stained bright blue. These were deemed to be chloride cells by their large size, distinct morphology and co-localization of DASPEI fluorescence, a mitochondrial stain. Addition of ouabain (1 mM final concentration) greatly decreased anthroylouabain fluorescent staining of chloride cells of seawater-adapted fish. Exposure of opercular membranes from freshwater tilapia to 2 microM anthroylouabain did not result in significant staining. Anthroylouabain is therefore a useful vital stain for localizing Na+,K(+)-ATPase in chloride cells of seawater-adapted teleosts, and may be useful for fluorescent labelling of ouabain-sensitive Na+,K(+)-ATPase in other tissues and species.

Animals↗

Distribution of PACAP (pituitary adenylate cyclase-activating polypeptide)-like and helospectin-like peptides in the teleost gut.

Pituitary adenylate cyclase-activating polypeptide (PACAP) and helospectin are two vasoactive intestinal polypeptide (VIP)-related neuropeptides that have recently been demonstrated in the mammalian gut; the aim of this study was to reveal their occurrence and localisation in the gastrointestinal tract, swimbladder, urinary bladder and the vagal innervation of the gut of teleosts, using immunohistochemical methods on whole-mounts and sections of these tissues from the Atlantic cod, Gadus morhua and the rainbow trout, Oncorhynchus mykiss. Both PACAP-like and helospectin-like peptides were present in the gut wall of the two species. Immunoreactive nerve fibres were found in all layers but were most frequent in the myenteric plexus and along the circular muscle fibres. Immunoreactivity was also demonstrated in nerves innervating the swimbladder wall, the urinary bladder and blood vessels to the gut. Immunoreactive nerve cell bodies were found in the myenteric plexus of the gut and in the muscularis mucosae of the swimbladder. In the vagus nerve, non-immunoreactive nerve cells were surrounded by PACAP-immunoreactive fibres. Double staining revealed the coexistence of PACAP-like and helospectin-like peptides with VIP in all visualized nerve fibres and in some endocrine cells. It is concluded that PACAP-like and helospectin-like peptides coexist with VIP in nerves innervating the gut of two teleost species. The distribution suggests that both PACAP and helospectin, like VIP, are involved in the control of gut motility and secretion.

Amino Acid Sequence↗

Serotonin-like immunoreactivity in the epidermal club cells of teleost fishes.

The present immunocytochemical study concerns the distribution of serotonin in the epidermis of three species of teleost fish. Serotonin-like immunoreactivity was found in the club cells of Heteropneustes fossilis and Carapus acus but not in those from the sea eel Conger conger. This study is the first immunocytochemical identification of serotonin in the club cells of teleost epidermis. By comparing data from the literature (Zaccone et al. 1986, 1987, 1988) regarding the occurrence of serotonin and GRP/bombesin in the exocrine sacciform gland cells of piscine skin, it is worthy mentioning here that the serotonin contained in the club cells of the species studied may have the ability to affect the pheromonal or other possible functions of these cells. The presence of serotonin in these systems has been correlated with the capacity of the exocrine glands of fish skin to secrete, ectopically, amine messengers in contrast to those produced eutopically i.e. in the neuron-paraneuron system in some vertebrates (Fujita et al. 1988).

Animals↗

Histochemical localization of acetylcholinesterase in the cerebellum and optic tectum of four freshwater teleosts.

A histochemical study has been carried out on the localization of acetylcholinesterase (AChE) in the cerebellum and optic tectum of four species of freshwater teleosts. AChE distribution in the cerebellar cortex of teleosts shows differences among the species examined and, in the trout, also differences between different cerebellar areas. This uneven kind of enzyme distribution corresponds to a similar variety of AChE patterns noticed in other vertebrates, especially mammals. AChE distribution in the optic tectum shows a prevalent pattern characterized by precise laminar distribution of enzymatic activity which is alternatively strong, weak or absent in the different tectal layers. The results suggest that most of sensitive imput and many systems of stimuli propagation may be mediated by cholinergic mechanisms in the optic tectum of telecosts.

Acetylcholinesterase↗

Localization of growth hormone-releasing factor-like immunoreactivity in the hypothalamo-hypophysial system of some teleost species.

An antiserum to growth hormone-releasing factor (GRF) 1-44 was applied on brain and pituitary sections of nine teleost species. Immunoreactive (ir) perikarya were demonstrated in parvo- and magnocellular portions of the preoptic nucleus (PON) and occasionally in the nucleus lateralis tuberis. The two tracts originating in the PON ran ventro-laterally toward the optic chiasm and then caudally in the basal hypothalamus. In the pars distalis (PD) of the eel, carp, goldfish and salmonids, GRF-ir fibers did not enter the rostral PD and few fibers passed close to somatotropes. In Myoxocephalus and Mugil, a variable number of ir-fibers passed close to cells of the rostral and proximal PD. In the neurointermediate lobe, GRF-ir fibers were located exclusively in the neural tissue of the eel and trout. In goldfish, carp and Myoxocephalus, GRF-ir fibers entered the intermediate lobe. This antiserum also labeled corticotrops and, to a lesser extent, melanotrops in the pituitary of cyprinids. A variable number of perikarya contained both GRF and vasotocin in the PON of the eel. In all teleost species studied so far, the distribution patterns of GRF are different, and the function of the various adenohypophysial cell types appears to be differently modulated, according to the variable distribution of GRF in the pituitary.

Anguilla↗

The mechanical properties of polyneuronally innervated, myotomal muscle fibres isolated from a teleost fish (Myoxocephalus scorpius).

Single or small bundles of fibres were isolated from the abdominal myotomes of the sculpin Myoxocephalus scorpius, a teleost with a polyneuronal pattern of fast muscle innervation. Fibres responded to a supra-threshold stimulus with an all-or-none twitch. Tetanic fusion frequency at 3 degrees C was 40-60 Hz, and the twitch tetanus ratio 0.70. Maximum isometric tension was 281 kN m-2. Similar isometric contractile properties were obtained from the focally innervated fast muscle fibres of another teleost, the eel, Anguilla anguilla. The response of sculpin fibres to stretch during tetanus was similar to that reported for frog twitch fibres. A 5% stretch of 25-50 ms duration increased force to 1.4 Po which decayed to a steady level 5-10% above that of a control tetanus. The force-velocity relationship was also studied. Maximum contraction velocity was 4.75 Ls-1. Force-velocity data were not adequately described by a simple hyperbola. Alternative methods of curve fitting have been explored and discussed.

Anguilla↗

Morphological evidence for a direct neuroendocrine GABAergic control of the anterior pituitary in teleosts.

The anterior pituitary of teleosts is unique among vertebrates in receiving a direct innervation which represents the morphological support of the neuroendocrine control of pars distalis functions. The participation of GABAergic fibers in this innervation was studied by means of immunocytochemistry at the light and electron microscopic levels, using antibodies against GABA. Immunoreactive fibers, characterized by the presence of small clear and dense cored vesicles, were detected in all parts of the gland. Immunopositive terminals were found in close, sometimes synaptic-like, contact with most glandular cell types in the anterior lobe. The data strongly suggest that in teleosts, as in mammals, GABA is involved in the neuroendocrine control of anterior pituitary functions.

Animals↗