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At least 19 recordsLinked to original sources

PubMed1984

Organization of interstitial tissue in the testis of the salamander Necturus maculosus (Caudata:Proteidae).

In Necturus maculosus the organization of the interstitial tissue varies according to the stage of spermatogenesis. Leydig cells at various stages of differentiation and myoid cells are always present in this tissue. The Leydig cells are undifferentiated at all phases of germ cell activity and only hypertrophy following spermiation and degeneration of Sertoli cells. These Leydig cells are structurally analogous to mammalian Leydig cells. They do not form part of the lamina propria of the seminiferous lobules and hence cannot be referred to as lobule-boundary cells previously described in the urodele testis (Lofts, '74). When the Leydig cells hypertrophy, numerous unmyelinated axons appear in the interstitial tissue. These axons, often devoid of Schwann-cell cytoplasm, occur in close proximity to Leydig cells. Because the levels of both Substance P and neurotensin increased in the testis of Necturus maculosus as Leydig cells differentiated, we concluded that these neural elements may regulate Leydig-cell function locally, through the release of neuropeptides.

Animals↗

PubMed1992

Identification of an androgen receptor in the zonal testis of the salamander (Necturus maculosus).

The testis of the salamander, Necturus maculosus, is advantageous for studying biochemical changes during spermatogenesis because germ cells and associated Sertoli and Leydig cells are topographically separated by stage of development. Using extracts of staged tissue samples and [3H]testosterone (T) in a standard binding assay, followed by Sephadex LH-20 or DNA-cellulose chromatography to separate free and bound steroid, we have identified a T-binding protein having physicochemical characteristics of a classical androgen receptor (AR): high affinity (Kd = 10(-9) M), limited capacity (Bmax) = 10(-10) M or 350 fmol/g tissue) and androgen specificity (T = 5 alpha - dihydrotestosterone greater than progesterone = corticosterone greater than estradiol). AR was present in nuclear extracts, where greater than 80% of binding sites were occupied by endogenous ligand, but was not detectable in cytosol. On linear sucrose gradients, nuclear AR sedimented at 3-4 S in both low and high ionic-strength buffers and, by this and other criteria, was distinguishable from the nonreceptor androgen binding protein (ABP) of the same species. The diffuse distribution of AR in germinal and nongerminal (glandular) tissues at all developmental stages is consistent with a dual localization in Sertoli cells and Leydig cells, as previously reported in mammals, and further suggests a regulatory role of androgen throughout spermatogenesis.

Androgen-Binding Protein↗

PubMed1980

Electron microscopic and autoradiographic studies on vitellogenesis in Necturus maculosus.

Electron microscope studies on Necturus maculosus oocytes ranging in size from 1.1-1.5 mm in diameter indicate the primary proteinaceous yolk to arise within structures referred to in other amphibian oocytes as yolk precursor sacs or bodies. The origin of these yolk precursor sacs appears to result from the activity of the Golgi complexes which form multivesicular and granular-vesicular bodies, the limiting membrane of which is at times incomplete. During differentiation, the yolk precursor sacs contain small vesicles similar in size to Golgi vesicles, larger vesicles similar to vesicular elements of the agranular endoplasmic reticulum and, on occasion, a portion of a mitochondrion. The interior of these sacs becomes granular, perhaps by a dissolution of the components just described, and soon becomes organized into a crystalline configuration. In oocytes 2.0-2.5 mm in diameter, an extensive micropinocytotic activity begins, continues throughout vitellogenesis, and constitutes the primary mechanism for the formation of secondary yolk protein. Numerous coated and smooth-surfaced vesicles, as well as electron-dense and electronlucent ones, fuse in the cortical ooplasm to form progressively larger yolk platelets.

Animals↗

PubMed1988

Ultrastructure of taste cells and synapses in the mudpuppy Necturus maculosus.

Taste buds in the mudpuppy Necturus maculosus were examined with electron microscopy. Three cell types (dark, light, and basal) were identified and reconstructed from serial thick sections. Dark and light cells extend from the basal lamina to the surface of the tongue. The apical process of the dark cells was usually quite lamellar when viewed in cross section, in contrast to light cells, whose apical process appeared more cylindrical. Basal cells are situated at the base of the bud and do not extend processes to the surface of the tongue. The cytoplasm of basal cells contains numerous clear and dense-cored vesicles. Small, spinelike processes (2-3 microns in length) project outward from the basal cells into the cytoplasm of the surrounding tast receptor cells. Morphologically, basal cells in mudpuppy taste buds resemble Merkel cells. Unmyelinated afferent nerve fibers enter the taste bud at the base and course through the lower portion of the bud. Synapses were found between taste receptor cells and nerve fibers, between basal cells and nerve fibers, and between basal cells and taste receptor cells. Over 65% of the synapses observed in the mudpuppy taste bud involved the basal cell. These findings suggest that basal cells play some role in chemosensory signal processing or integration of the taste response.

Animals↗

PubMed1968

Organization and activity in the pre- and postovulatory follicle of Necturus maculosus.

The established follicle envelope of Necturus maculosus consists of a layer of follicle cells (granulosa) surrounding the developing oocyte, a layer of theca comprised of connective tissue cells, fibers, and matrix, and a layer of serosal cells. The changes in shape and fine structure of these layers during differentiation accompanying oogenesis are described. The cells and capillaries of the follicle envelope are engaged in an extensive pinocytotic activity, the details of which are described. We used cytochemical techniques to analyze the activity of the follicle envelope with respect to lipid accumulation and alkaline phosphatase activity. Radioautographic results indicate that cells of the follicle envelope are capable of incorporating tritium-labeled uridine and amino acids at certain times during oocyte growth. A comparative analysis was made of the soluble proteins in follicle envelopes isolated from immature oocytes and of those in follicle envelopes isolated from nearly mature oocytes and in postovulatory follicles. After the oocyte is ovulated, the cells of the follicle envelope are converted into a postovulatory follicle. The cells of the postovulatory follicle undergo further differentiation resulting in their becoming actively engaged in the formation of a secretion, the details of which are described at the electron microscope level. Analysis of the postovulatory follicle by thin-layer chromatography and cytochemistry demonstrated the presence of a wide variety of lipid substances and the possible presence of steroid. That the postovulatory follicle may be engaged in steroid biosynthesis is also suggested by studies involving the demonstration of 3 beta-hydroxysteroid dehydrogenase activity with cytochemical techniques applied to frozen sections and to soluble proteins separated by gel electrophoresis.

Animals↗

PubMed1984

Fine structure of the abdominal epidermis of the adult mudpuppy, Necturus maculosus (Rafinesque).

The ventral epidermis of adult Necturus maculosus has been studied using electron and light microscopy. Many larval characteristics of amphibian epidermal structure are retained in adult Necturus. The epidermis is a stratified epithelium consisting of four cell layers and five cell types. Major differences compared with other adult amphibians are: the absence of a well defined moulting cycle together with an apparently diminished synthetic and mitotic activity in the stratum germinativum; an outermost cell layer (stratum mucosum) that is unkeratinized and appears to synthesize a mucous layer; and numerous large club-shaped Leydig cells which span the epidermis between the cells of the stratum germinativum and stratum mucosum. The apical region of the stratum granulosum and stratum mucosum cells shows evidence of extensive synthesis. The stratum mucosum appears to be involved in the secretion of vesicular contents onto the outermost surface of the epithelium. The external surfaces of the stratum mucosum cells possess numerous microridges which are supported by an intricate network of cytofilaments in the apical region of these cells. The significance of these features is discussed in relation to the physiology and ecology of this species.

Abdomen↗

PubMed1985

Control of the heart in the mudpuppy, Necturus maculosus.

The cardiac innervation of the mudpuppy, Necturus maculosus, was investigated using in vivo recording of blood pressure and heart rate, a perfused heart preparation, isolated paced ventricular and atrial strip preparations, Falck-Hillarp fluorescence histochemistry and measurements of plasma concentrations of catecholamines. The heart of Necturus receives a 'classical' vagal inhibitory innervation by cholinergic fibres which act via muscarinic cholinoceptors. In addition, there appears to be an atropine-resistant inhibitory component in the vagal innervation of the heart. This component is best seen at higher frequencies of vagal stimulation (10-20 Hz), but the mechanism behind this response is not known. There is no direct adrenergic innervation of the ventricle or atria, but by use of Falck-Hillarp fluorescence histochemistry, adrenergic fibres have been demonstrated at the atrio-ventricular border, particularly within the intracardiac ganglion. The histochemical evidence is consistent with a modulation of the ganglionic transmission in the vagal pathways by adrenergic neurons and small intensely fluorescent (SIF) cells, but physiological evidence is wanting. An adrenergic control of the heart via circulating adrenaline may occur during 'stress', when the plasma level of adrenaline is high enough to affect cardiac performance.

Acetylcholine↗

PubMed1977

The central projections in the retina in Necturus maculosus.

The projections of the retina in Necturus maculosus were studied by injecting radioactive proline into one eye. Labeling was seen in both the contralateral and ipsilateral diencephalon and tectum. The contralateral fibers are divided into three major tracts: the marginal, axial, and basal. The ipsilateral fibers separate into a marginal and an axial optic tract. The contralateral and ipsilateral axial optic tracts have a similar distribution. The contralateral and ipsilateral marginal optic tracts projecting to the diencephalon also have a similar distribution. However, in the tectum the ipsilateral marginal optic tract ends in the anterior third while the contralateral extends almost the entire length of the tectum. The retinotectal ipsilateral projection ends in clumps as has been described in other vetebrates. A direct ipsilateral retinotectal projection has not been described in any other amphibian.

Animals↗

PubMed1992

Chemosensory responses in isolated olfactory receptor neurons from Necturus maculosus.

Olfactory receptor neurons were isolated without enzymes from the mudpuppy, Necturus maculosus, and tested for chemosensitivity. The cells responded to odorants with changes in firing frequency and alterations in excitability that were detected with tight-seal patch electrodes using on-cell and whole-cell recording conditions. Chemosensitive cells exhibited two primary response characteristics: excitation and inhibition. Both types of primary response were observed in different cells stimulated by mixtures of amino acids as well as by the single compound L-alanine, suggesting that there may be more than one transduction pathway for some odorants. Using the normal whole-cell recording method, the chemosensitivity of competent cells washed out rapidly; a resistive whole-cell method was used to record odorant responses under current-clamp conditions. In response to chemical stimulation, excitability appeared to be modulated in several different ways in different cells: odorants induced hyperpolarizing or depolarizing receptor potentials, elicited or inhibited transient, rhythmic generator potentials, and altered excitability without changing the membrane potential or input resistance. These effects suggest that olfactory transduction is mediated through at least three different pathways with effects on four or more components of the membrane conductance. Polychotomous pathways such as these may be important for odor discrimination and for sharpening the "odor image" generated in the olfactory epithelium.

Action Potentials↗

PubMed1991

Cytoplasmic distribution of poly(A)-containing RNA in developing Necturus maculosus oocytes with reference to annulate lamellae.

The cytoplasmic distribution of poly(A)+ mRNA and its relationship to annulate lamellae were examined in developing Necturus maculosus oocytes by in situ hybridization with [3H]poly(U). The specificity of [3H]poly(U) binding was tested by incubating control ovarian sections with either KOH or RNase A before in situ hybridization. In both experiments, the silver grain densities were markedly reduced. Poly(A)+ RNA is uniformly distributed in the cytoplasm until the mid-growth phase and then later in vitellogenesis becomes localized in the subcortical ooplasm. The silver grain density in the cytoplasm varied during oogenesis and was greatest in previtellogenic oocytes. Annulate lamellae commonly are observed with the light microscope in oocytes prior to vitellogenesis. In such oocytes, the labeled mRNA probe is observed over cytoplasmic regions of annulate lamellae. The results suggests that a differential localization of messenger RNA occurs during oogenesis in Necturus maculosus. Furthermore, poly(A)+ RNA is present in cytoplasmic regions of annulate lamellae.

Animals↗

PubMed1983

Acid-base and ionic balance in Ambystoma tigrinum and Necturus maculosus during hypercapnia.

The aquatic urodeles Ambystoma tigrinum and Necturus maculosus responded to hypercapnia quite differently. A. tigrinum, after 2-h exposure to 22 Torr partial pressure of CO2 (PCO2), decreased arterial pH (pHa) from 7.85 to 7.32 and increased arterial pressure of CO2 (PaCO2) to 26 Torr. Plasma [HCO-3] [( HCO-3]pl) remained constant at about 17 mM. Prolonged exposure (24 h) led to a 26% extracellular compensation as pHa rose to 7.46 while [HCO-3]pl increased to 24 mM. Plasma [K+] increased and [Cl-] decreased while [Na+] remained unchanged. Recovery in normocapnic water reversed these changes. N. maculosus did not display similar compensatory changes. Two-hour exposure to 17 Torr PCO2 resulted in a decline of pHa from 7.66 to 7.24, which was not compensated (pHa = 7.19) after 24 h. There were no significant changes in plasma [Na+], [K+], [Cl-], or [HCO-3]. The pHa decline reversed after recovery in normocapnic water, however. The fact that compensation for hypercapnic in A. tigrinum was accompanied by changes in Cl- and K+ concentrations may indicate the participation of epithelial transport mechanisms involving these ions in acid-base balance.

Acid-Base Equilibrium↗

PubMed1979

Morphological studies on the mouth cavity of urodeles. IV. The teeth of the upper jaw and the palate in Necturus maculosus (Rafinesque) (Proteidae: Amphibia).

Adults of the neotenic (paedomorph) Necturus maculosus possess in the upper jaw and the palate rather uniform, conical, monocuspid teeth arranged in a single line ("Zahnzeile"; monostichous pattern) and showing a broad dividing zone, which separates the pedicel and the distal crown. This zone consists of globular and fibrous material obviously mineralized. Both premaxillaries are provided with teeth by one continuous dental lamina, whereas the vomer and palatopterygoid are accompanied by two dental laminae on each side of the palate. The results are compared to the conditions observed in other larval, neotenic, and metamorphosed Urodela.

Animals↗

PubMed1984

Seasonal variations in the fine structure of the Necturus maculosus urinary bladder epithelium: low transporters and high transporters.

Although the urinary bladder of Necturus maculosus provides an important model system for studying the mechanisms of active Na absorption, little critical attention has been paid to the fine structure of its epithelium. Moreover, two distinct groups of urinary bladders, low and high Na transporters, have been described based on short-circuit current or transepithelial potential difference. In the present study, over an 11-month period, stable electrical parameters (short-circuit current, transepithelial potential difference, and resistance) were recorded from 63 chamber-mounted bladders. Analysis of these parameters revealed a highly significant difference between two groups (low transporters and high transporters) occurring at different times of the year. Consistent with these data, in urine collected from the bladders, the Na concentration in low transporters was significantly higher than that in high transporters. A subpopulation of these bladders was subsequently fixed and examined at the light and/or electron microscopic level. Low-transporting bladders were characterized unequivocally by a thin, stratified squamous epithelium only 6-15 micron thick. High-transporting bladders were composed predominantly of columnar-shaped granular cells up to 70 micron in height, with ciliated, mitochondria-rich, and basal cells present in small numbers. There is thus a correlation between transport activity, as measured by electrophysiological techniques and urine sodium analysis, and the structure of the tissue. Moreover, these parameters exhibit significant seasonal variation, the underlying mechanisms of which remain obscure.

Animals↗

PubMed1989

Electron-cytochemical localization of alkaline phosphatase to G cells of Necturus maculosus antrum.

Electron-cytochemical localization of alkaline phosphatase activity was performed on G cells of Necturus maculosus antral mucosa. Alkaline phosphatase activity was localized to the nuclear membrane, the Golgi/endoplasmic reticulum, and the limiting membranes of G cell peptide-secretion vesicles. There was no specific localization of alkaline phosphatase activity to the plasma membrane. Treatment of the tissues with levamisole (an alkaline phosphatase inhibitor) did not markedly reduce the specific alkaline phosphatase activity. Specific lead deposition was reduced by removal of the substrate from the reaction mixture. The results from this study on N. maculosus G cells demonstrate that alkaline phosphatase activity can be found in a non-mammalian gastric endocrine cell and that specific activity was localized primarily to those intracellular structures involved with protein biosynthesis.

Alkaline Phosphatase↗

PubMed1989

Effect of bumetanide on tubuloglomerular feedback in Necturus maculosus.

A non-invasive technique was developed to measure single-nephron glomerular blood flow (SNGBF) in Necturus maculosus. Erythrocytes labelled with rhodamine, a fluorescent dye, were injected systemically and the frequency at which labelled cells entered an arteriole was measured. Frequency was converted to flow by measuring the concentration of labelled erythrocytes in whole blood. Dependence of SNGBF on flow rate in early distal tubules was used to assess tubuloglomerular feedback (TGF). SNGBF decreased with increasing flow in the early distal tubule in a pattern typical of TGF; SNGBF decreased 25% at the highest flow rates. SNGBF increased when bumetanide was added to the perfusate, but the TGF response to flow rate persisted. IC50 (concentration that produces half-maximal inhibition) was 2.4 x 10(-10), 9.8 x 10(-10) and 1.2 x 10(-9) M bumetanide at distal perfusion rates of 5, 10 and 20 nl min-1 respectively. These results are consistent with modulation of SNGBF according to the rate of luminal entry of NaCl into early distal tubule cells. This transport rate depends on the luminal concentration of NaCl, which is tubular flow rate-dependent; NaCl and bumetanide compete.

Animals↗

PubMed1983

Influence of Ca2+ on the mechanosensitivity of the hair cells in the lateral line organs of Necturus maculosus.

The degree of synchronization (DOS) between the afferent spike activity from one stitch of the lateral line of Necturus maculosus (in vivo) and the mechanical stimulation of one neuromast of the same stitch was measured under different circumstances. The DOS was found to be independent of changes in the concentration of monovalent cations (Na+, K+ and choline+) in the bulk solution at high Ca concentration (1 mM). DOS was also independent of the Ca concentration in the range 1 mM-1 microM in Tris-HCl buffer, but was markedly reduced at Ca = 10 microM in MOPS-KOH buffer. The reduced DOS, however, could be restored by addition of 10-20 mM KCl. 5 mM of 4-aminopyridine did not influence the DOS at high Ca concentration, but completely reduced DOS at Ca = 10 microM. D600 (a methoxy derivative of verapamil) decreased DOS both at high and low Ca concentration.

4-Aminopyridine↗

PubMed1984

Electron microscopic study of the innervation of the renal tubules and urinary bladder epithelium in Rana catesbeiana and Necturus maculosus.

The fine structure of the kidney and the bladder of the bullfrog (Rana catesbeiana), the bullfrog tadpole, and the mudpuppy (Necturus maculosus) were studied with special attention to the innervation of renal tubule cells and bladder epithelial cells. In the bullfrog kidney, nerve terminals and varicosities were frequently associated with the tubule cells, apparently in an increasing order from the proximal tubule to the connecting tubule. Although these terminals and varicosities did not directly contact the tubular cell membrane, an aggregation of synaptic vesicles on the side facing the tubule was considered as morphological evidence that neurotransmitter can be released here and can affect the transport activity of the tubule cells. The association of nerve varicosities with canaliculi cells in the connecting tubule was also demonstrated. In the bullfrog tadpoles, renal tubule cells were occasionally innervated. In the mudpuppy, renal tubule cells were only poorly innervated. The epithelium of the bullfrog bladder was commonly innervated. Nerve terminals with synaptic vesicles were located very near basal cells and even contacted them directly on rare occasions. In the mudpuppy, the innervation of the bladder epithelium was observed infrequently. The bullfrog tadpoles did not possess an apparent bladder. In all materials studied, renal arterioles and bladder smooth muscle cells were innervated.

Animals↗

PubMed1994

Cotransport of K+, Cl- and H2O by membrane proteins from choroid plexus epithelium of Necturus maculosus.

1. The interaction between K+, Cl- and H2O fluxes was studied in the ventricular membrane of the choroid plexus epithelium from Necturus maculosus by means of ion-selective microelectrodes. 2. Three experimental strategies were adopted: the osmolarity of the ventricular solution was increased abruptly by addition of (i) mannitol or (ii) KCl; (iii) Na+ in the ventricular solution was replaced isosmotically by K+. 3. The mannitol experiments showed that H2O had two pathways across the ventricular membrane. One was purely passive, with a water permeability, L'p, of 0.64 x 10(-4) cm s-1 (osmol l-1)-1. This operated in parallel with an ion-dependent pathway of similar magnitude which was abolished in Cl(-)-free solutions. 4. When KCl was added there was a flow of H2O into the cell. Surprisingly, this took place despite the osmotic gradient which favoured an efflux of H2O. The effect was blocked by frusemide (furosemide), in which case KCl had the same effects as applications of NaCl or mannitol. 5. Replacement of Na+ with K+ caused an influx of H2O. This flux could proceed against osmotic gradients implemented by mannitol. 6. The present data and those of earlier publications show that the interdependence of the fluxes of K+, Cl- and H2O in the exit membrane can be described as cotransport. The fluxes have a fixed stoichiometry of 1:1:500, the flux of one species is able to energize the flux of the two others, and the transport exhibits saturation and is specific for K+ and Cl-. 7. A molecular model based upon a mobile barrier in a membrane spanning protein gives an accurate quantitative description of the data.

Animals↗