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PubMed1999

Morphological variation of hypaxial musculature in salamanders (Lissamphibia: caudata).

Despite the acknowledged importance of the locomotory and respiratory functions associated with hypaxial musculature in salamanders, variation in gross morphology of this musculature has not been documented or evaluated within a phylogenetic or ecological context. In this study, we characterize and quantify the morphological variation of lateral hypaxial muscles using phylogenetically and ecologically diverse salamander species from eight families: Ambystomatidae (Ambystoma tigrinum), Amphiumidae (Amphiuma tridactylum), Cryptobranchidae (Cryptobranchus alleganiensis), Dicamptodontidae (Dicamptodon sp.), Plethodontidae (Gyrinophilus porphyriticus), Proteidae (Necturus maculosus), Salamandridae (Pachytriton sp.), and Sirenidae (Siren lacertina). For the lateral hypaxial musculature, we document 1) the presence or absence of muscle layers, 2) the muscle fiber angles of layers at mid-trunk, and 3) the relative dorsoventral positions and cross-sectional areas of muscle layers. Combinations of two, three, or four layers are observed. However, all species retain at least two layers with opposing fiber angles. The number of layers and the presence or absence of layers vary within species (Necturus maculosus and Siren lacertina), within genera (e.g., Triturus), and within families. No phylogenetic pattern in the number of layers can be detected with a family-level phylogeny. Fiber angle variation of hypaxial muscles is considerable: fiber angles of the M. obliquus externus range from 20-80 degrees; M. obliquus internus, 14-34 degrees; M. transversus abdominis, 58-80 degrees (acute angles measured relative to the horizontal septum). Hypaxial musculature comprises 17-37% of total trunk cross-sectional area. Aquatic salamanders show relatively larger total cross-sectional hypaxial area than salamanders that are primarily terrestrial.

Anatomy, Cross-Sectional↗
PubMed1986

Secretion of prolactin-synergizing activity (synlactin) by the liver of ectothermic vertebrates in vitro.

Recent work in our laboratory indicated that the liver of rats and pigeons secretes a prolactin synergist (synlactin) in vitro. We have investigated the secretion of this activity by the liver of nine species of ectothermic vertebrates. Liver from three teleosts (goby, salmon, and tilapia), four amphibians (Ambystoma, Necturus, bullfrog, and grass frog) and two reptiles (turtle and anole) was diced, washed, and incubated for 3 hr in isotonic medium. After dialysis, the liver incubation media (LIM) were tested with and without prolactin (PRL) in the local pigeon crop-sac bioassay. The LIM for turtle, larval bullfrog, freshwater salmon, and 5% seawater-adapted goby significantly augmented the local crop-sac response to PRL, but the LIM from anole, adult bullfrog, grass frog, fasted larval Ambystoma, Necturus, 100% seawater-adapted goby, and tilapia did not contain synergizing activity. We conclude that synlactin is secreted by the liver of several species representing three of the major ectothermic classes of vertebrates. It is significant that in two cases, larval bullfrog and 5% seawater-adapted goby, the presence of synlactin occurs in physiological states in which PRL is active. In the opposite cases (adult frog and 100% seawater-adapted goby) the activity was not detectable. We also found that the liver of larval and adult bullfrogs and tilapia released a factor in vitro that had proliferative activity in the crop-sac. This activity appears to be distinct from synlactin.

Amphibians↗
PubMed1977

Length and interspersion of repetitive and non repetitive DNA sequences in four amphibian species with different genome sizes.

The interspersion period of repetitive and unique sequences was analyzed by two different methods, electron microscopy and agarose gel electrophoresis, for four Amphibian species with different nuclear DNA content, namely the Anura Xenopus laevis (3 pg DNA per haploid genome) and Bufo bufo (7 pg) and the Urodela Triturus cristatus (23 pg) and Necturus maculosus (52 pg). Within each of the two subclasses it has been found that interspecific differences, in DNA content, due to variations in the amount of repetitive sequences, do not involve variations in length of the interspersed repetitive sequences. They remain about 380 base pairs. Furthermore, the unique sequences length has been found to be shorter in Bufo (760 base pairs) than in Xenopus (1600) and in Necturus (880) than in Triturus (1340). A study of the interspersion period has shown that the great difference in DNA content between Anura and Urodela, which had been previously shown not to have involved changes in the relative amounts of the various sequence classes, does not involve changes in the interspersion period.

Animals↗
PubMed1975

Electrical properties of amphibian urinary bladder epithelia. I. Inverse relationship between potential difference and resistance in tightly mounted preparations.

In an attempt to find a high-resistance epithelium suitable for microelectrode work, we have studied the electrical properties of Necturus and Amphiuma urinary bladders in comparison to toad bladder. Improved mounting techniques were developed, which yield better reproducible degrees of distension and prevent electrical leaks around the edge of the preparation in the Ussing chamber. Transepithelial potential difference and resistance was measured with NaCl Ringer's on either surface of the epithelium, as well as under conditions of ion substitutions and in the presence of amiloride. Compared to data from conventionally mounted toad bladders reported in the literature, our experiments yielded higher potential differences and resistances in all three species. In Necturus values of up to 175 mV and 75 komega cm2 were recorded. Furthermore an inverse relationship was observed between potential difference and resistance, which was not noticed previously with the conventional mounting technique. The relationship is discussed quantitatively in terms of the two-membrane model of active Na+ transport, for which it provides further supportive evidence.

Amiloride↗
PubMed1975

The electrical potential profile of gallbladder epithelium.

In this study the relative ionic permeabilities of the cell membranes of Necturus gallbladder epithelium have been determined by means of simultaneous measurement of transmural and transmucosal membrane potential differences (PD) and by ionic substitution experiments with sodium, potassium and chloride ions. It is shown that the mucosal membrane is permeable to sodium and to potassium ions. The baso-lateral membrane PD is only sensitive to potassium ions. In both membranes chloride conductance is negligible or absent. The ratio of the resistances of the mucosal and baso-lateral membranes, RM/RS, increases upon reducing the sodium concentration in the mucosal solution. The same ratio decreases when sodium is replaced by potassium which implies a greater potassium than sodium conductance in the mucosal membrane. The relative permeability of the shunt for potassium, sodium and chloride ions is: PK/PNa/PCl=1.81:1.00:0.32. From the results obtained in this study a value for the PK/PNa ratio of the mucosal membrane could be evaluated. This ratio is 2.7. From the same data the magnitude of the electromotive forces generated across the cell membranes could be calculated. The EMF's are -15mV across the mucosal membrane and -81mV across the baso-lateral one. Due to the presence of the low resistance shunt the transmucosal membrane PD is -53.2mV (cell inside negative) and the transmural PD is +2.6mV (serosal side positive). The change in potential profile brought about by the low resistance shunt favors passive entry of Na ions into the cell across the mucosal membrane. Calculations show that this passive Na influx is maximally 64% of the net Na flux estimated from fluid transport measurements. The C-1 conductive of the baso-lateral membrane is too small to allow electrogenic coupling of C1 with Na transport across this membrane. Experiments with rabbit gallbladder epithelium indicate that the membrane properties in this tissue are qualitatively similar to those of Necturus gallbladder epithelium.

Amiloride↗
PubMedDate not supplied

Basolateral KCl co-transport in a NaCl-absorbing epithelium.

In NaCl-absorbing epithelia such as proximal renal tubule, small intestine and gallbladder, Na+-dependent Cl- entry across the luminal membrane is an electroneutral transport process that has been attributed to Na-Cl symport, Na-K-Cl symport, or a double (Na-H, Cl-HCO3) antiport. At the basolateral (antiluminal) membrane, Na+ extrusion can be attributed to the Na+-K+ pump, and Cl- transport could be explained in principle by electrodiffusion since the intracellular Cl- activity is higher than predicted for equilibrium distribution. However, in Necturus gallbladder, as in other epithelia, the electrodiffusional Cl- permeability of the membrane (PCl) is too low to account for the transepithelial Cl- transport rate. Because K+ is at a higher chemical potential in the cell than in the extracellular fluid, and because serosal Cl- substitutions have only small effects on membrane potential, the hypothesis of carrier-mediated electroneutral KCl co-transport was proposed. The experiments reported here were designed to test this hypothesis in Necturus gallbladder epithelium. Intracellular Cl- and K+ activities (aCli, aKi) were measured with ion-sensitive intracellular microelectrodes before, during and after ionic substitutions of the serosal (basolateral) bathing medium. The results demonstrate a Na+-independent basolateral membrane KCl symport.

Biological Transport, Active↗
PubMed1977

Effects of acetylsalicylate on alkalinization, acid secretion and electrogenic properties in the isolated gastric mucosa.

The effects of acetylsalicylate (ASA) on the in vitro secretory and electrical properties of Necturus and Rana temporaria gastric mucosa have been studied. The gastric antrum alkalinized the luminal surface, while in the fundus it is likely that acidificaiton and alkalinization occur simultaneously and that net secretion is due to the dominance of one or other of these processes. The histamine H2 receptor antagonist Metiamide was used to inhibit acid secretion for studies on fundic alkalinization in Rana temporaria. Submucosal application of 3 mM ASA for 30 min markedly reduced alkalinization in the antrum and the frog fundus. Following removal of ASA there was only partial recovery of this secretion. The drug caused slight inhibtion of spontaneous acid secretion in Necturus fundus but not of histamine-stimulated acid secretion in the frog fundus. Following salicylate removal, the rate of acid secretion increased to a higher level than before administration in both tissues. There was a small flux of ASA across the mucosa which was greatest in the acid secreting frog fundus (4.38mumol cm-2 min-1) and least in the antrum (2.19 mumol cm-2 min-1). Esposure of the gastric mucosa to ASA was generally associated with a fall in transmucosal electric potential difference and short-circuit current together with an increase in electrical resistance. It is proposed that the greater sensitivity of alkalinization to ASA is responsible for the apparent increase in the rate of acid secretion which occured.

Animals↗
PubMed1970

The contribution by glial cells to surface recordings from the optic nerve of an amphibian.

1. The contribution by glial cells to surface recordings has been examined in the optic nerve of the amphibian Necturus maculosus. The method of current injection was employed selectively to alter the membrane potential of glial cells without affecting that of the axons. The resulting changes in potential were recorded simultaneously from the surface of the nerve using the sucrose gap method and intracellularly from a glial cell near the gap.2. The sucrose gap method recorded 40% of the changes in glial membrane potential. This percentage was not affected when the current electrode was inserted into different glial cells while maintaining the recording conditions constant.3. Following axonal degeneration, produced by removing the eye 2-3 months earlier, the percentage contribution by glia increased to 84%.4. By measuring sucrose gap responses to changes in K(o) it was possible to estimate that the sucrose gap method recorded 31-60% of changes in axonal membrane potential. It was also determined that the axons, unlike glial cells, are relatively insensitive to reductions in K(o). Surface responses to decreases in external potassium thus reflect the magnitude of the glial contribution.5. It is concluded that changes in glial membrane potential contribute about as much to surface recordings from the optic nerve of Necturus as do equivalent changes in axonal membrane potential. The contributions by the glial cells and axons are related to the relative volumes of tissue they respectively occupy. The significance of these findings to the analysis of surface recordings from the mammalian brain is discussed. Since mammalian glial cells, like those in Amphibia and the leech, become depolarized during neuronal activity and on the basis of electron microscopic evidence appear to be electrically coupled, it is likely that they contribute to the electroencephalogram.

Animals↗
PubMed1975

Ion transport by amphibian antrum in vitro. I. General characteristics.

Both Necturus and bullfrog antrum show stable PD, resistance, and short-circuit current (Isc) when mounted in an Ussing chamber. Measurements of Na+ and Cl minus flux showed that both ions are actively transported across Necturus antrum, Na+ from secretory to nutrient, Cl minus from nutrient to secretory (both net fluxes being similar to 0.30 mueq cm minus 2 h minus 1). Only the Na+ transport contributed to the Isc and PD as evidenced by a) Na+ removal, b) the effects of amiloride on the secretory surface, c) the effects of ouabain on the nutrient side. Microelectrode experiments confirm the Na+ conductance of the secretory cell membrance, a HCO3 minus conductance of both cell membranes, and a KCl conductance across the nutrient cell membrane. In addition, antrum apparently secretes alkali (similar to 0.35 mueq cm minus 2 h minus 1), which secretion is sensitive to metabolic inhibitors and Diamox. Nutrientside HCO3 minus increased the rate of alkaline secretion and a transmucosal HCO3 minus gradient could contribute to ISC and PD. A model is proposed to account for the electrical properties of the tissue.

Amiloride↗
PubMed1994

Julia B. Platt (1857-1935): pioneer comparative embryologist and neuroscientist.

Julia Barlow Platt was a comparative embryologist and neurobiologist who was primarily interested in segmentation of the head in vertebrates. She was born on September 14, 1857 in San Francisco, California. Platt grew up in Burlington, Vermont, attended the University of Vermont and began graduate studies at Harvard University. Her nine years as a graduate student were spent on two continents with some of the most influential comparative zoologists of the time. Platt's remarkable scientific accomplishments over a ten year period include a description of axial segmentation currently used in the staging of chick embryos and the first description of a separate anterior head segment in Squalus embryos. Her most controversial study identified ectodermal cells in Necturus embryos that gave rise to head cartilage and dentine, a discovery which was the impetus for the reassessment and modification of the germ layer concept. She was one of the first women to 'matriculate' at a German university and receive a Ph.D. degree. Platt played a pioneer role in opening opportunities for other women who followed her. Platt was one of the first women neuroscientists. Among her contributions, she distinguished dorsolateral placodes, epibranchial placodes, and the first stages of lateral line organs in Necturus, and she described nerve fibers originating in the spinal cord and extending to the notochord in Branchiostoma (= Amphioxus). After receiving a Ph.D. degree in Freiburg, Germany in 1898, Platt was unable to secure a suitable teaching position and, as a result, her scientific career came to an end. She retired to Pacific Grove, California, where she pursued civic duty with the same vigor and energy she had dedicated to scientific research. We provide a sketch of her remarkable life and work as a comparative embryologist, neuroscientist and civic leader.

Embryology↗
PubMed1974

Pressure control of sodium reabsorption and intercellular backflux across proximal kidney tubule.

The magnitude of changes in luminal hydrostatic pressure (DeltaP(L)), peritubular capillary hydrostatic pressure (DeltaP(PT)), and peritubular capillary colloid osmotic pressure (Deltapi) was determined in the Necturus kidney during volume expansion (VE). The specific effects of separate changes of each pressure parameter on proximal net sodium transport (J(Na)) were studied in isolated perfused kidneys. The combined effect of DeltaP(L), DeltaP(PT), and Deltapi, of a magnitude similar to that induced by volume expansion, decreases J(Na) by 26% in the perfused kidney. A major portion of the natriuresis in VE is due to changes in intrarenal pressures. The effect of Deltapi on the permeability characteristics of Necturus proximal tubule was studied. With increasing Deltapi, the ionic conductance of the paracellular shunt pathway decreased, since transepithelial input and specific resistance rose significantly, whereas cellular membrane resistance remained unchanged. Transepithelial permeability coefficients for sodium chloride and raffinose changed inversely proportional to transepithelial resistance, indicating an alteration of a paracellular permeation route. Net passive sodium backflux and active transport flux components were calculated. Increased net sodium transport with rising Deltapi is accompanied by a significant drop in passive back diffusion, without an increment in the active flux component. Change in passive sodium ion back diffusion thus appears to be a key physiological factor in the control of transepithelial sodium transport.

Animals↗
PubMed1977

Properties of gastric antrum. III. Selectivity and modification of shunt conductance.

The permselectivity of the paracellular pathway of amphibian (Necturus and bullfrog) antrum was investigated with respect to intracationic selectivity and the K+ and Cl- permeability ratio as a function of mucosal pH. The intracationic selectivity sequence was Rb+ greater than K+ greater than Cs+ greater than Na+ greater than Li+. Both antra showed weak cationic selectivity at pH 7.4, and at pH 4.4 for bullfrog and pH 3.0 for Necturus, the ratio Pk+/P Cl- was unity. At lower mucosal pH the tissues were anion selective. Treatment of the tissue with a water-soluble carbodiimide enhanced anion selectivity at higher pH; carbenoxolone, a weak acid, resulted in maintained cation selectivity at lower pH. These data suggest that carboxyl groups play a role in determining shunt selectivity; the increase in anion selectivity below pH 2.0 suggests that phosphate or sulfate groups could also be involved.

Amphibians↗
PubMed1986

Catecholamine, serotonin, and substance P-like peptide containing intrinsic neurons in the mudpuppy parasympathetic cardiac ganglion.

The mudpuppy cardiac ganglion contains 2 neuron types: large parasympathetic postganglionic projection neurons and smaller intrinsic neurons originally described by McMahan and Purves (1976) as intensely fluorescent (SIF) cells. The function of these SIF cells, present in the mudpuppy cardiac ganglion, is unknown. Further, direct application of catecholamines, which are thought to be contained in SIF cells, to the parasympathetic postganglionic cells has no effect (Hartzell et al., 1977). As SIF cells in other ganglion preparations recently have been shown to contain putative transmitter substances in addition to catecholamines, immunocytochemical experiments were conducted to test for the presence of additional transmitter substances in the SIF cells within the cardiac ganglion. Whole-mount septal preparations were dissected from Necturus maculosus and processed for indirect immunocytochemistry. The results indicated that many of these intrinsic neurons contained 5-HT or a substance P-like peptide, or both. Many small intrinsic neurons which contain either substance P or 5-HT were also positive for aqueous-aldehyde-induced fluorescence, indicating the presence of a catecholamine. Finally, some of these cells appeared to contain all 3: a catecholamine, 5-HT, and a substance P-like peptide.

Animals↗
PubMed1996

Development of voltage-dependent currents in taste receptor cells.

Taste buds, the specialized end organs of gustation, comprise a renewing sensory epithelium. Undifferentiated basal cells become taste receptor cells by elongating and extending processes apically toward the taste pore. Mature taste cells are electrically excitable and express voltage-dependent Na+ Ca2+, and K+ currents, whereas basal stem cells exhibit only slowly activating K+ currents. The question we have addressed in the present study is whether contact with the taste pore is required for expression of voltage-dependent inward currents in Necturus taste cells. Mature taste cells were distinguished from developing cells by labeling the apical surface of the cells with fluorescein-isothiocyanate-conjugated wheat germ agglutinin (FITC-WGA), while the tissue was still intact. Elongate cells lacking FITC-WGA staining were interpreted as developing taste cells that had not yet reached the taste pore. Giga-seal whole-cell recording revealed that most developing taste cells lacked inward currents. Although some developing cells expressed inward currents, they were much smaller than those of mature cells, and the activation kinetics of the K+ currents were slower than in mature cells. Electron microscopy confirmed the identity of labeled and unlabeled cells. All FITC-WGA-labeled cells exhibited the ultrastructural characteristics of mature taste receptor cells, whereas most unlabeled taste cells had a characteristic morphology that was markedly different from mature taste receptor cells or basal stem cells. These data suggest that contact with the taste pore is required for the development of inward currents in taste cells.

Animals↗
PubMed1997

Pump-leak parallelism in sodium-absorbing epithelia: the role of ATP-regulated potassium channels.

In all Na(+)-absorbing and Cl(-)-secreting epithelia, an increase in the activity of the Na+,K(+)-pump at the basolateral membrane is accompanied by an increase in the K+ conductance of that barrier and vice versa. We have recently identified an ATP-regulated K+ channel, K(ATP), in basolateral membrane vesicles isolated from Necturus maculosa small intestinal epithelial cells that could be responsible for this parallelism between pump activity and leak. Thus, an increase in pump activity would result in a decrease in local ATP activity and an increase in local ADP activity and, in turn, an increase in the open-probability of the channel whereas a decrease in in pump activity would have the opposite effect. Further, the likelihood that the number of pumps far exceeds the number of leaks per unit area of membrane suggests that the ATP and ADP activities that influence K(ATP) channel activity may differ markedly from the "bulk" cytoplasmic values.

Adenosine Diphosphate↗
PubMed1993

Role of apical ion channels in sour taste transduction.

Sour taste perception depends primarily on the concentration of H+ in the taste stimulus. Acid stimuli elicit concentration-dependent action potentials in taste cells. Recent patch-clamp studies suggest that protons depolarize taste cells by direct interaction with apically located ion channels. In Necturus maculosus, the voltage-dependent K+ conductance is restricted to the apical membrane of taste cells. The current flows through a variety of K+ channels with unitary conductances ranging from 30 to 175 pS, all of which are blocked directly by citric acid applied to outside-out or perfused cell-attached patches. In contrast, hamster fungiform taste cells appear to utilize the amiloride-sensitive Na+ channel for acid transduction. Amiloride completely inhibits H+ currents elicited by acid stimuli in isolated taste cells, with an inhibition constant similar to that for amiloride-sensitive Na+ currents (Ki = 0.2 microM). Treatment of isolated taste cells with the bioactive peptide arginine-vasopressin results in similar increases in both the amiloride-sensitive Na+ and H+ currents; the effect is mimicked by 8-bromocyclic AMP. These results suggest that H+ can permeate amiloride-sensitive Na+ channels in hamster fungiform taste cells, contributing to the transduction of sour stimuli.

Animals↗
PubMed1990

Marginal neurons in the urodele spinal cord and the associated denticulate ligaments.

Marginal neurons have been described in the spinal cords of a variety of vertebrates including lamprey, reptiles, birds, and mammals but not in amphibians. There has been speculation about a motor function for these neurons but recent experimental evidence in lampreys indicates that they are intraspinal mechanoreceptor neurons. Additional evidence on reptiles and birds demonstrates that the marginal neurons are closely associated with the denticulate ligaments. In the present investigation, we have examined the spinal cords of Necturus, Ambystoma tigrinum, and A. mexicanum with light and electron microscopic techniques. Marginal nuclei were found in the ventrolateral position immediately internal to the pia and to the denticulate ligament. The marginal neurons were scattered in a continuous column of neuropil without segmental accumulation. They were approximately 30 to 50 microns in diameter and fusiform with dendrites extending from the poles, parallel with the length of the spinal cord. Neuronal fingerlike processes, like those found in peripheral mechanoreceptors and in the marginal nuclei of reptiles, were also found in the three species of urodeles studied. The structure of the denticulate ligaments, similar in the three different amphibians, was composed of collagen, elastin, and fibroblasts, all of which were concentrated in the segmental lateral processes.

Ambystoma↗
PubMed1994

Electrophysiological and morphological properties of light and dark cells isolated from mudpuppy taste buds.

Isolated Necturus taste receptor cells were studied by giga-seal whole-cell recording and electron microscopy to correlate electrophysiological properties with taste cell structural features. Dark (type I) cells were identified by the presence of dense granular packets in the supranuclear and apical regions of the cytoplasm. In response to a series of depolarizing voltage commands from a holding potential of -80 mV, these cells exhibited a transient, TTX-sensitive inward Na+ current, a sustained outward K+ current, and a slowly inactivating inward Ca++ current. Light (type II) cells were identified by a lack of granular packets and by an abundance of smooth endoplasmic reticulum distributed throughout the cell. In addition, isolated light cells had clear vesicular inclusions in the cytoplasm and blebs on the plasma membrane. Light cells were divided into two functional populations based upon electrophysiological criteria: cells with inward and outward currents, and cells with outward currents only. Light cells with inward and outward currents had voltage-activated Na+, K+, and Ca++ currents with properties similar to those of dark cells. In contrast, the second group of light cells had only voltage-activated outward K+ currents in response to depolarizing voltage commands. These data suggest that dark cells and light cells with inward and outward currents are capable of generating action potentials and releasing neurotransmitters onto gustatory afferent neurons in response to taste stimulation. In contrast, light cells with outward currents only likely serve a different function in the taste bud.

Animals↗