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PubMed1977

Sex differences in Necturus urinary bladders.

We describe morphological sex differences in urinary bladders of the urodele amphibian Necturus maculosus. The mucosal epithelial cells of bladders from males were tall and contained considerable PAS-positive material. In scanning electron micrographs of the mucosal surface, epithelial cells from male bladders were well demarcated and were capped with microplicae or with long cilia. The mucosal epithelial cells of bladders from females were low and contained only a small amount of PAS-positive material; in scanning electron micrographs cell boundaries could not be distinguished and no cilia or microplicae were present. Bladders from males had higher transepithelial potential difference and lower water content than bladders from females. Urine analyses were not significantly different in the two sexes. It is suggested that response to androgens in the male accounts for the observed differences.

Animals↗

PubMed1992

Histochemical study of the heart of the axolotl (Ambystoma mexicanum).

We have investigated the presence of cells containing monoamines, substance P, and neuron-specific enolase (NSE) in the heart and in the pericardial wall of a urodele amphibian, the axolotl. Fibers containing substance P-like immunoreactivity were present in the heart but not in the pericardial wall. Also present in the heart were small branched cells, which stained metachromatically with toluidine blue. Similar cells were found in the peritoneum and were tentatively identified as mast cells. NSE-immunoreactive fibers were found both in the heart and in the pericardial wall. Small intensely fluorescent (SIF) cells of the pericardial wall contained a high concentration of norepinephrine but no other monoamines, substance P, or NSE. Comparison with data available for the mudpuppy, Necturus maculosus, a closely related amphibian species, suggests that the innervation of the heart in the axolotl is substantially different.

Ambystoma mexicanum↗

PubMed1975

A further study of the fine structure and membrane properties of neuroglia in the optic nerve of Necturus.

The optic nerve of Necturus maculosus consists of a homogeneous population of astroglia and bundles of unmyelinated axons. The glial cell processes ramify within the nerve roughly delineating fascicles of axons and come together at the periphery to form a complete external limiting membrane interrupted only by narrow clefts between adjacent processes. They are frequently "attached" to one another, forming specialized junctions. Blood vessels are entirely outside the nerve which is surrounded by a basal lamina. The temperature dependence of the glial membrane potential is accurately predicted by the Nernst relation. The membrane potential is unaffected by changes in Cl, Na, Li, and guanidinium which are apparently impermeant. The permeability of the glial membrane to other cations is in the sequence Tl greater than K greater than Rb greater than Cs greater than NH4. This suggests that the chemical nature of the site of potassium permeability in glial cells is similar to that in the neuron.

Animals↗

PubMed1988

Putative neurotransmitters in the retinae of three urodele species (Triturus alpestris, Salamandra salamandra, Pleurodeles waltli).

The immunocytochemical localization of several substances with putative neurotransmitter or modulator properties was investigated in the retinae of three urodele species. Gamma-aminobutyric acid-like immunoreactive labelling appeared in different types of amacrine and horizontal cells. In addition, labelled fibres in the optic nerve were detected. It was not possible to determine whether these fibres were ganglion-cell axons or part of an efferent projection. Endogenous serotonin was found in several populations of amacrine cells including stratified and diffuse types. Glucagon-like immunoreactivity appeared in one bistratified amacrine cell type, and neurotensin-like immunoreactivity was detected in a single monostratified amacrine cell type. Metenkephalin-like-immunoreactive labelling was type. Metenkephalin-like-immunoreactive labelling was rare but found in several sublaminae of the inner plexiform layer. Thus each peptide-like-immunoreactive cell type makes up a distinct and unique population of cells and probably has a special functional role in retinal processing. There are striking similarities in the peptide-like immunoreactive patterns of Triturus alpestris and Necturus maculosus whereas in Ambystomatidae the peptide-like-immunoreactive systems appear to be differently organized. This supports the hypothesis that Salamandridae and Proteidae are more closely related to each other than to the Ambystomatidae.

Animals↗

PubMed1976

DNA reassociation kinetics in relation to genome size in four amphibian species.

DNA reassociation kinetics were studied, by means of the hydroxyapatite chromatography method, for four species of Amphibians with different nuclear DNA content: Xenopus laevis (3 pg DNA per haploid genome) and Bufo bufo (7 pg) of the Anura subclass and Triturus cristatus (23 pg) and Necturus maculosus (52 pg) of the Urodela subclass. Within each subclass the two species studied were found to have about the same absolute amount of unique DNA. The differences of total nuclear DNA can be accounted for by quantitative variations of the repetitive sequence classes, at least in part due to changes in the number of copies of the various sequences. On the contrary the great difference in nuclear DNA between the two subclasses, Anura and Urodela, involves all sequence classes in parallel; the slowly reassociating fraction appears to be unique in spite of a tenfold difference in absolute amount. The dependence of reassociation kinetics on DNA fragment length for the four species indicates for all of them an interspersed organization of the various sequence classes.

Amphibians↗

PubMed1982

Transcription of complementary repeat sequences in amphibian oocytes.

Repeat sequences are transcribed in the germinal vesicles of amphibian oocytes. In the hnRNA population both complements of the repeats are found and can be readily detected because they form intermolecular duplex structures. The structure and formation of duplex regions have been studied in the hnRNA of Xenopus laevis, Triturus cristatus, Amphiuma means and Necturus maculosus, a series of amphibians of increasing genome size (C-value). In T. cristatus, the duplex structures are mostly 600-1200 bp in length, whereas in X. laevis they are shorter and in N. maculosus they tend to be longer. Although the proportion of RNA sequence capable of rapidly forming duplex structures is different in different organisms, this property bears no relationship to C-value. However the sequence complexity of complementary repeats, as estimated from the rate of duplex formation, does show an increasing trend with C-value. The complementary repeats found in oocyte hnRNA are transcribed from families of DNA sequence that are each represented in the genome by thousands of copies. The extent of cross-species hybridization is low, indicating that the repeat sequences transcribed in different amphibian genera are not the same. In situ hybridization experiments indicate that the repeat sequences are spread throughout the genome. The evolution and possible function of complementary repeats are considered.

Amphibians↗

PubMed1975

[The role of cyclic adenosine 3':5'-monophosphate as "second messenger" in gastric acids secretion (author's transl)].

The criteria to determine whether cyclic adenosine 3':5'-monophosphate (cyclic AMP) is or is not involved in a particular hormone response require to look for positive qualitive, quantitative and temporal correlations between the effects of the hormone on cyclic AMP levels and the physiological response. These requirements have been shown to be fulfilled in histamine and pentagastrin-stimulated gastric acid secretion of the frog (Necturus maculosus) and the rat. In dog and man, however, the available evidence does not support a role for cyclic AMP in the gastric secretory process; it remains a challenge for future research whether in those species cyclic guanosine 3':5'-monophosphate acts as an intracellular substitute for cyclic AMP.

Adenylyl Cyclases↗

PubMed1987

The effects of chloride ions on electrodiffusion in the membrane of a leaky epithelium. Studies of intact tissue by microelectrodes.

The electrodiffusive permeability for Cl-, its dependence on low extracellular Cl--concentrations and the interaction between the movements of Cl- and K+ were investigated in the ventricular membrane of epithelial cells from the choroid plexus of Necturus maculosus. Cells were probed with ion-selective microelectrodes sensitive to Cl-, K+ and H+. The initial effects of abrupt changes in the Cl--concentration (Cl-v) and/or the K+-concentration (K+v) of the ventricular solution were investigated. The effect of changing the membrane potential by changing K+v was twofold: It caused an electrodiffusive flux of Cl- via a permeability of 1.3 X 10(-6) cm s-1. This permeability together with the K+-permeability of the ventricular membrane (24 X 10(-6) cm s-1) determined the membrane potential in the given steady state within a few mV. The other effect of the depolarization was an increase in the intracellular concentration of HCO-3 which in turn caused an influx of Cl- via electroneutral Cl-/HCO-3 exchange. The Cl--permeability was reduced by more than 60% and the neutral exchange by more than 90% by furosemide. The effect of decreases in Clv was a tenfold increase of the electrodiffusive Cl--permeability of the ventricular membrane to 12.2 X 10(-6) cm s-1 and also a tenfold increase in the permeability to K+. This activation was reduced by two thirds by furosemide, and by depolarizations of the cell by high K+v. In the given steady state the HCO-3/Cl- exchanger at the ventricular membrane transports at a rate of 300 pmol cm-2 s-1 and moves Cl- into the cell and HCO-3 into the ventricular solution. Thus the epithelium alkalinizes the cerebrospinal fluid at a rate which is about three times faster than the net transport rate of Na+.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1978

Clustered and interspersed repetitive DNA sequences in four amphibian species with different genome size.

We have compared the amount of clustered and interspersed repetitive sequences in the genome of four Amphibia with different DNA contents per haploid nucleus: two Anura (Xenopus laevis, 3 pg and Bufo bufo, 7 pg) and two Urodela (Triturus cristatus, 23 pg and Necturus maculosus, 52 pg). High molecular weight DNA of the four species was denatured and reassociated to the same Cot in order to obtain duplex sequences with a similar reiteration frequency. Single-stranded DNA was digested off with the Aspergillus S1 nuclease. DNA was then fractionated according to the molecular weight through an agarose A-50 column. We found that the amount of long repetitive sequences is roughly proportional to the genome size in the four species, while the number of short (about 300 base pairs) repetitive sequences is increased many-fold in the species with the larger DNA content, both in Anura and in Urodela.

Amphibians↗

PubMed1979

Pathways and polarities of synaptic interactions in the inner retina of the mudpuppy: I. Synaptic blocking studies.

Intracellular recording techniques were applied to the perfused retina-eyecup of the mudpuppy (Necturus maculosus). The use of input resistance measurements and the application of cobalt as a synaptic blocking agent provided evidence about the mode of synaptic operation for depolarizing and hyperpolarizing bipolar cell input to cells of the inner retina. In addition, cobalt produced a more rapid block of hyperpolarizing bipolars compared to depolarizing bipolars; in some postbipolar neurons, a temporary simplification of neuronal response waveform was observed and such cells resembled the depolarizing bipolar. These experiments suggest that bipolar cell input is excitatory and that response characteristics of third-order neurons is partially dependent on the relative input from the two types of bipolar cells.

Animals↗

PubMed1979

Pathways and polarities of synaptic interactions in the inner retina of the mudpuppy: II. Insight revealed by an analysis of latency and threshold.

Intracellular recording experiments in the mudpuppy (Necturus maculosus) have demonstrated that depolarizing bipolars show a one-half to one log unit higher threshold to light stimuli and a longer latency when compared to hyperpolarizing bipolars. This threshold difference cannot be ascribed to differences in rod and cone connections. Experimentally it was possible to use these differences to evaluate postbipolar cell connections; such experiments support the idea that some neurons are connected to one or the other bipolar cell type, while on-off cells receive input from both.

Animals↗

PubMed1990

Neurotensin is localized to paracrine cells in the urinary bladder of the turtle, Chrysemys picta.

Urinary bladder from the painted turtle, Chrysemys picta, contains a substantial population of endocrine/paracrine cells scattered through the mucosal epithelium which immunostains using antisera directed toward mammalian neurotensin (NT). Radiommunoassay of 0.1 N HCl extracts of Chrysemys bladder indicated an immunoreactive NT (iNT) concentration of 161 +/- 39 pmol/g tissue (n = 9), an amount lower but comparable in magnitude to that found in mucosal scrapings of the intestine (926 +/- 125 pmol/g, n = 9). Bladder and intestinal iNT were indistinguishable during chromatography on Sephadex G-25 and HPLC on mu-Bondapak C18 where they eluted at the same position as avian NT. Similar results were obtained by immunocytochemistry and radioimmunoassay of urinary bladders from Pseudemys scripta scripta, P. scripta elegans, and P. floridana, three emydid species closely related to C. picta, but not from Sternotherus odoratus and Trionyx spinifer asper, representatives of the families Kinosternidae and Trionychidae, respectively, although positive results were obtained with intestinal preparations from these species. NT cells were not seen in urinary bladder from Rana pipiens, Bufo marinus, Necturus maculosus, or Anolis carolinensis. Thus, the presence of NT-containing cells in urinary bladder is not common among subavian vertebrates and may even be restricted to the Emydidae family of chelonians.

Animals↗

PubMed2003

Myoseptal architecture of sarcopterygian fishes and salamanders with special reference to Ambystoma mexicanum.

During axial undulatory swimming in fishes and salamanders muscular forces are transmitted to the vertebral axis and to the tail. One of the major components of force transmission is the myoseptal system. The structure of this system is well known in actinopterygian fishes, but has never been addressed in sarcopterygian fishes or salamanders. In this study we describe the spatial arrangement and collagen fiber architecture of myosepta in Latimeria, two dipnoans, and three salamanders in order to gain insight into function and evolution of the myoseptal system in these groups. Salamander myosepta lack prominent cones, and consist of homogenously distributed collagen fibers of various orientations that never form distinct tendons. Fiber orientations are difficult to homologize with those of fish myosepta. The myosepta of Latimeria and dipnoans (Protopterus and Neoceratodus) illustrate that major changes in architecture occurred in the sarcopterygian clade (loss of horizontal septum), in the rhipidistian (dipnoans + tetrapods) clade (loss of epineural and epipleural tendon), and in tetrapods (loss of lateral tendons and myoseptal folding). When compared to fishes, the myosepta of wholly aquatic salamanders (Ambystoma mexicanum, Amphiuma tridactylum, Necturus maculosus) do not have the lateral tendons we suppose serve to transfer muscular forces posteriorly. We propose that alternative structures (most conspicuously present in Ambystoma) perform this function: posteriorly the relative amount of connective tissue increases considerably, and myosepta are disintegrated to horizontal lamellae of connective tissue. The structures thought to be involved in modulation of body stiffness in fishes during swimming are also absent in salamanders. Our data also have implications for the hypothesis that salamander hypaxial myosepta are designed to increase shortening amplification of the hypaxial muscle fibers. The posterior hypaxial myosepta of all three salamander species possess only mediolaterally directed collagen fibers, which would indeed amplify the shortening of the associated muscle.

Journal Article↗

PubMed1972

Calcium dependence of ciliary activity in the oviduct of the salamander Necturus.

1. Ciliary activity in the oviduct of the mud puppy Necturus maculosus was monitored by a photometric technique in normal, decalcified, and Triton X-extracted preparations to investigate the regulatory role of calcium ions.2. The frequency of ciliary beating in the isolated tissue ranged from 0 to 12 beats/sec. The frequency in any one group of cells underwent large cyclical variations with periods of 2 min or more.3. Frequency of beating reached a maximum and remained at a plateau upon addition of 1 mM caffeine. Beating temporarily ceased upon removal of the caffeine.4. Ionophoretic injection of calcium ions into an active cell bathed in Ringer solution produced an increased rate of beating. In cells rendered quiescent by prior decalcification with EGTA, injected calcium rapidly restored ciliary activity.5. Epithelia extracted in Triton X-100 were inactive until reactivated by addition of ATP and magnesium ions. The frequency of beating increased between 0.1 and 4.0 mM ATP in 1 mM magnesium, and between 0.2 and 2.5 mM magnesium in 1 mM ATP.6. The frequency of beating in the ATP-reactivated, Triton-extracted tissue was independent of the calcium ion concentration.7. Cells inactivated by decalcification were reactivated by injection of ATP or by extracellular ATP levels as low as 3 x 10(-8)M.8. It is concluded that the frequency of beating depends directly on the concentration of the available energy source, presumably ATP, and that an indirect dependence of beating frequency on calcium concentration in the living tissue results from rate-limiting effects of intracellular calcium on metabolic steps in pathways leading to ATP synthesis.

Adenosine Triphosphate↗

PubMed1975

Regenerative hyperpolarization in rods.

1. The electrical properties of the rods in Necturus maculosus were studied at the cell body and the outer segments in dark and light under current and voltage clamp with a pair of intracellular electrodes separated by about 1 mum. 2. The membrane resistance in the dark was voltage- and time-dependent both for the cell body and the outer segment. Slight depolarizations in the cell body reduced the slope resistance from 60 to 10 M omega with a time constant of about 1 sec. Polarization in either direction, at the outer segment, when greater than about 20 mV, reduced the slope resistance from 60 to 30 M omega. The dark potential in the cell body was typically -30 to -35 m V; at the outer segment it was typically only -10 to -15 mV. 3. The light-elicited voltage response in both the cell body and the outer segment was largest with the membrane near the dark potential level. In both regions, the response was reduced when the membrane was polarized in either direction. 4. Under voltage-clamp conditions, a reversal potential for the light response near + 10 mV was measured at the outer segment. At the cell body no reversal potential for the light response was measured; there the clamping current required during the light response was almost of the same magnitude at all potential levels. 5. When the membrane at the cell body was hyperpolarized in the dark under voltage clamp, a transient outward current, typically about one-half the magnitude of the initial inward clamping current was required to maintain the membrane at the clamped potential level. This outward current transient was associated with a decrease in membrane resistance with similar time course. The transient outward current reversed and became inward when the membrane was clamped to potentials more negative than -80 mV. Thus, the transient outward current appears to involve a transient activation initiated by hyperpolarization. I is regenerative in that it is initiated by hyperpolarization and tends to further hyperpolarize the membrane. 6. The reversal potential for the light response was measured at the outer segment but not at the cell body. The regenerative hyperpolarization was measured at the cell body but not at the outer segment. Thus, the outer segment and cell body appear to have different electrical properties: a light-elicited resistance increase at the outer segment causes a potential-dependent transient decrease at the inner rod. 7. An electrical model of the rod, based upon estimates of the membrane resistances and membrane e.m.f.s. in the dark, was derived from the data. This model predicts the appropriate response potentials at outer segment and cell body when perturbed by the measured light-elicited resistance increase at the outer segment. An estimate of membrane current in dark, of 0-2 mA, is also derived from the model.

Animals↗

PubMed1977

Synaptic excitation and inhibition resulting from direct action of acetylcholine on two types of chemoreceptors on individual amphibian parasympathetic neurones.

1. Synaptic transmission was studied in visually identified parasympathetic ganglion cells that modulate the heart beat of the mudpuppy Necturus maculosus).2. The brief pulse of acetylcholine (ACh) released from terminals of the vagus nerve after each impulse can produce two distinct post-synaptic responses in individual principal cells of the ganglion: (i) within a milli-second of release, ACh generates a rapid and strong excitatory post-synaptic potential (e.p.s.p.) that normally initiates a post-synaptic impulse; (ii) this excitation is usually followed by a slow hyperpolarizing inhibitory post-synaptic potential (i.p.s.p.) that lasts for several seconds. The magnitude and time course of the i.p.s.p. depends on the frequency and number of vagal stimuli. When the hydrolysis of ACh is inhibited by prostigmine, a train of nerve stimuli may be followed by an i.p.s.p. lasting half a minute or longer.3. The rapid e.p.s.p. and slow i.p.s.p. result from the direct action of ACh on two different types of chemoreceptors in the post-synaptic membrane of the principal cell. The e.p.s.p. can be preferentially blocked by the nicotinic antagonist dihydro-beta-erythroidine (5 x 10(-7)M), while the i.p.s.p. is selectively blocked by the muscarinic antagonist atropine (5 x 10(-9)M).4. Potentials resembling nerve-evoked e.p.s.p.s and i.p.s.p.s can be produced by iontophoretic release of ACh from micropipettes onto the post-synaptic membrane. Application of the muscarinic agonist bethanechol generates exclusively inhibitory responses.5. The reversal potential for the i.p.s.p. is about -105 mV, which is approximately the equilibrium potential for potassium (E(K)). When the external K(+) concentration is altered, the reversal potential for inhibition is shifted to the new value of E(K) as expected from the Nernst equation. Changes in the external Na(+) and Cl(-) concentrations have no appreciable effect on the reversal potential. Thus, the i.p.s.p. is the result of a conductance increase for K(+).6. The conductance change producing the i.p.s.p. is voltage sensitive. When the membrane potential is shifted from -40 to -60 mV, the i.p.s.p becomes larger and longer. Beyond -60 mV the inhibitory response decreases in proportion to the driving force on K(+) without any further change in time course.7. The inhibitory response produced by an iontophoretically applied pulse of bethanechol has a delayed onset of about 150 msec at 24 degrees C. The early portion of this response, including the delay, is proportional to t(3), where t is time. The proportionality factor (the apparent rate constant) decreases elevenfold when the temperature is lowered by 10 degrees C. This suggests that a multi-step process is involved in the activation of the conductance increase that leads to the inhibitory response. Inhibitory responses with similar kinetics were produced in heart muscles of the mudpuppy upon application of ACh.

Acetylcholine↗

PubMed1970

Receptor potentials from hair cells of the lateral line.

Intracellular recordings from hair cells in the tail lateral line of mudpuppy Necturus maculosus show receptor potentials less than 800 microvolts, peak to peak, from stimuli that are considered large compared to natural stimuli. The hair cells are in neuromasts that are sensitive at the time of recording and are identified by both in vivo and in vitro examination of intracellular staining.

Animals↗

PubMed1972

Cilia: activation coupled to mechanical stimulation by calcium influx.

Ciliated epithelial cells in the oviduct of Necturus maculosus were stimulated mechanically by brief dimpling with a microstylus. This treatment produlced a transient depolarization of the membrane, and a transient increase in the frequency of ciliary beating. The increase in frequency of ciliary beating was related to the concentration of extracellular calcium ion, decreasing with reductiotn in calcium. Addition of lanthanum was followed by a decrease in spontaneous ciliary aictivity and a hyperpolarization of the membrane. In the presence of lanthanum, the transietnt depolarization in response to mechanical stimulation had a shorter timte course, and the concomitant increase in ciliary frequency was greatly reduced. It is concluded that calciuml ions enter the cell as a result of mechanical stimulationi of the membrane, and that calcium influx leads to an increase in the frequency of ciliary activity.

Animals↗