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Bycatch is one of the leading threats facing aquatic organisms, worldwide. Pesticide bycatch can occur when nontarget species are incidentally killed during chemical application targeting noxious species. The common mudpuppy (Necturus maculosus) is a river‐dwelling salamander that is susceptible to bycatch from 3‐trifluoromethyl‐4‐nitrophenol (TFM) applications. However, the long‐term consequences of TFM bycatch on mudpuppy populations are not well understood. We used population viability analysis to reveal potential impacts of TFM application on a mudpuppy population in Ohio's Grand River under plausible bycatch scenarios. We found that the population was more sensitive to increased juvenile mortality and rapid declines occurred when both adults and juveniles were impacted at high bycatch rates (≥5% of the population) or more frequent TFM intervals (≥5 years). Our results suggest that continued use of TFM could pose a risk to mudpuppy population persistence if bycatch occurs at moderate to high levels and affects both juveniles and adults. Future TFM use and management decisions should be directed with these potential nontarget impacts in mind. [ABSTRACT FROM AUTHOR]

In March 2017, a wild‐caught female common mudpuppy Necturus maculosus from Iowa, USA, with an enlarged posterior abdomen was submitted for diagnostic assessment. The cause of the abdominal distension was a large fluid‐filled abdominal mass, diagnosed as a nephroblastoma. Parasites and numerous bacteria were isolated and identified from the mudpuppy but were determined to be incidental. Samples of the neoplasm inoculated onto an American toad Anaxyrus americanus cell line (BufoTad) yielded cytopathic effect during several passages. However, standard molecular testing of the cell culture supernatant failed to identify any viruses. Next‐generation sequencing identified the replicating agent as a bacterium of the genus Acholeplasma. Immunohistochemistry confirmed the presence of Acholeplasma within the nephroblastoma, including within tumor cells. This is the first report of nephroblastoma and the second report of neoplasia in this species. The results also suggest that certain bacteria of the genus Acholeplasma might be oncogenic. [ABSTRACT FROM AUTHOR]

During a routine health inspection of apparently healthy wild‐caught common mudpuppies Necturus maculosus, the bacteria Yersinia ruckeri was isolated and the identity confirmed using biochemical and molecular methods. This represents the first isolation of Y. ruckeri from an amphibian. This finding increases the known host range capable of harboring this important fish pathogen and could have serious management implications for aquaculture. Furthermore, addressing wild amphibians in fish hatchery biosecurity plans is discussed. [ABSTRACT FROM AUTHOR]

A major goal of genomic and reproductive biology is to understand the evolution of sex determination and sex chromosomes. Species of the 2 genera of the Salamander family Proteidae - Necturus of eastern North America, and Proteus of Southern Europe - have similar-looking karyotypes with the same chromosome number (2n = 38), which differentiates them from all other salamanders. However, Necturus possesses strongly heteromorphic X and Y sex chromosomes that Proteus lacks. Since the heteromorphic sex chromosomes of Necturus were detectable only with C-banding, we hypothesized that we could use C-banding to find sex chromosomes in Proteus . We examined mitotic material from colchicinetreated intestinal epithelium, and meiotic material from testes in specimens of Proteus, representing 3 genetically distinct populations in Slovenia. We compared these results with those from Necturus . We performed FISH to visualize telomeric sequences in meiotic bivalents. Our results provide evidence that Proteus represents an example of sex chromosome turnover in which a Necturus -like Y-chromosome has become permanently translocated to another chromosome converting heteromorphic sex chromosomes to homomorphic sex chromosomes. These results may be key to understanding some unusual aspects of demographics and reproductive biology of Proteus, and are discussed in the context of models of the evolution of sex chromosomes in amphibians. [ABSTRACT FROM AUTHOR]

The Mudpuppy (Necturus maculosus) is an entirely aquatic salamander whose geographic range is thus defined by immigration routes in watersheds that permit feasible travel. Significant barriers, such as large waterfalls, effectively bar this species from further colonization upstream. We compared the contemporary distribution of Mudpuppies in southern Ontario with varying post-glacial ancient lake maxima and riverine outlet-flow conditions. Topography does not appear to be a range-limiting factor, but the type of river grade (waterfalls versus riffles) does. The distribution of modem records of this species in Ontario aligns closely with maxima from the Nipissing phase occurring 4000-5000 years ago, leading us to suggest that this is when Mudpuppies invaded and proliferated in the Great Lakes Basin. [ABSTRACT FROM AUTHOR]

Although gap junctions are widely expressed in the developing central nervous system, the role of electrical coupling of neurons and glial cells via gap junctions in the spinal cord in adults is largely unknown. We investigated whether gap junctions are expressed in the mature spinal cord of the mudpuppy and tested the effects of applying gap junction blocker on the walking-like activity induced by NMDA or glutamate in an in vitro mudpuppy preparation. We found that glial and neural cells in the mudpuppy spinal cord expressed different types of connexins that include connexin 32 (Cx32), connexin 36 (Cx36), connexin 37 (Cx37), and connexin 43 (Cx43). Application of a battery of gap junction blockers from three different structural classes (carbenexolone, flufenamic acid, and long chain alcohols) substantially and consistently altered the locomotor-like activity in a dose-dependent manner. In contrast, these blockers did not significantly change the amplitude of the dorsal root reflex, indicating that gap junction blockers did not inhibit neuronal excitability nonselectively in the spinal cord. Taken together, these results suggest that gap junctions play a significant modulatory role in the spinal neural networks responsible for the generation of walking-like activity in the adult mudpuppy. [ABSTRACT FROM AUTHOR]

This study used an in situ heart preparation to analyze the power output and stroke work of spontaneously beating hearts of four anurans ( Rhinella marina, Lithobates catesbeianus, Xenopus laevis, Pyxicephalus edulis) and three urodeles ( Necturus maculosus, Ambystoma tigrinum, Amphiuma tridactylum) that span a representative range of relative ventricle mass (RVM) found in amphibians. Previous research has documented that RVM correlates with dehydration tolerance and maximal aerobic capacity in amphibians. The power output (mW g ventricle mass) and stroke work (mJ g ventricle muscle mass) were independent of RVM and were indistinguishable from previously published results for fish and reptiles. RVM was significantly correlated with maximum power output ( P, mW kg body mass), stroke volume, cardiac output, afterload pressure ( P) at P, and preload pressure ( P) at P. P at P and P at P also correlated very closely with each other. The increases in both P and P at maximal power outputs in large hearts suggest that concomitant increases in blood volume and/or increased modulation of vascular compliance either anatomically or via sympathetic tone on the venous vasculature would be necessary to achieve P in vivo. Hypotheses for variation in RVM and its concomitant increased P in amphibians are developed. [ABSTRACT FROM AUTHOR]

We performed an exploratory analysis of the morphology of the cranium in the white olm (Proteus anguinus anguinus) and the black olm (P. a.parkelj) with micro-CT scanning and geo- metric morphometrics. The mudpuppy (Necturus maculosus) was used as an outgroup. The black olm falls outside the white olm morphospace by a markedly wider skull, shorter vomers which are positioned further apart and by laterally positioned squamosals and quadrates relative to the palate (the shape of the buccal cavity). On account of its robust skull with more developed premaxillae a shorter otico-occipital region, the black olm is positioned closer to Necturus than are the studied specimens of the white olm. The elongated skull of the white olm, with an anteriorly positioned jaw articulation point, could be regarded as an adaptation for improved feeding success, possibly compensating for lack of vision. As yet, the alternative explanations on the evolution of troglomorphism in Proteus are an extensive convergence in white olms versus the reverse evolution towards less troglomorphic character states in the black olm. To further understand the evolutionary trajectories within Proteus we highlight the following hypotheses for future testing: i) morphological differentiation is smaller within than between genetically differentiated white olm lineages, and ii) ontogenetic shape changes are congruent with the shape changes between lineages. We anticipate that the morphological detail and analytical power that come with the techniques we here employed will assist us in this task. [ABSTRACT FROM AUTHOR]

Amphibian liver pigment cells, also known as Kupffer cells, contain various amounts of melanin, haemosiderin and lipofuscin. We used different histochemical and ultrastructural methods to analyse and compare the level of hepatic pigmentation and the structure of hepatic pigment cells in the livers of three representatives of the family Proteidae; two subspecies of the hypogean Proteus anguinus (depigmented Proteus a. anguinus and pigmented Proteus a. parkelj) and the epygean Necturus maculosus. Our analysis revealed differences at histological and ultrastructural level. While the percentage of the pigmented area and ultrastructural characteristics are similar in both subspecies of P. anguinus, great differences occur in the amount and structure of the pigment cells between P. anguinus and N. maculosus. Pigment cells are more numerous and compose larger pigmented clusters in P. anguinus. They are structurally more heterogeneous and contain a larger amount of haemosiderin when compared to N. maculosus. Our results confirm a high degree of variation in hepatic pigmentation among different amphibian species. Because many factors influence the level of hepatic pigmentation in poikilotherms, differences among species from different habitats and also among individuals of the same species are expected but are not easily explained. We propose two possible explanations for the large amount of iron present in Proteus anguinus: (1) accumulation of pigments due to the very low metabolic rate and extended lifetime; (2) large iron storage capacity as an adaptation to a low and discontinuous food supply in caves. [ABSTRACT FROM AUTHOR]

Heterochrony, a difference in developmental timing, is a central concept in modern evolutionary biology. An example is pedomorphosis, retention of juvenile characteristics in sexually mature adults, a phenomenon largely represented in salamanders. The mudpuppy ( Necturus maculosus) is an obligate pedomorphic amphibian, never undergoing metamorphosis. Thyroid hormone induces tissue transformation in metamorphosing species and this action is mediated by nuclear thyroid hormone (TH) receptors (TRs). The absence of metamorphosis in Necturus has been attributed to a resistance to TH action as treatment with exogenous TH fails to induce transformation. The failure to metamorphose could be due to the lack of TR expression in target tissues, or to a loss of TR function. Toward understanding the molecular basis for the failure of Necturus tissues to respond to TH, and the ultimate cause for the expression of the obligate pedomorphic life history, we characterized the structure, function, and expression of TR genes in Necturus. Strikingly, we found that Necturus TRα and TRβ genes encode fully functional TR proteins. These TRs bind both DNA and TH and can transactivate target genes in response to TH. Both TRα and TRβ are expressed in various tissues. TH treatment in vivo induced expression in the gill of some but not all genes known to be activated by TH in anuran larvae, caused whole organism metabolic effects, but induced no external morphological changes in adults or larvae. Thus, Necturus possesses fully functional TRs and its tissues are not generally resistant to the actions of TH. Rather, the absence of metamorphosis may be due to the loss of TH-dependent control of key genes required for tissue transformation. [ABSTRACT FROM AUTHOR]

Many odor responses are mediated by the adenosine 3′,5′-cyclic monophosphate (cAMP) pathway in which the cAMP-gated current is amplified by Ca2+-dependent Cl− current. In olfactory neurons, prolonged exposure to odors decreases the odor response and is an adaptive effect. Several studies suggest that odor adaptation is linked to elevated intracellular Ca2+. In the present study, using the perforated configuration of the patch clamp technique, we found that repetitive odor stimulation elicits a potentiation of the subsequent responses in olfactory neurons. This potentiation is mimicked by stimulating the cAMP pathway and does not appear to be related to phosphorylation of ion channels since protein kinase inhibitors could not block it. Our data suggest that local increases in [Ca2+]i via activation of the cAMP pathway mediate the pulse-elicited potentiation. In the first odor application, entry of Ca2+ through cyclic nucleotide–gated channels appears to be buffered. Repetitive stimulation allows local increases in [Ca2+]i, recruiting more Ca2+-dependent Cl− channels with each subsequent odor pulse. [ABSTRACT FROM AUTHOR]

The isolated brachial spinal cord of the mudpuppy is useful for studies of neural networks underlying forelimb locomotion, but information about its anatomy is scarce. We addressed this issue by combining retrograde labeling with fluorescent tracers and confocal microscopy. Remarkably, the central region of gray matter was aneural and contained only a tenuous meshwork of glial fibers and large extracellular spaces. Somata of motoneurons (MNs) and interneurons (INs), labeled retrogradely from ventral roots or axons in the ventro-lateral funiculus, respectively, were confined within a gray neuropil layer abutting the white matter borders, while their dendrites projected widely throughout the white matter. A considerable fraction of labeled INs was found contralaterally with axons crossing beneath a thick layer of ependyma surrounding the central canal. Dorsal roots (DRs) produced dense presynaptic arbors within a restricted dorsal region containing afferent terminations, within which dorsally directed MN and IN dendrites mingled with dense collections of synaptic boutons. Our data suggest that a major fraction of synaptic interactions takes place within the white matter. This study provides a detailed foundation for electrophysiological experiments aimed at elucidating the neural circuits involved in locomotor pattern generation. [ABSTRACT FROM AUTHOR]

Recently collected specimens document the presence of the Mudpuppy at the mouths of tributaries draining into the tidal Hudson Rivet in Columbia and Greene Counties. NY. In this study, we summarize historic records of Mudpuppy from the Hudson River. Examination of these records, Mudpappy distribution elsewhere, and the distribution of riverine New York reptiles leads us to conclude that Mudpuppy should be considered native to the Hudson Valley. [ABSTRACT FROM AUTHOR]

The presence of metallothionein (MT) and the subcellular distribution of copper, zinc and cadmium were investigated in livers of two neotenic salamanders, Proteus anguinus and Necturus maculosus. In P. anguinus, caught in the wild, hepatic MTs were present as a single isoform of (Zn, Cu, Cd)-thioneins, whose molecular weight was estimated to be approximately 12 000 by size exclusion chromatography. The percentage of zinc and cadmium was higher in the cytosol and of copper in the pellet. Cytosolic cadmium was almost exclusively associated with MTs (80%), while zinc and copper were also present in the regions of higher-molecular weight proteins. In laboratory bred N. maculosus, MTs were isolated from the liver cytosol and extract of the pellet as (Cu, Zn)- and (Zn, Cu)-thioneins, respectively. According to the low amount of copper extracting from liver pellets of N. maculosus, the presence of water insoluble aggregated forms of Cu-thioneins should be checked in further investigations. [Copyright &y& Elsevier]

Odor transduction mediated by the adenylyl cyclase/cAMP pathway has been well studied, but it is still uncertain whether this pathway mediates the transduction of all odors in vertebrates. We isolated olfactory sensory neurons from the salamander Necturus maculosus and used calcium imaging with the indicator dye fura-2 to examine olfactory responses elicited by amino acids. The properties of approximately two-thirds of the odor responses suggested they were mediated by the adenylyl cyclase/cAMP pathway, but one-third of the responses were not mimicked by cAMP analogs nor blocked by inhibition of adenylyl cyclase, suggesting that these odor responses were mediated differently. Responses that were unaffected by inhibition of adenylyl cyclase were blocked by neomycin, an inhibitor of phospholipase C, implying that they were transduced by activation of phospholipase C. Some cells which responded to more than one amino acid appeared to employ both pathways, but each was used to transduce different odors. In addition, many responses that were mediated by the adenylyl cyclase/cAMP pathway were enhanced following inhibition of phospholipase C, suggesting that the phospholipase C pathway has a role not only in odor transduction, but also in the modulation of olfactory responses. [ABSTRACT FROM AUTHOR]

It is well established, for all Na-absorbing epithelia, that an increase in the rate of transcellular Na+ absorption is accompanied by an increase in the conductance of the basolateral membrane to K+. For the case of small intestinal epithelial cells from the salamander Necturus maculosus, where the rate of transcellular Na+ absorption can be increased manyfold by the addition of sugars or amino acids to the luminal bathing solution, it appears that this parallelism between Na-K pump rate and basolateral membrane K+ conductance is closely related to volume regulation by the enterocyte. Recent studies have disclosed the presence of stretch-activated K+ channels, in a highly enriched basolateral membrane fraction isolated from these epithelial cells, whose activity is increased by an increase in vesicle volume and inhibited by a decrease in vesicle volume or ATP. The activity of this channel also appears to be regulated by the degree of organization of the cortical actin cytoskeleton; activity is increased by depolymerization of the actin cytoskeleton and decreased by repolymerization of that structure. We postulate that the inhibitory effect of ATP is related to its role in promoting the polymerization of G-actin to form F-actin. We propose that enterocyte swelling that results from the intracellular accumulation of sugars or amino acids in osmotically active forms brings about disorganization of the cortical actin cytoskeleton and activates these channels and is, at least in part, responsible for the "pump-leak parallelism" in this amphibian. [ABSTRACT FROM AUTHOR]

Presents a study which compared how hydrochloric acids and other organic acids affect the potential of hydrogen ions in the gastric mucosa epithelial cells of Necturus maculosus. Methodology; Results; Discussion.

Examines the effect of increased potassium[sup +] activity of membrane vesicles of Necturus maculosus intestinal cells. Decline in local adenosine triphosphate (ATP) activity; Induction of stretch-activated potassium[sup +] (SAK) channels by cell swelling; Decrease of the inhibitory effect of ATP on the SAK channels.

Carbonic anhydrase (CA) activity in blood and other tissues and red blood cell (rbc) anion exchange were measured in the mud puppy, Necturus maculosus, in order to gain insight into the strategy for CO[sub2] transport used by these neotenic salamanders and to further explore evolutionary relationships between rbc CA activity and anion exchange in nonmammalian vertebrates. CA activity was detectable in all of the tissues examined, but CA activity in blood was much lower than that in most vertebrates. There was no indication, however, that additional CA had been incorporated into the membrane fraction of other tissues to compensate for this low blood CA activity. In further contrast to most other animals, low levels of CA activity were also detectable in mud puppy plasma. Preliminary characterization of the rbc CA indicated that the Type II, fast-turnover enzyme was indeed present, but that there are a very low number of active sites in mud puppy rbc's. Further experiments showed that the rbc's were highly permeable to anions and that the relative rate of anion flux could be inhibited by 4,4-diisothiocyanostilbene-2,2-disulphonic acid. Thus, the process of CO[sub2] transport in the blood of mud puppies probably involves components of the Jacobs-Stewart cycle, as in most other vertebrates. [ABSTRACT FROM AUTHOR]

Examines whether extracellular adenosine triphosphate (ATP) stimulates regulatory volume decrease in Necturus maculosus red blood cells. Reversal of the inhibitory effect of apyrase with the nonhydrolyzable analog adenosine 5'-O-(3-thiotriphosphate); Indication that extracellular ATP stimulated cell volume decrease via a calcium-dependent step that led to potassium efflux.

Reports on the functional characteristics of electrically coupled and noncoupled taste bud cells from the lingual epithelia prepared from Necturus maculosus using the patch clamp technique. Two distinct taste cell subpopulations based on their electrophysiological properties; Voltage-gated Na+, K+ and Ca+2 currents within receptor cells.

Focuses on a study which examined the possibility that KATP channel in the basolateral membrane of Necturus maculosa enterocytes can be affected by ATP produced locally from glycolytic substrates. Materials and methods; Adenylate kinase activity; Activities of pyruvate kinase, lactic dehydrogenase and coupled system.

Describes a method to isolate epithelial cells from gallbladders of Necturus maculosus with preserved structural and functional polarity. Cell labeling with fluorescent probes; Antibodies and fluorescent probes; Cell isolation; Confocal fluorescence microscopy; Measurements of cell membrane voltage.

Evaluates the changes in electrophysiological and secretory properties of surface epithelium of the gastric antrum of the Necturus maculosus in animals. Use of classical ion flux measurements and intracellular microelectrode techniques; Materials and methods used in experiments; Findings; References.

Describes a preparation of necturus maculosus gallbladded epithelium yielding isolated cells that retain structural and functional polarity. Morphology of isolated epithelial cells; Immunodetection of a CFTR homologue; Electrophysiological techniques.

Previously 2D cable analysis has been performed with two microelectrodes, one for passing intraepithelial current and the other for measuring the voltage response in multiple cells along the distance scale. This requires that the epithelium must be in a stable state for a considerably long period because of the multiple impalements. To follow changes of intraepithelial resistances in chambered Necturus antral mucosa with good temporal resolution, four/five electrodes were used to impale cells in the same preparation and the intraepithelial current (6.5–20 nA) was conducted sequentially to three/four of the electrodes, one at a time, to obtain six/ten independent voltage response measurements along the distance scale. The solution to the 2D cable equations was fitted to results and apical, basolateral and shunt resistances were calculated. It was found that an incorrect distance configuration can ruin the precision of the measurement. The distance configuration can, however, be optimized. The resistance values calculated with the 2D cable analysis were very close to those obtained by the amiloride exposure technique in the same tissues. The improvement gained with this work is better temporal resolution (even <10 s) when measuring epithelial resistances. [ABSTRACT FROM AUTHOR]

The standard metabolic rate (SMR) and critical O tension (P) of water-breathing mudpuppies and hellbenders were determined at 20° C using open-system respirometry. Both species are metabolic O regulators, although the P of hellbenders (90 mmHg) is much higher than that of mudpuppies (40 mmHg). The SMR of the two species in water saturated with air was similar (19.5 and 20.0 μl O/g·h for Cryptobranchus and Necturus, respectively) and not different from that of salamanders in general. Both species were able to survive for at least 5-11 days in severely hypoxic water (9-10 mmHg) by breathing air, indicating that the lungs are functional accessory respiratory structures. We conclude that hellbenders are restricted to relatively cool and flowing waters because of their limited gas exchange capabilities, particularly with regard to their limited aerobic scope for activity and slow recovery from exercise. Necturus maculosus is much more tolerant of hypoxia, but it is not known if they can inhabit areas were hypoxia is combined with hypercarbia. [ABSTRACT FROM AUTHOR]

This study examined whether protein kinase C (PKC) stimulates K+ efflux during regulatory volume decrease (RVD) in Necturus maculosus (mudpuppy) red blood cells (RBCs). The limit of osmotic fragility increased with the general protein kinase inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7, 10 micrometer), but not with the cyclic nucleotide-dependent kinase antagonists N-(2'-guanidinoethyl)-5-isoquinolinesulfonamide (HA-1004, 10 micrometer) and N-2-(methylamino)ethyl-5-isoquinoline-sulfonamide (H-8, 5 micrometer). Consistent with these results, osmotic fragility also increased with the PKC antagonists bisindolylmaleimide I (GF-109203X or bis I, 100 nm), bisindolylmaleimide II (bis II, 100 nm), and chelerythrine (10 micrometer). The effect of these three antagonists and H-7 was reversed with gramicidin (5 micrometer in a choline Ringer), indicating PKC was linked to K+ efflux (gramicidin is a cationophore that was used to ensure a high K+ permeability). We also measured cell volume recovery from hypotonic shock (0.5x Ringer) with a Coulter counter and estimated cell volume from the hematocrit. The percent RVD compared to control decreased with H-7 (10 micrometer), sphingosine (100 nm), chelerythrine (10 micrometer), bis I (100 nm), and bis II (100 nm), but not with HA-1004 (10 micrometer) nor H-8 (5 micrometer). Inhibition of RVD by H-7, chelerythrine, bis I, and bis II was reversed with gramicidin (5 micrometer). Furthermore, using the patch clamp technique, we found H-7 (10 micrometer) reduced a whole cell conductance that was activated during cell swelling. In addition, a conductance responsible for K+ efflux during cell swelling was inhibited by bis I (100 nm) and bis II (100 nm). These results indicate that a conductive pathway mediating K+ loss during RVD is regulated, at least in part, by protein kinase C. [ABSTRACT FROM AUTHOR]

We examined whether metabolites of arachidonic acid (AA) regulate K+ efflux during regulatory volume decrease (RVD) by mudpuppy red blood cells (RBCs). Volume regulation was inhibited by the phospholipase A2 antagonists mepacrine (10 microM) and ONO-RS-082 (10 microM); the inhibitory effect of ONO-RS-082 was reversed by gramicidin (5 microM). Eicosatetraynoic acid (ETYA, 100 microM), a general antagonist of AA metabolism, also blocked RVD. In addition, volume regulation was inhibited by the lipoxygenase pathway antagonist nordihydroguaiaretic acid (NDGA, 10 microM), the 5 lipoxygenase antagonists AA-861 (5 microM) and curcumin (20 microM), and by the 5-lipoxygenase activating protein inhibitor L-655,298 (5 microM). Inhibition by all four of these agents was reversed with gramicidin. In contrast, the 12- and 15-lipoxygenase pathway inhibitor ethyl-3,4-dihydroxy-benzylidene-cyanoacetate (EDBCA, 1 microM) and the cytochrome P-450 monooxygenase pathway blocker ketoconazole (20 microM) had no effect. On the other hand, the cyclooxygenase pathway inhibitor aspirin (100 microM) slightly enhanced RVD. Consistent with these findings, a K(+)-selective whole cell conductance responsible for K+ efflux during cell swelling was inhibited by ONO-RS-082 (10 microM), NDGA (10 microM), AA-861 (5 microM), curcumin (20 microM), and L-655,298 (5 microM). In contrast, EDBCA (1 microM), ketoconazole (20 microM), and indomethacin (10 microM) did not block this whole cell conductance. These results indicate that a channel mediating K+ loss during RVD is regulated by a 5-lipoxygenase metabolite of arachidonic acid. [ABSTRACT FROM AUTHOR]

Early diplotene oocytes from Necturus maculosus ranging from ∼ 0.2 to 0.5 mm in diameter were examined by electron microscopy. In the smallest oocytes of this range, the cytoplasm is largely devoid of membranes, but contains primarily ribosomes and mitochondria. In slightly larger oocytes, smooth-surfaced cytomembranes first appear in the perinuclear cytoplasm. At this time, the outer layer of the germinal vesicle nuclear envelope (GVNE) shows frequent connections with long membranous lamellae that extend for considerable, but variable distances into the juxtanuclear ooplasm. The number of smooth membranous lamellae increases tremendously as the oocytes increase in diameter. In such oocytes as well, frequent continuities are observed between the outer membrane of the GVNE and many of the cytoplasmic membranes. Eventually, as the ooplasm becomes populated with extensive numbers of membranous lamellae, instances of continuity between the membranous lamellae and nuclear envelope now become sparse and eventually non-existent. The frequent connections observed between membranous lamellae and the outer membrane of the GVNE during a circumscribed interval of diplotene strongly implicate the GVNE in the generation of extensive amounts of cytoplasmic membrane. The ooplasm of larger oocytes in the size range indicated contain numerous Golgi complexes and large quantities of annulate lamellae most of which are positioned in the peripheral or subcortical ooplasm, as well as extensive quantities of smooth membranes of the endoplasmic reticulum and lipid droplets. [ABSTRACT FROM AUTHOR]

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: (1) the absence of a well defined moulting cycle together with an apparently diminished synthetic and mitotic activity in the stratum germinativum; (2) an outermost cell layer ( stratum mucosum) that is unkeratinized and appears to synthesize a mucous layer; and (3) 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. [ABSTRACT FROM AUTHOR]

Neurons accumulating (H)-glycine and (H) GABA were demonstrated with the use of autoradiography. Both were accumulated by different types of amacrine cells, similar those of goldfish. (H)-GABA was also accumulated by horizontal cells, again similar to the goldfish. These results and physiological studies from other laboratories suggest that GABA and glycine are neurotransmitter candidates in amacrine cells of the mudpuppy. Immunoreactive neuropeptide Y (NPY), glucagon, vasoactive intestinal peptide (VIP), somatostatin, substance P, and neurotensin were found in different types of stratified amacrine cells. Weakly immunoreactive enkephalin and bombesin processes were also seen in the inner plexiform layer. Gastrin-immunoreactive neurons were not detectable. Endogenous 5-hydroxytryptamine was visualized immunohistochemically in a population of diffuse amacrine cells and some cells in the ganglion cell layer. This suggests that 5-hydroxytryptamine may be a neurotransmitter in the retina of the mudpuppy. [ABSTRACT FROM AUTHOR]