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PubMed2005

Status of RNAs, localized in Xenopus laevis oocytes, in the frogs Rana pipiens and Eleutherodactylus coqui.

Early development in the frog model, Xenopus laevis, is governed by RNAs, localized to the vegetal cortex of the oocyte. These RNAs include Xdazl RNA, which is involved in primordial germ cell formation, and VegT RNA, which specifies the mesoderm and endoderm. In order to determine whether orthologues of these RNAs are localized and have similar functions in other frogs, we cloned RpDazl and RpVegT from Rana pipiens, a frog that is phylogenetically distant from X. laevis. RNAs from both genes are localized to the vegetal cortex of the R. pipiens oocyte, indicating that the vegetal localization is likely the basal state. The animal location of EcVegT RNA in Eleutherodactylus coqui that we found previously (Beckham et al., 2003) is then a derived state, probably due to the great increase in egg size required for direct development of this species. To answer the question of function, we injected RpVegT or EcVegT RNAs into X. laevis embryos, and assayed animal caps for gene expression. Both of these RNAs induced the expression of endodermal, mesodermal, and organizer genes, showing that the function of RpVegT and EcVegT as meso-endodermal determinants is conserved in frogs. The RNA localizations and the function of VegT orthologues in germ layer specification may be synapomorphies for anuran amphibians.

Amino Acid Sequence↗
PubMed2005

Protamines in the internally fertilizing neobatrachian frog Eleutherodactylus coqui.

The internally fertilizing primitive frog Ascaphus truei (family Ascaphidae) from the Pacific Northwest is the only frog with an intromittent organ. The more advanced neobatrachian frog Eleutherodactylus coqui (family Leptodactylidae) from Puerto Rico has secondarily acquired internal fertilization but mates by cloacal apposition. Nonetheless, both frogs have introsperm with an elongated head containing highly condensed chromatin. Characterization of sperm nuclear basic proteins (SNBPs) in E. coqui by acid-urea polyacrylamide gel electrophoresis indicates that, as in A. truei, testes from a single animal contain several protamines. Amino acid analysis indicates a composition for the most rapidly moving protamine of each species as follows: in E. coqui, ARG (35.6 mol %) + LYS (3.8 mol %) + HIS (7.6 mol %) = 47 mol % total basic residues and in A. truei, ARG (42.1 mol %) + LYS (11.1 mol %) = 53.2 mol % total basic residues. Transmission electron microscopy shows that E. coqui introsperm, like those in A. truei, are elongate with highly condensed chromatin. However, E. coqui introsperm lacks an axial perforatorium that extends into an endonuclear canal. These morphological features are plesiomorphic (primitive) and shared by A. truei with urodeles and basal amniotes (Jamieson et al. (1993) Herpetologica 49:52-65). In E. coqui introsperm, the nucleoprotein complex has a cross-sectional axis of 420 + 20 angstroms and shows a knobby chromatin structural organization in TEM. The presence of arginine-enriched protamines in both a basal anuran like the ascaphid A. truei and a more advanced neobatrachian like the leptodactylid E. coqui supports the hypothesis that internal fertilization acts as a constraint on the range of SNBP diversity in animals.

Amino Acids↗
PubMed1995

Sequence analysis of frog alpha-crystallin cDNA and its deduced primary structure: comparison of alpha A subunit chains among different vertebrate species.

alpha-Crystallin which is present in the lenses of all vertebrate species is a major lens protein with blocked amino terminus. To facilitate the cloning and sequence determination of amphibian alpha-crystallin, total cDNA mixture was constructed from the poly(A)+mRNA of eye lenses from the bullfrog. cDNAs encoding alpha A and alpha B chains of alpha-crystallin were then amplified by polymerase chain reaction (PCR) using primers based on the N- and C-terminal amino-acid sequences of conserved mammalian alpha-crystallin subunit chains. PCR-amplified product corresponding to alpha A crystallin subunit was obtained, which was then subcloned into pUC18 vector and then transformed into E. coli strain JM109. Plasmids purified from the positive clones were prepared for nucleotide sequencing by dideoxynucleotide chain-termination method. Sequencing several clones containing DNA inserts coding for alpha A-crystallin constructed a complete full-length reading frame of 522 base pairs covering a deduced protein sequence of 173 amino acids including the universal translation-initiating methionine. The frog alpha A shows 95, 82 and 81% sequence identity to alpha A crystallin chains of another species of Rana frog, bovine and human, respectively, revealing the close relationship among alpha A crystallins from remotely related species. The present report on the determination of primary structure for frog alpha A-crystalline chain through cDNA cloning and sequencing has provided the missing N-terminal segment of alpha A sequence reported for another amphibian species and would represent the first complete frog alpha A-crystallin sequence solved by PCR technique.

Amino Acid Sequence↗
PubMed1995

Isolation of ice-nucleating active bacteria from the freeze-tolerant frog, Rana sylvatica.

Ice-nucleating active (INA) bacteria were isolated from the gut of field-collected freeze-tolerant wood frogs (Rana sylvatica) collected in winter. Thirteen strains of Pseudomonas fluorescens, four strains of Pseudomonas putida, and two strains of Enterobacter agglomerans had ice-nucleating activity. Each of the INA pseudomonad strains was psychrophilic. P. putida strains were differentiated from P. fluorescens strains by gelatinase, lecithinase, and lipase production. The maximum nucleation temperatures (Tmax) of aqueous suspensions (10(9) bacteria/ml) of the four INA P. putida strains ranged from -1.6 to -3.0 degrees C, which places this INA species among the most potent known biological nucleators. Ingestion of INA P. putida isolated from R. sylvatica by another freeze-tolerant frog. Pseudacris crucifer, decreased the capacity of this frog to supercool and remain unfrozen at -2 degrees C. This is the first report of INA bacteria isolated from a vertebrate, and suggests that, as part of the gut flora in some posthibernation freeze-tolerant wood frogs, these bacteria may play a role in enhancing winter survival by promoting ice nucleation at high subzero temperatures (ca. -2 degrees C).

Animals↗
PubMed1995

Receptor-mediated endocytosis of insulin in lower vertebrates: internalization and intracellular processing of 125I-insulin in isolated hepatocytes of lamprey and frog.

The binding of 125I-insulin to cellular insulin receptors and the internalization of insulin-receptor complexes have been studied in isolated hepatocytes of frog and lamprey. Two classes of binding sites (Kd 10(-9) and 10(-8) M) were found in cells of both species. The molecular weight of the insulin receptor alpha-subunit was 130 kDa in both species. Internalization of bound 125I-insulin in both species was found in the temperature range 0 to 20 degrees. Cells "loaded" with 125I-insulin were used to estimate the fate of the internalized ligand. Release of internalized ligand from frog cells increased at temperatures ranging from 0 to 20 degrees. At 0 degrees the degraded 125I-insulin was 5%, at 5 degrees 7%, and at 20 degrees 17% of total radioactivity accumulated in the medium. In lamprey hepatocytes there was neither radioactivity accumulation in the incubation medium nor release from cells at all temperatures studied. The intracellular degradation of internalized 125I-insulin in frog hepatocytes was much lower than that in lamprey cells. In frog hepatocytes the specific binding of 125I-insulin was increased twofold in the presence of the lysosomal inhibitor chloroquine. In contrast no increase was found in lamprey hepatocytes. In conclusion, the processing pathways of internalized insulin in the cells of ectothermal and endothermal vertebrates are generally similar but in ectothermal animals all events take place at lower temperatures and at lower rates. The peculiarities of insulin processing in lamprey hepatocytes most likely result from the transformation of hepatocytes during the nonfeeding prespawning period.

Animals↗
PubMed1996

The role of androgens in opportunistic breeding, tropical frogs.

In this paper we present the results of an interspecific study on the androgen levels of several species of Old World tropical frogs. These data are used to test three hypotheses: (1) in unpredictable, aseasonal environments with opportunistic breeding, gonadal hormones have a permissive rather than an activating role in the expression of mating behavior (Crews and Moore, 1986), (2) a behaviorally induced androgen response is present in opportunistic aseasonal species as well as in temperate seasonal breeders (Crews and Moore, 1986; Wingfield et al., 1990), and (3) because maintenance of high plasma testosterone levels has an associated energetic and/or fitness cost (Moore and Marler, 1987; Wingfield et al., 1990), continuously breeding frogs have lower testosterone levels than seasonal, discontinuous breeders. Androgen and corticosterone levels were found to vary significantly among species exhibiting the same mating behavior, providing support for the hypothesis that hormones have a permissive role. Differences in androgen level were correlated with (1) variation in level of male-male aggression, (2) relative density of male frogs, and (3) call rate and power output of male vocalization. These results indicate that a behaviorally induced androgen response is present in opportunistic, aseasonal breeders. Furthermore, tropical, aseasonal, opportunistic breeding frogs appeared to have lower androgen levels, generally, than confamilial temperate, seasonal breeders. This difference is consistent with the hypothesis that maintenance of high plasma testosterone levels is energetically expensive.

Androgens↗
PubMed1998

Responses of frog skin, Rana pipiens, calcium ion transport to parathyroid hormone, calcitonin, and vitamin D3.

The responses of cutaneous Ca2+ transport to parathyroid hormone, calcitonin, and vitamin D3 were evaluated in intact Rana pipiens and isolated skins from this species. Parathyroid hormone (12 pmol g-1) stimulated Ca2+ influx in intact frogs but not in isolated skin (10-100 pmol ml-1) unless the frogs had been pretreated with vitamin D3 injections (100-300 microgram day-1) for 3 days prior to isolating the skins for flux measurements. Pretreatment of intact frogs with vitamin D3 for 3 days (100 microgram day-1) also resulted in greater responses to PTH (6-12 pmol g-1). Vitamin D3 (most likely after conversion to 1,25(OH)2D3) had a direct stimulatory effect on Ca2+ transport in isolated skins after three daily doses of 500 microgram of D3. Calcitonin also stimulated Ca2+ influx in both intact frogs (24 pmol g-1) and isolated skins (25-100 pmol ml-1). We conclude that parathyroid hormone, facilitated by vitamin D, stimulates Ca2+ influx in the skin of R. pipiens. We also conclude that calcitonin is stimulatory for cutaneous Ca2+ influx in this species.

Animals↗
PubMed2000

Pattern of gonadal sex differentiation, development, and onset of steroidogenesis in the frog, Rana curtipes.

Histomorphological changes and steroidogenic potential of the gonads during sexual differentiation and development were studied in Rana curtipes from tadpole stage 25 (Gosner) until maturity. In stage 25 tadpoles of smaller snout-vent length (SVL; 4-5 mm) the gonads were indifferent, containing a few somatic and germ cells, whereas in larger tadpoles (SVL > 7 mm) gonads were differentiated into ovaries with a central lumen. Onset of meiosis was seen in these ovaries. At stage 26, diplotene and first growth phase oocytes were found. With advancement in developmental stage and after metamorphosis the ovaries progressively enlarged due to increase in number and size of oocytes. Vitellogenesis began in the ovary of 4-month-old frogs. Females attained maturity 6 months after metamorphosis. The frogs showed amplexus and one frog spawned. Onset of testicular formation seen at stage 31 was associated with the degeneration of oocytes and infiltration of darkly stained somatic cells in the central region. By stage 35 all oocytes degenerated, leaving behind a large number of somatic and germ cells interspersed with each other. At stage 38, formation of seminiferous tubules enclosing spermatogonia and pre-Sertoli cells was seen. Initiation of meiosis was evident at metamorphic climax. Cysts of elongated spermatids associated with Sertoli cells were seen in 45-day-old frogs. Histochemically, delta(5)-3 beta-hydroxysteroid dehydrogenase activity was localized in the ooplasm, follicular cells, and interstitium of the ovary from stage 28 onward. The enzyme activity in the testis appeared in 45-day-old froglets. In R. curtipes gonadal differentiation is a semidifferentiated type since gonads initially differentiate into ovaries, and later, in the prospective males, the ovaries degenerate and transform into testes. The males attain maturity much earlier than the females. In R. curtipes gonadal sex differentiation precedes the onset of gonadal steroidogenesis.

3-Hydroxysteroid Dehydrogenases↗
PubMed2001

Expression of steroidogenic factor-1 in frog embryo and developing gonad.

Steroidogenic factor-1 (SF-1), originally identified as an orphan nuclear receptor that regulates expression of genes encoding cytochrome P-450 steroid hydroxylases, is an essential transcriptional factor for adrenal and gonadal development in mammals. Since sex steroid hormones have been shown to play important roles in the sex determination of frogs, it is of interest to know how SF-1 gene expression is regulated during the sexual development of this organism. A previous study isolated the cDNA of the frog Rana rugosa SF-1 (rrSF-1) and found sexual differences in its gene expression in adult frogs; positive in testis and negative in ovary (Kawano et al., 1998). This study examined rrSF-1 gene expression in frog embryos and developing and mature gonads by immunohistochemical staining using anti-rrSF-1 protein antibody for protein localization and by in situ hybridization analysis for mRNA transcription. The results obtained in this study indicated that cells expressing SF-1 that originate in the mesoderm and endoderm probably migrate into the developing gonad via the dorsal mesentery, genital ridge, and mesorchium or mesovarium. Thus, SF-1 may play an important role in gonadal development in amphibians.

Adrenal Glands↗
PubMed1995

Arginine vasotocin injection increases probability of calling in cricket frogs, but causes call changes characteristic of less aggressive males.

Male cricket frogs, Acris crepitans communicate to males and females using advertisement calls, which are arranged into call groups. Calls at the middle and end, but not beginning of the call group, are modified in response to male-male aggressive interactions. We found in this field study of male cricket frogs in natural breeding choruses that the peptide hormone arginine vasotocin (AVT) not only increased the probability that males called after injections, but also caused modifications in middle and end calls to produce calls characteristic of less aggressive males. Moreover, AVT-injected males showed significantly greater increases in call dominant frequency than saline-injected males, again, a characteristic of less aggressive males. Cricket frog calls are used to both repel males and attract females, thus call changes may relate to male-male and/or male-female interactions. Saline-injected males also demonstrated significant changes in several call traits, including changes that occurred in the beginning and middle calls of the call groups, but not the end calls. AVT appeared to block some call changes produced through handling. These data suggest that AVT can influence acoustic communication in frogs in several ways, including effects on call characteristics and dominant frequency, as well as potentially blocking some handling effects.

Aggression↗
PubMed1990

Pharmacological modification of mechanical and electrical responses of frog heart to thrombin.

The pharmacological modification of the thrombin effect on the mechanical and electrical responses of frog heart was examined in the Straub heart preparation and in single ventricular cells. Associated with the positive inotropic action thrombin increases voltage and duration of action potentials of isolated frog ventricular cells. As found by the patch-clamp technique in the cell-attached mode, thrombin stimulates single L-type Ca2+ channels, presumably mediated by a second messenger. The enhancement of contractility by thrombin depends on the proteolytic activity of the enzyme because enzymatically inactivated thrombin has no effect on frog hearts. The positive inotropic effect of thrombin as well as its stimulation of Ca2+ channel currents were inhibited by the protein kinase C blocker 1-(5-isoquinoline-sulfonyl)-2-methylpiperazine (H7). However, phorbol 12-myristate 13-acetate (PMA), a known stimulator of protein kinase C, was ineffective in stimulating the inotropic action of thrombin. The inhibition of the thrombin-induced enhancement of contractility by indometacin indicates an involvement of arachidonic acid in the action of thrombin on frog heart.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1996

The roles of visual and proprioceptive information during motor program choice in frogs.

Previous studies have shown that leopard frogs, Rana pipiens, use tongue prehension to capture small prey and jaw prehension to capture large prey. After hypoglossal nerve transection, the frogs fail to open their mouths when attempting to feed on small prey, but open their mouths and capture large prey. Here, we investigate how visual information about the prey and proprioceptive information from the tongue interact to influence the motor program choice. Using pieces of earthworm of various sizes, we found that Rana exhibits two different behavior patterns based on prey size. The frogs captured the 1.5-cm prey using tongue prehension, whereas 2.0-cm and larger prey were captured using jaw prehension. After hypoglossal transection, the frogs never opened their mouths when they tried to feed on 1.5-cm prey. When feeding on 3.0-cm and larger prey after transection, they always opened their mouths and captured the prey using jaw prehension. When offered 2.0-cm prey, they alternated randomly between opening and not opening the mouth. Therefore, deafferentation changed the pattern of motor program choice at the behavioral border. This implies that afferents from the tongue interact with visual input to influence motor program choice.

Analysis of Variance↗
PubMed1996

The uptake and effects of lead in small mammals and frogs at a trap and skeet range.

This study was performed to evaluate the bioavailability and effects of lead in wildlife at a trap and skeet range. The total lead concentration in a composite soil sample (pellets removed) was 75,000 micrograms/g dry weight. Elevated tissue lead concentrations and depressed ALAD activities in small mammals and frogs indicate that some of the lead deposited at the site is bioavailable. Mean tissue lead concentrations (micrograms/g dry wt.) in white-footed mice (Peromyscus leucopus) at the range liver = 4.98, kidney = 34.9, femur = 245) were elevated (P < 0.01) 5- to 64-fold relative to concentrations in mice from a control area. Tissue lead concentrations in the only shorttail shrew (Blarina brevicauda) captured at the range (liver = 34.1, kidney = 1506, femur = 437) were elevated 35- to 1038-fold. Femur lead concentrations in green frogs (Rana clamitans) at the range (1,728 micrograms/g) were elevated nearly 1000-fold, and the lead concentration in a pooled kidney sample (96.2 micrograms/g) was elevated 67-fold. There was significant depression of blood ALAD activity in mice (P = 0.0384) and depression of blood and liver ALAD activity in frogs (P < 0.001). Hematological and histopathological lesions associated with lead toxicosis were observed in some animals. Hemoglobin concentrations were reduced 6.7% in mice (P = 0.0249), but hematocrit was not significantly affected in mice or frogs. Intranuclear inclusions were present in the renal proximal tubular epithelium of two of the mice and the shrew that were captured at the range, and necrosis of the tubular epithelium was also evident in one of the mice. Kidney:body weight ratios were similar in range and control mice. Soil ingestion may be a significant route of lead uptake in small mammals at the range. However, the tendency of lead to concentrate in the bones rather than in more digestible soft tissues may minimize food chain uptake of lead by predators, especially raptors that regurgitate undigestible material.

Animals↗
PubMed1991

Effects of serum, its protein and lipid extracts, and commercial serum proteins and lipid on the isolated frog heart.

This study investigates the inotropic effects of serum, its protein and lipid extracts, and commercial serum proteins and lipid on the isolated, spontaneously-beating heart and superfused, hypodynamic ventricle of the frog. Serum taken from either man, horse, calf, frog, or rabbit evoked marked positive inotropic responses which were unaffected by cholinergic, serotonergic, and adrenergic receptor antagonists. Dialysed serum (dialisand) and void volume fractions from Sephadex G200-120 columns corresponding to large molecular weight constituents evoked marked positive inotropic responses. When serum was separated into fractions containing either proteins or lipids/lipoproteins by high-density ultracentrifugation or activated charcoal, both extracts evoked marked positive inotropic responses. Commercial serum globulins and serum containing a high proportion of immunoglobulins elicited large increases in contractile force, whereas serum albumin evoked a negative inotropic effect. Serum which was either boiled and/or treated with chymotrypsin to denature proteins also caused a marked increase in isometric twitch tension in the frog heart. Similar inotropic response was obtained with fractions of boiled serum eluted on columns of Sephadex G200-120. These fractions corresponded to molecular weight in the region of 60-70 kDa. However, the inotropic effect of boiled serum was abolished following pretreatment with lipase. Superfusion of frog hearts with commercial cardiolipin resulted in marked dose-dependent increases in contractile force. The results demonstrate the presence of at least two large molecular weight cardioactive principles in serum. These substances are comparable in size to constituents of serum proteins (e.g., globulins and immunoglobulins) and serum lipids/lipoproteins (e.g., cardiolipin) and may serve as physiological regulators of cardiac function.

Animals↗
PubMed1995

Adaptations of the reed frog Hyperolius viridiflavus (Amphibia, Anura, Hyperoliidae) to its arid environment. VII. The heat budget of Hyperolius viridiflavus nitidulus and the evolution of an optimized body shape.

Estivating reed frogs of the superspecies Hyperolius viridiflavus are extraordinarily resistant to the highly adverse climatic conditions prevailing in their African savanna habitats during dry season (air temperature up to 45 degrees C, solar radiation load up to 1000 W.m-2, no water replenishment possible for up to 3 months). They are able to withstand such climatic stress at their exposed estivation sites on dry plants without evaporative cooling. We developed a heat budget model to understand the mechanisms of how an anuran can achieve this unique tolerance, and which allows us to predict the anuran's core and surface temperature for a given set of environmental parameters, to within 4% of the measured values. The model makes it possible to quantify some of the adaptive mechanisms for survival in semiarid habitats by comparing H. viridiflavus with anurans (H. tuberilinguis and Rana pipiens) of less stressful habitats. To minimize heat gain and maximize heat loss from the frog, the following points were important with regard to avoiding lethal heat stress during estivation: 1) solar heat load is reduced by an extraordinarily high skin reflectivity for solar radiation of up to 0.65 under laboratory and even higher in the field under dry season conditions. 2) The half-cylindrical body shape of H. viridiflavus seems to be optimized for estivation compared to the hemispheroidal shape usually found for anurans in moist habitats. A half-cylinder can be positioned relative to the sun so that large surface areas for conductive and convective heat loss are shielded by a small area exposed to direct solar radiation. 3) Another important contribution of body shape is a high body surface area to body mass ratio, as found in the estivating subadult H. viridiflavus (snout-vent lengths of 14-20 mm and body weights of 350-750 mg) compared to adult frogs (24-30 mm, 1000-2500 mg) which have never been observed to survive a dry season. 4) These mechanisms strongly couple core temperature to air temperature. The time constant of the core temperature is 29 +/- 10 s. Since air temperature can be 43-45 degrees C, H. viridiflavus must have a very unusual tolerance to transient core temperatures of 43-45 degrees C. 5) If air temperature rises above this lethal limit, the estivating frog would die despite all its optimizations, but moving from an unsuited to a more favorable site during estivation can be extremely costly in terms of unavoidably high evaporative water loss. Therefore, H. viridiflavus must have developed behavioral strategies for reliably choosing estivation sites with air temperature staying on average within the vital range during the whole dry season.

Acclimatization↗
PubMed1993

Metabolic depression and Na+/K+ gradients in the aestivating Australian goldfields frog, Neobatrachus wilsmorei.

During aestivation the metabolic rate of the Australian goldfields frog Neobatrachus wilsmorei was reduced by 80% from its standard metabolic rate. The in vitro rate of oxygen consumption of isolated muscle and skin from aestivating frogs was up to 50% lower than that of the non-aestivating frogs. This in vitro rate of oxygen consumption was maintained for 6-12 h, indicating an intrinsic metabolic depression of tissues during aestivation. Frogs became dehydrated during aestivation. Muscle, skin and liver also became dehydrated during aestivation, but brain and kidney did not. Na+ and K+ contents and extracellular space measurement for muscle indicated that ion gradients were maintained across the muscle cell membrane during aestivation. Increases in plasma concentrations of Na+ and K+ were matched with similar increases in muscle intracellular ion concentrations. Extracellular space measurements were unsuccessful in the other tissues, but K+ content in all tissues (per dry weight) was maintained during aestivation, and the concentration of plasma K+ did not increase above that which can be accounted for by dehydration, indicating that K+ gradients were maintained.

Animals↗
PubMed1990

Adenylate cyclase and carbonic anhydrase in the semicircular canal epithelium of the frog Rana esculenta. An ultrastructural cytochemical localization.

Because the secretion of endolymph has been localized in the ampullar part of the frog semicircular canal, we attempted to determine by cytochemical methods the ultrastructural localization of two enzymes that are assumed to play a role in endolymph secretion: carbonic anhydrase and adenylate cyclase. Functionally, the epithelium of the frog semicircular canal can be schematically divided into three areas: sensory (crista ampullaris), secretory (dark cells), and non-sensory and nonsecretory (transitional and undifferentiated cells) areas. Carbonic anhydrase activity was widely distributed in dark cells. Dark cell labeling disappeared in the presence of acetazolamide. The other cells of the canal did not show any carbonic anhydrase labeling except for the supporting cells of the sensory cells. Adenylate cyclase activity was found on the basolateral and apical membranes of dark cells, and on the apical membrane of sensory cells; weak labeling was also observed in the other epithelial cells. In the apical membrane of the dark cells, adenylate cyclase labeling was dependent on the presence of vasotocin, the frog antidiuretic hormone. The dark cells of the frog semicircular canal thus possess the enzyme equipment needed for the secretion of endolymph and its possible hormonal regulation.

Adenylyl Cyclases↗
PubMed1989

Changes in Na channel properties of frog and rat skeletal muscles induced by the AaH II toxin from the scorpion Androctonus australis.

The effects of the mammal toxin II isolated from the venom of the scorpion Androctonus australis Hector (AaH II) were studied under current and voltage clamp conditions in frog (semitendinosus) and rat (fast e.d.l. and slow soleus) skeletal twitch muscle fibres. In both species, AaH II induced a dose-dependent prolongation of the action potential (AP) leading at saturating concentration to APs with long plateaus of about 1.5 s in frog and 5 s in rat e.d.l. and soleus fibres. The concentrations to induce 50% of the maximal effect (K0.5) were 9.1 x 10(-9) M in the frog and 1.4 x 10(-9) M in the rat. AaH II increased the time constants of inactivation of the peak Na current and induced a maintained Na current that was greater in rat e.d.l. and soleus (31.6% of peak current amplitude at -30 mV; K0.5 = 0.8 x 10(-9) M) than in frog (16.5%; K0.5 = 15.5 x 10(-9) M) muscles. Peak and maintained Na currents were TTX-sensitive and had identical threshold and reversal potentials. The half-maximum maintained permeability occurred at a potential 20 mV more positive than the peak permeability. Recovery from inactivation and steady-state inactivation of the inactivating Na current remained unchanged. The maintained current deactivated with normal fast kinetics. The action of the toxin reversed poorly on washout but could be largely removed by conditioning depolarizations more positive than the reversal potential of the Na current. Our results suggest that, in vertebrate skeletal muscle fibres, AaH II affects all the Na channels and are consistent with the hypothesis that the maintained current originates from a reopening of previously inactivated Na channels.

Action Potentials↗