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PubMed1987

A sensitive in vitro toad skin bioassay for melanotropic peptides.

1. An in vitro bioassay for melanotropic peptide utilizing reflectance measurements of toad skin (Bufo ictericus ictericus) is described as an alternative to the commonly used Rana pipiens bioassay. 2. The toad skin bioassay is as sensitive to melanotropins and melanin concentrating hormone (MCH) as the frog bioassay. 3. On the basis of parallel dose-response curves obtained with the toad skin assay we found that beta-MSH is slightly less active than alpha-MSH, whereas the synthetic analogue [Nle4-D-Phe7]-alpha-MSH is about 10 times more potent and exhibits prolonged biological activity. 4. MCH, a putative neurohormone, can also be bioassayed in the in vitro toad skin bioassay, since it has alpha-MSH-like activity on amphibian melanocytes. 5. Since neither adreno- nor cholinoceptors are present in the toad melanocytes, the assay provides great specificity and sensitivity for the determination of melanotropin activity in tissue or blood.

Animals↗
PubMed1983

45Ca fluxes in isolated toad bladder epithelial cells: effects of agents which alter water or sodium transport.

Vasopressin enhances osmotic water flow and sodium transport across the toad urinary bladder by mechanisms involving cyclic AMP and calcium. It is believed that changes in intracellular calcium concentration or in its binding to membranes may in part mediate the effects of vasopressin. In addition, several agents which alter the response of the toad bladder to vasopressin may also act by altering cellular calcium metabolism. The effects of vasopressin and several agents which modify its effects in the toad bladder were studied on 45Ca fluxes in isolated epithelial cells from the toad bladder. Compartmental analysis of 45Ca exchange revealed three components. Vasopressin reduced the amount of calcium in the most rapidly exchanging pool from 1.67 +/- 0.20 to 0.86 +/- 0.12 nmol/mg of protein (P less than .025) and the most slowly exchanging pool from 2.72 +/- 0.26 to 1.90 +/- 0.34 nmol/mg of protein (P less than .001), while not affecting the intermediate pool. Theophylline, which mimics the natriferic and hydroosmotic effects of vasopressin, also mimicked the effects on 45Ca exchange by vasopressin. Exogenous cyclic AMP and the prostaglandin endoperoxide analog U46619, which mimic the hydroosmotic effect of vasopressin, also reduced the amount of calcium in the most slowly exchanging pool. Prostaglandin E1, which inhibits the hydroosmotic effect of vasopressin at the concentrations used increased the size of the most slowly exchanging pool. These studies suggest that prostaglandins and other agents which alter the effect of vasopressin in the isolated toad bladder may elicit their effects in part by influencing the calcium concentration at some critical site.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1981

Studies on the erythropoietic effect of testis of toad at a simulated high altitude (hypoxic exposure).

Sham operated control toads (non castrated) have shown increased activity in regard to the hematological parameters in comparison to the castrated toads kept under simulated hypoxic condition. Injection of plasma from sham operated control toads exposed to hypoxic stimulation initiates erythropoiesis in starved toads, whereas plasma from castrated exposed to the same hypoxic stimulation failed to do so. It can be suggested that in toad, testis is the only organ responsible for controlling erythropoiesis by secreting certain substance in the blood, which acts as erythropoietic stimulating factor (ESF), and the production of this factor was found to increase in response to the hypoxic stimulation in order to combat under low atmospheric condition.

Altitude↗
PubMed1978

[Glands in the skin of the fire-bellied toad (Bombina bombina(L.); Discoglossidae, Anura): type, number, size and distribution under natural and experimental conditions].

1. In the skin of the fire-bellied toad (Bombina bombina) 5 types of glands are found. By analogy with the comparable glands of the yellow-bellied toad and the grass frog, these are called the toxic, lumpy, mucous, callous, and small glands. The callous glands were omitted from the present study. 2. Lumpy glands are most numerous in the middle of the back of the fire-bellied toad, their numbers decreasing progressively over the sides to the middle of the ventral surface. Mucous glands are more common cranially than caudally. The toxic and small glands are uniformly distributed over the body. 3. On the average there are 700 glands per square cm of skin in the fire-bellied toad. Of these, in the male the lumpy glands represent less than 10%, while the toxic, mucous and small glands each account for 30%. In the females 50% of the total number of glands are mucous glands, and only about 10% toxic glands; lumpy and small glands occur in the same proportions as in the males. Following gonadotropin injection, no change in the number of glands was observed in the male. 4. The largest toxic and lumpy glands are located cranially; toward the tail their size is reduced. The mucous glands have the greatest volume on the sides of the body. Small glands are of similar size everywhere. 5. Glands of uniform size are arranged in longitudinal rows on this toad. The dorsally situated toxic, lumpy and mucous glands are larger, on the whole, than the more ventral glands of the same types. 6. All 4 gland types on the body of Bombina are larger in males than in females. 7. Following administration of PMS to the male there is a selective increase in volume of the toxic and lumpy glands on the dorsal surface, but no corresponding increase on the ventral surface. Mucous and small glands do not change in size after hormone treatment. 8. The capacity of all the glands in a cm2 of skin, in terms of mean volume, is 1.7 mm3 in males not treated with hormones. After PMS treatment of the male the gland volume per cm2 increase to 2.5 mm3. The total volume per cm2 in females is only 0.65 mm3.

Animals↗
PubMed1993

[Type and content of adrenoceptors in toad heart].

The type and content of adrenoceptors in heart of normothermic and cold-acclimatized toad were studied by radioligand binding assay at different testing temperatures. The Bmax and Kd values of membrane from normal toad heart binding to 3H-DHA at 37 degrees C were respectively 55.11 +/- 6.22 fmol/mg protein and 2.15 +/- 0.42 nmol/L for the whole heart, 55.80 +/- 7.03 and 2.65 +/- 0.37 for sino-atrium and 54.27 +/- 3.06 and 1.84 +/- 0.14 for ventricle. While the binding to 3H-DHE was very low and nonspecific. When membrane from cold-acclimatized toad at 5-8 degrees C for 10 days was examined for its binding to 3H-DHA or 3H-DHE at 10 degrees C, the values of Bmax and Kd were as same as those of normothermic toad examined at 37 degrees C. The present results suggest that the toad heart is lacking of alpha-adrenoceptor, and the type and content of adrenoceptors are not influenced by temperature.

Animals↗
PubMed1997

Effects of dry season dormancy on oxygen uptake, heart rate, and blood pressures in the toad, Bufo paracnemis.

The cardiodynamic consequences of dry season dormancy in ectothermic vertebrates is not well known. Our hypothesis was that dormancy would reduce cardiac activity. We therefore determined oxygen uptake and cardiovascular function in aestivating toads, Bufo paracnemis, native to São Paulo State, Brazil. Specimens were collected and kept in the laboratory under controlled temperature and light regimes. We compared oxygen uptake, heart rate, blood pressure, rate-pressure product (RPP), and blood gases in toads during aestivation (dry winter season) and their early active season (spring). Oxygen uptake of winter toads at 25 degrees C was considerably lower than that of spring toads (winter: 24.0 +/- 1.8 ml/(kgh); early spring: 44.4 +/- 5.1 ml/(kgh); mean +/- SE; same in the following). A seasonal dichotomy was also observed at 15 degrees C although the differences was less pronounced (15.8 +/- 1.8 ml/(kgh) winter; 23 +/- 2.1 ml/(kgh) early spring). Chronic arterial cannulation permitted measurements of cardiodynamic variables without any undesired change in VO2. Heart rates of winter toads were significantly lower than those of early spring animals at both experimental temperatures (25 degrees C: winter 25 +/- 1.4 beats/min.; early spring: 35.2 +/- 5.1 beats/min. 15 degrees C: winter 15, 4 +/- 1.8 beats/min.; early spring: 23.9 +/- 2.1 beats/min). Systemic, diastolic and mean arterial pressures decreased slightly but not significantly during aestivation. We conclude that: (1) Bufo paracnemis downregulates metabolic rate during the dry season and (2) heart rate is also downregulated with little change of blood pressure. While the energetics of these responses are probably beneficial for survival during aestivation, the underlying biochemical mechanisms remain obscure.

Animals↗
PubMed1996

[Seasonal variation and circannual rhythm of miniature end-plate potentials recorded from the neuromuscular junctions of the adult toad (Bufo bufo gargarizans)].

The seasonal variations and circannual rhythm of miniature end-plate potentials (MEPP) recorded from the sartorius neuromuscular junctions (NMJ) of 66 female and 62 male adult toads (Bufo bufo gargarizans) were studied with intracellular microelectrode recording techniques of electrophysiology. The results revealed that the mean amplitude of MEPP in each month of a year showed clear seasonal variation with a nadir in September and October and a zenith in March and April. This seasonal variation in MEPP amplitude was proved to be a circannual rhythm by single cosinor analysis and zero amplitude test, and the three parameters of the rhythm, i.e. mesor, amplitude and acrophase were 0.29 +/- 0.05 mV, 0.25 mV and -97.17 degrees in female toad and 0.19 +/- 0.02 mV, 0.16 mV and -92.01 degrees in male toad, respectively; the circannual rhythm in female toad was more distinct than that in male. Neither seasonal variation nor circannual rhythm was statistically significant in the changes of frequency of MEPP, regardless of toad sexuality.

Animals↗
PubMed1999

Desert toads discriminate salt taste with chemosensory function of the ventral skin.

Toads obtain water by absorption across their skin. When dehydrated, desert toads exhibit stereotyped hydration behavior in which they press their ventral skin onto a moist surface. However, dehydrated toads avoid surfaces moistened with hyperosmotic NaCl and KCl solutions (Hoff KvS, Hillyard SD. 1993. J. Exp. Biol. 183:347-351). We have studied neural mechanisms for this avoidance with physiologic, behavioral, and morphologic approaches. Spinal nerves innervating the ventral skin could be stimulated by exposure to a hyperosmotic NaCl solution applied to the outer surface of the skin. This neural response occurred with much longer latency than to mechanical stimulation and could be reduced by amiloride, a blocker for Na+ channels known to be responsible for epithelial ion transport and salt taste transduction. In behavioral experiments, avoidance of a NaCl solution was also reduced by adding amiloride to the solution, suggesting involvement of amiloride-sensitive Na+ channels for detecting the hyperosmotic salt solution. Neural tracing with fluorescent dye revealed spinal nerve endings and connections to putative receptor cells, both located in the deeper layer of the epidermis. Either of these or both may be associated with the transduction of Na+ flowing into the skin. The ability of toads to detect hyperosmotic salt solutions in their environment reveals a previously unknown chemosensory function for spinal nerves in anuran amphibians.

Action Potentials↗
PubMed1999

Skull allometry in the marine toad, Bufo marinus.

Scaling predictions pioneered by A.V. Hill state that isometric changes in kinematics result from isometric changes in size. These predictions have been difficult to support because few animals display truly isometric growth. An exception to this rule is said to be the toads in the genus Bufo, which can grow over three orders of magnitude. To determine whether skull shape increases isometrically, I used linear measurements and geometric morphometrics to quantify shape variation in a size series of 69 skulls from the marine toad, B. marinus. Toads ranged in body mass from 1.8 gm to a calculated 1, 558.9 gm. Of all linear measurements (S/V length, skull width, skull length, levator mass, depressor mass, adductor foramen area), only the area of the adductor foramen increased faster than body mass; the remaining variables increased more slowly. In addition, modeling the lower jaw as a lever-arm system showed that the lengths of the closing in- and out-levers scaled isometrically with body mass despite the fact that the skull itself is changing allometrically. Geometric morphometrics discerned areas of greatest variability with increasing body mass at the rear of the skull in the area of the squamosal bone and the adductor foramen. This increase in area of the adductor foramen may allow more muscle to move the relatively greater mass of the lower jaw in larger toads, although adductor mass scales with body mass. If B. marinus feeds in a similar manner to other Bufo, these results imply that morphological allometry may still result in kinematic isometry.

Animals↗
PubMed1984

Uptake of 3H-glycine in the outer plexiform layer of the retina of the toad, Bufo marinus.

The uptake of 3H-glycine in the retina of the toad, Bufo marinus, was investigated by light and electron microscopical autoradiography. Uptake of 3H-glycine was very prominent in large cell bodies in the inner nuclear layer as well as in discrete clusters in both the outer plexiform layer (OPL) and the inner plexiform layer. This pattern in similar to that described for 3H-glycine-accumulating putative interplexiform cells in goldfish, frog, and Xenopus retinas. Electron microscopical autoradiography of the OPL revealed large, grain-containing varicosities which had electron-lucent cytoplasm and contained both small, agranular and large, dense-core vesicles. The varicosities made extensive en passant and spine synapses in the OPL. Definitive identification of their postsynaptic targets was not achieved. However, autoradiographic analysis with 3H-GABA uptake as well as electrophysiological evidence suggests that axons but not cell bodies or dendrites of 3H-GABA-accumulating horizontal cells (H1 cells) are postsynaptic targets of the varicosities. The presence of dense-core vesicles in the varicosities suggested co-occurrence of glycine and a biogenic amine or neuropeptide. The indirect immunofluorescence technique was used to determine whether any such substances were present in the OPL of the toad retina. However, no specific labeling was found in the OPL for any of 19 substances tested. The extensive synaptic output provided by glycine-accumulating varicosities in the toad OPL may indicate an important role of glycine in the synaptic function of the distal toad retina. We suggest that these varicosities derive from a presumably glycinergic interplexiform cell.

Animals↗
PubMed1995

Early postmitotic neurons transiently express TOAD-64, a neural specific protein.

To identify proteins involved in the early development of the mammalian cerebral cortex, we previously used two-dimensional gels to compare proteins synthesized at different stages in corticogenesis in the embryonic rat at embryonic day 14 (E14), E17, and E21. During this period, the cortex develops from a morphologically homogeneous population of proliferative precursor cells into a complex structure containing a diverse array of terminally differentiated neurons. Several proteins are up-regulated coincident with the generation of postmitotic neurons. Here we describe the purification, partial amino acid sequencing, and characterization of one of these proteins, TOAD-64 (Turned On After Division; 64 kDa), using polyclonal antisera to two synthetic peptides from the protein. This analysis reveals that TOAD-64 is a 64,000 Da protein that increases in abundance over the period of corticogenesis and then subsequently decreases to very low levels in the adult. The protein is neural specific and is expressed by postmitotic neurons as they begin their migration out of the ventricular zone into the developing cortical plate. It is expressed in advance of most other neuronal proteins. Progenitor cells do not express TOAD-64. Therefore, this protein is a marker for postmitotic cells that have made a commitment to a neuronal phenotype. The extremely early expression, the relative abundance in newly born neurons, as well as the restriction in expression to the period of initial neuronal differentiation suggest that TOAD-64 may be a key structural protein for early neuronal function.

Amino Acid Sequence↗
PubMed1982

Factors controlling the ovarian cycle in a temperate zone anuran, the toad Bufo bufo: food uptake, nutritional state, and gonadotropin.

The relations between nutritional state and vitellogenic growth of oocytes were studied in 87 sexually mature female toads Bufo bufo during the postbreeding season before initiation of the next annual ovarian cycle. Recruitment of small oocytes to vitellogenic growth was correlated with resumption of food uptake and the restoration of a positive energy balance in the toads that had become depleted of energy reserves during hibernation and breeding. Treatment with exogenous gonadotropin (hCG) immediately initiated vitellogenic growth, independently of the nutritional state. But the hCG-induced vitellogenic growth was faster in toads that ate more than the maintenance ration than in those that ate less. hCG also maintained rapid vitellogenic growth in toads that starved after an initial period of feeding during which vitellogenic growth had been resumed. The results support the hypothesis that the period of ovarian quiescence, which normally follows breeding in Bufo bufo, and other anurans from the northern range of distribution of the species, results from inactivation of the hypothalamic-hypophysial gonadotropic system, and that reactivation of the system depends upon reestablishment of a proper nutritional state of the organism.

Animal Nutritional Physiological Phenomena↗
PubMed2004

Vasculature of the parotoid glands of four species of toads (bufonidae: bufo).

The parotoid glands of toads (Bufonidae) consist of large aggregations of granular glands located between the otic region of the skull and the scapular region. To determine the circulatory pattern of these glands, we perfused the vascular systems of Bufo alvarius, B. marinus, B. terrestris, and B. valliceps with either India ink or Microfil, a fine latex. The perfused glands were studied by gross dissection, microscopic examination, and histology. The vascular patterns of the parotoid glands were compared to the arrangement of vessels in the dorsal skin of Rana sphenocephala (Ranidae), a frog that lacks parotoid glands. The parotoid glands of the four species of toads are supplied with blood by the lateral and dorsal cutaneous arteries and are drained by one or more branches of the internal jugular vein. The dorsal cutaneous artery supplies most of the blood to the parotoid glands in B. terrestris and B. valliceps. In B. alvarius and B. marinus, both the lateral and dorsal cutaneous arteries serve major roles in the blood supply of the glands. These patterns of blood flow have not been described previously for parotoid glands and conflict with earlier accounts for B. alvarius and B. marinus. The arteries and veins associated with the parotoid glands of toads are present in R. sphenocephala, but are arranged differently. In R. sphenocephala, the lateral cutaneous artery supplies the dorsal and lateral skin posterior to the shoulder region, whereas the dorsal cutaneous artery supplies the skin of the shoulder region. In toads, both the lateral and dorsal cutaneous arteries supply the skin of the shoulder region and ramify into subcutaneous capillaries that surround the secretory units of the parotoid glands. Extensive vasculature presumably is important for delivering cholesterol and other precursor molecules to the parotoid glands, where those compounds are converted into toxins.

Animals↗
PubMed1993

Metabolism and cardiovascular effects of leukotrienes in the marine toad Bufo marinus.

Leukotriene (LT) metabolism and physiology have been studied extensively in mammals; however, little is known of their roles in nonmammalian vertebrates. This study examines the cardiovascular effects of leukotrienes on blood pressure and heart rate in the conscious and cannulated marine toad, Bufo marinus. The sulfidopeptide leukotrienes, LTC4, LTD4, and LTE4 elicited hypotension with equal potency. However, with respect to heart rate changes and duration of action, the responses to LTC4 and LTD4 were greater and lasted longer than those to LTE4. The nonpeptide leukotriene, LTB4, had significantly less potent effects on heart rate and blood pressure. The leukotriene-induced increases in heart rate with 1000 and 300 ng/kg body wt LTC4 and LTD4 were blocked with 5 mg/kg body wt propranolol, a beta-antagonist, suggesting sympathetic reflex regulation of heart rate. Metabolism of [3H]LTC4 to [3H]LTD4 and [3H]LTE4 occurred rapidly in blood, with complete conversion to [3H]LTE4 within 5 min. Conversion was slower in plasma, with 18.9 +/- 0.5% of the radioactivity associated with [3H]LTC4 still remaining after 120 min. The toad is more similar to mammals than the bullfrog with respect to the metabolism of leukotrienes. In contrast to mammals, leukotrienes have hypotensive effects in both toad and bullfrog, although the order of potency differs. The effectiveness of the sulfidopeptide leukotrienes in eliciting hypotension at low doses (1 ng/kg body wt) suggests that these compounds may be important cardiovascular regulators in the toad.

Analysis of Variance↗
PubMed1994

Metabolism of toad ventricle during alterations to the inotropic state.

The heat produced by toad ventricle during manipulations of the inotropic state was measured using thermopiles, and some comparisons made to rat ventricle. The tension-independent heat, peak stress, and the tension-dependent heat increased when [Ca2+]o increased from 0.25 to 2 mM in Ringer. In 2 mM [Ca2+]o, tension-independent heat, peak stress, and tension-dependent heat were 3.1 +/- 0.4 mJ/g, 38.4 +/- 5.5 mN/mm2, and 0.49 +/- 0.06 units; about 25% of the tension-independent heat may relate to the Na(+)-K+ pump. At similar [Ca2+]o, rat ventricle produced a smaller tension-independent heat (1.6 +/- 0.2 mJ/g), and active heat per unit stress (0.22 +/- 0.01 units) than toad. Tension-independent heat, stress, and tension-dependent heat were increased by orciprenaline, and decreased by BDM. Ouabain increased the stress and tension-dependent heat but not the tension-independent heat. Five millimolar [Ca2+]o in HEPES buffer decreased the stress but increased the tension-dependent heat compared to 2 mM [Ca2+]o in Ringer. Ryanodine and CPA caused major reductions in force and tension-independent heat in rat, but had little effect on toad ventricle. In conclusion, our results suggest that in toad ventricle (a) the sarcoplasmic reticulum plays only a minor role in activation and relaxation, (b) the Na(+)-K+ pump contributes substantially to activation metabolism, (c) active metabolism is stimulated by increases in [Ca2+]o and (d) there is a larger tension-independent heat, a larger active metabolism per unit stress, and a lower basal metabolism than in rat papillary muscle. The energy cost of removing intracellular Ca2+ through the sarcolemma appears to be greater than uptake into sarcoplasmic reticulum.

Animals↗
PubMed1991

Effects of environmental conditions on mitochondrial-rich cell density and chloride transport in toad skin.

Chloride flux across amphibian skin is usually passive, yet largely conductive; previous reports have suggested that aldosterone influences this pathway. The conductive Cl- pathway and its regulation were examined further, across the abdominal skin of toads (Bufo marinus) adapted to various environments. Short-circuit current (Isc), total conductance (Gt) and Cl- influx (JCl) were measured in conditions such that there was net Cl- movement in absence of Na+ transport. In salt-deprived animals compared to salt-adapted ones, there was a significant increase in JCl (563 vs 200 pmol cm-2 s-1), aldosteronaemia (4.2 vs 1.1 nmol/l), as well as MRC density (1458 vs 851 mm-2). After adaptation to dilute Na2SO4 compared to MgCl2, JCl (631 vs 313 pmol cm-2 s-1) as well as the density of mitochondria-rich cells (MRC) (1306 vs 710 mm-2) practically doubled, while the toads' aldosteronaemia was lower (2.4 vs 10.8 mmol/l). In all groups of toads, JCl was matched by Isc, and there was a close correlation between Gt and JCl (r = 0.96), which confirms the conductive nature of transepithelial Cl- movement. Furthermore, the relationship between JCl and MRC density (r = 0.75) argues in favour of a role played by MRC on Cl- conductance of epithelial such as amphibian skin. As aldosterone injected for 1 week into NaCl-adapted toads did not influence MRC density and as aldosteronaemia was not correlated with Cl- conductance, this hormone does not emerge as the determinant of these parameters.

Aldosterone↗
PubMed1996

Effects of angiotensin II antagonists on the contractile and hydrosmotic effect of AT II and AT III in the toad (Bufo arenarum).

The effects of peptide and non-peptide angiotensin II receptor antagonists on the responses to angiotensin II were examined using aortic rings and skin isolated from the toad. The contractile responses of aortic rings to (Ala-Pro-Gly) angiotensin II were inhibited by angiotensin II analogue Leu8 angiotensin II, with a pA2 value of 7.6. Similarly, the concentration response curve for (Ala-Pro-Gly) angiotensin II was displaced to the right by the specific angiotensin receptor subtype antagonist DuP 753, with a pA2 value of 6.0. In contrast, the angiotensin receptor subtype 2 antagonists PD 123177 and CGP 42112A did not modify the contractile response to (Ala-Pro-Gly) angiotensin II. None of the antagonists was able to alter the contractile response to norepinephrine. Both Leu8 angiotensin II (10(-8) mol.1(-1)) and DuP 753 (10(-6) mol.1(-1)) partially inhibited angiotensin III-induced contractions in toad aorta. Angiotensin III, in turn, exhibited lower activity than [Asn1-Val5] angiotensin II in this preparation, its molar potency ratio being 0.293. Previous work from this laboratory reported that osmotic water permeability in the skin of the toad Bufo arenarum was increased by angiotensin II, the effect being blocked by the peptide antagonist Leu8 angiotensin II. The hydrosmotic response to [Asn1-Val5] angiotensin II(10(-7) mol.1(-1)) was significantly inhibited by DuP 753 (10(-6) and 5 x 10(-6) mol.1(-1)), whereas the response was not inhibited by a tenfold higher concentration of either PD 123177 or CGP 42112A. DuP 753 (10(-6) mol.1(-1) also inhibited the hydrosmotic response to angiotensin III (10(-7) mol.1(-1)). These results suggest that receptors for angiotensin II present isolated toad aorta and skin exhibit pharmacological features similar to those characterized as angiotensin subtype 1 in mammalian tissues.

Analysis of Variance↗
PubMed1981

Resistance of adrenergic neurotransmission in the toad heart to adrenoceptor blockade.

Stimulation of sympathetic nerves to the toad heart produced increases in both the rate and force of cardiac beat. Although these responses were abolished by treatment with bretylium (10-6 mol.1-1) or 6-hydroxydopamine (100 mg.kg-1), and surgical sympathetic denervation, they were not abolished by treatment with propranolol (10-6 mol-1-1) and phentolamine (3 X 10-6 mol.1-1), either alone or in combination. The responses remaining after adrenoceptor blockade could not be ascribed to the effects of neurally released dopamine, ATP, adenosine, histamine or a variety of neuropeptides, although the participation of an as yet unidentified co-transmitter cannot be ruled out. Quantitative analysis of the interactions between propranolol and adrenaline on cardiac adrenoceptors, after blockade of alpha-receptors and amine uptake mechanisms, revealed that these interactions do not comply with the conditions for simple competitivity. Therefore, in addition to its action on beta-receptors, adrenaline seems to be producing excitation of the toad heart by acting on adrenoceptors which cannot be classified as either alpha- or beta-receptors. These results, together with the existence of close neuromuscular gaps (less than 50 nm) in the toad heart, are consistent with the hypothesis that sympathetic excitation of the toad heart is mediated by both "extra-junctional" beta-adrenoceptors, and "junctional" adrenoceptors which are neither alpha- nor beta-receptors.

Animals↗