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A Grosso

Publications and source records attributed to A Grosso.

At least 37 records · Page 2Linked to original sources

High [K+] alters the stimulus-hydrosmotic response coupling in toad bladder.

Substitution of K+ for Na+ in the Ringer solution bathing the inner surface of toad urinary bladders (Bufo marinus) had no effect on basal water permeability but significantly altered the stimulus-hydrosmotic response of this epithelium. In chloride-Ringer, high [K+] increased the hydrosmotic responses to submaximal stimulations induced by vasopressin or exogenous cAMP, while the responses to theophylline or serosal hypertonicity were decreased. In sulfate-Ringer, all these responses were enhanced but for that induced by serosal hypertonicity which was actually diminished. As a step towards determining if Ca2+ might mediate the K+-induced effects on water flow, experiments were conducted either in the presence of a Ca2+ "antagonist" (cobalt) or in nominally Ca2+-free Ringer. In both conditions the hydrosmotic effects of vasopressin and cAMP were markedly reduced. The results raise the possibility that a transient Ca2+ influx through voltage-sensitive, Co2+-blockade Ca2+ channels may play a role in the stimulus-hydrosmotic response of toad urinary bladder.

Animals↗

Flavonoids and hepatic cyclic monophosphates in liver injury.

Among the large spectrum of pharmacological activities of flavonoids, play an important role the recently investigated properties involving the arachidonic acid metabolism. In order to clarify the mechanisms of "cytoprotection" of the 3-palmitoyl-(+)-catechin (Palm-cat), a new flavonoid compound (C31 H44 O7) we have studied in experimental hepatitis of the rat, induced by Galactosamine (Ga1N) and E. coli 055:B 5 endotoxin (LPS), hepatic cAMP and cGMP, transaminases, bilirubin and endotoxemia. The Palm-cat significantly increases cyclic-GMP levels in the liver, whereas reduces or slightly modifies the cAMP. Transaminases and bilirubin values increase both in controls and flavonoid treated rats. The flavonoid significantly decreases the frequency of endotoxemia. These effects suggest that RES and hepatocytes functions, immune and inflammatory response can be affected in liver disease by flavonoids via cyclic nucleotides regulation.

Animals↗

Correlation between water flow and intramembrane particle aggregates in toad epidermis.

In abdominal skins of toads (Bufo marinus) challenged with either vasopressin or isoproterenol, the stimulation of transepithelial water flow was accompanied by the appearance of intramembrane particle (IMP) aggregates. These aggregates were found only in the apical plasma membrane of granular cells of the first-reacting cell layer. The effect of vasopressin varied in magnitude and could be selectively blocked by preincubation of the skins with methohexital. This barbiturate also prevented the appearance of IMP aggregates. Preexposure with propranolol had a similar effect in skins challenged with isoproterenol. Linear regression analysis of net water flow vs. the percentage surface occupied by IMP aggregates showed a significant (P less than 0.001) correlation, which applied to data obtained in six different experimental conditions, i.e., vasopressin or isoproterenol alone and either of these agents in combination with methohexital or propranolol. The results reported here in a nonurinary epithelium (toad epidermis) suggest that the appearance of IMP aggregates is a general response in epithelia undergoing adenosine 3',5'-cyclic monophosphate-mediated changes in water permeability.

Animals↗

Cellular and membrane events involved in the K-induced increase in water permeability of toad skin.

Exposure of the inner surface of toad skin (Bufo marinus) to high [K+] resulted in a marked (up to 7-fold) increase in water permeability (Pf) that was more marked in KC1-Ringer than in K2SO4-Ringer. Although high [K+] did not elicit a maximal increase in Pf, it blunted the hydrosmotic responses to vasopressin, isoproterenol and cAMP. Both "post-cAMP" inhibitors of stimulated water flow, such as diamide and vanadate, and "pre-cAMP" inhibitors, such as methohexital and propranolol, markedly reduced the K response, while exposure to Ca2+-free, KC1-Ringer did not inhibit water flow. Intramembrane particle aggregates, similar to those induced by cAMP-mediated hydrosmotic agents, were seen in the apical membrane of granular cells, just beneath the stratum corneum, in skins exposed to KC1. Available evidence indicates that cAMP might mediate, at least partially, the hydrosmotic effect of high [K+]. In contrast, a role of voltage-dependent Ca2+ channels, described in other cell systems depolarized with K, was not apparent in toad skin.

Animals↗

Osmotic water flow across the abdominal skin of the toad bufo marinus: effect of vasopressin and isoprenaline.

1. Net water flow J(w), was measured across the abdominal skin of the toad Bufo marinus with a volumetric, automatic technique that allows for averaging J(w) over time intervals as short as 1 sec.2. Basal J(w) was very stable and corresponded to a coefficient of osmotic flow, L(PD), of ca. 15 x 10(-7) cm sec(-1) atm(-1) (or to an osmotic water permeability coefficient, P(f), of 20 mum sec(-1)).3. Both vasopressin and the beta-adrenergic agonist, isoprenaline, triggered high hydrosmotic responses that could lead to P(f) values exceeding 250 mum sec(-1). The effect of isoprenaline was very reproducible while that of vasopressin varied considerably.4. Methohexital and propranolol selectively inhibited the hydrosmotic effects of vasopressin and isoprenaline, respectively, whereas amiloride and ouabain had no effect.5. Mutual inhibition was found between vasopressin and isoprenaline in skins very sensitive to vasopressin. In less sensitive skins isoprenaline further increased J(w) despite exposure of the epithelia to supramaximal concentrations of vasopressin.6. Differential reactivity to vasopressin was found between the skin and the bladder taken from the same toad. In some instances, the bladder responded normally to vasopressin while the skin was totally unresponsive, suggesting the presence of osmoregulatory mechanisms exerting a local modulation of the vasopressin action in different target epithelia of the same animal.

Animals↗

Evidence for a role of calmodulin in the hydrosmotic action of vasopressin in toad bladder.

1. The informational role of cytosolic Ca2+ appears to be mediated by a ubiquitous protein--calmodulin--in most cell systems. 2. Evidence has been accumulating that not only cAMP, but also Ca2+, behaves as an intracellular messenger in the stimulation of water transport by vasopressin (hydrosmotic effect). 3. To examine whether calmodulin plays a role in the hydrosmotic effect of vasopressin, we used a specific antagonist of calmodulin--trifluoperazine--and looked at its effects on water transport in the urinary bladder of toads Bufo marinus. 4. The results showed that trifluoperazine, at micromolar concentrations, blocked the hydrosmotic effects of vasopressin or cAMP, thus indicating a post-cAMP site of action. 5. Two other psychotropic drugs--amitriptyline and harmaline--had similar effects, but higher concentrations were required to induce the same degree of inhibition of water flow. 6. Calmodulin was detected in the membrane and in the cytosolic fractions of isolated epithelial cells of toad bladder by means of the phosphodiesterase test. The content of both fractions was similar to that found in bovine brain. 7. The results suggest that calmodulin plays a regulatory role in the hydrosmotic action of vasopressin by possibly interacting with proteins associated with microfilaments and/or microtubules.

Amitriptyline↗

Quercetin enhances water transport in toad bladder.

A highly significant enhancement of the hydrosmotic actions both of vasopressin and of exogenous cAMP was seen in the presence of quercetin. The hypothesis is advanced that quercetin affects the intracellular coupling between Ca++ in cAMP.

Animals↗

The amphibian epidermis: distribution of mitochondria-rich cells and the effect of oxytocin.

It is known that the ion-transporting capacity and the permeability to water of amphibian skins vary greatly both between and within species. Furthermore, the extent to which different skins respond to hormonal stimulation of these parameters also shows considerable inter- and intra-specific variation. As a first step towards defining a possible morphological basis for this physiological heterogeneity, we examined different regions of skins from 3 anurans, Bufo bufo, Rana ridibunda and Xenopus laevis, that are species with widely differing habitats. The mitochondria-rich cell population of the epidermis was counted and the epidermal thickness was measured. There were large differences in the mitochondria-rich cell content and in the epidermal thickness of the skins from different species and from different regions of skin from the same animal. In a second set of studies, the same morphological features were examined and, in addition, routine functional parameters were measured to monitor some transport properties of the skins used. The skins also varied considerably with respect to short-circuit current, potential difference, water permeability and sensitivity to oxytocin. Although no apparent relationship was noted between either basal or hormone-stimulated physiological parameters and the morphological features of the individual skins, the striking variation in the density of mitochondria-rich cells in amphibian epidermis merits further studies, including the use of techniques or experimental designs that allow the movement of individual species of ion across the skin to be followed.

Animals↗

The mode of action of vasopressin: membrane microstructure and biological transport.

Vasopressin affects a variety of cell systems. This review is focused on permeability changes induced by vasopressin in tight epithelia such as the collecting duct of the mammalian kidney and the skin and the bladder of anurans. These vasopressin effects are discussed with reference to current concepts and models of the microstructure of the plasma membrane. The transport of three major chemical species--Na, urea and water--is analyzed. In each instance, the hormone appears to activate selective membrane pathways situated at the rat-limiting barrier of the epithelium, i.e., the apical membrane. Available data suggest that two intra-cellular messengers -- cAMP and calcium -- plan a key role in the coupling between stimulus (receptor occupancy) and biological effect (permeability change). The enhancement of Na transport (natriferic effect) depends on the opening and/or the insertion of Na channels, the biophysical and biochemical characteristics of which have been investigated by fluctuation analysis and by means of several chemical blockers of Na transport, particularly the amiloride molecule and its congeners. Likewise, the finding of inhibitors and activators of urea transport, which do not cause any appreciable change in Na or water permeability, led to the notion of selective urea channels or pores. Finally, the enhancement of water transport (hydrosmotic effect) possibly results from the insertion in the apical membrane of water channels already present in vesicular cytoplasmic structures. The restructuring of the apical membrane underlying the transition from a low to a higher state of water permeability is very likely related to the appearance of intramembrane particle aggregates detectable with the freeze-fracture technique in epithelia exposed to vasopressin. The putative water channels (or pores) appear to be so narrow that trans-apical water movement is constrained to single-file diffusion. Recent data also suggest that, in addition to cAMP, microtubules and microfilaments, the calmodulin-Ca complex is a major element in the hydrosmotic effect of vasopressin.

Amphibians↗

Isoproterenol-induced intramembrane particle aggregation and water flux in toad epidermis.

Stimulation of toad skin with isproterenol resulted in a dramatic increase in water flow, and in the appearance of aggregates of intramembrane particles in the apical membrane of granular cells of the replacement layer, just beneath the stratum corneum. This membrane structural modification appears to be a general prerequisite for the change in water permeability of vasopressin-sensitive epithelia.

Animals↗

Cytochalasin B and water transport. A scanning electron microscope study of the toad urinary bladder.

A morpho-functional study of the effects of cytochalasin B (CB) on Na and water transport was made in amphibian epithelia. The functional studies confirmed the dissociation of the natriferic and hydrosmotic effects of vasopressin in toad urinary bladders exposed to CB and showed in addition that the block of the hydrosmotic effect was reversible and could still be induced in epithelia maximally stimulated with the hormone. Scanning electron microscopy revealed that CB, per se, did not alter the apical surface of the bladders. An almost total loss of microvilli of granular cells was seen, however, if CB was associated with vasopressin and an osmotic gradient. The results suggest two points: a) the block of the hydrosmotic flow induced by CB is due to factors beyond the apical membrane; b) microfilaments may be important mechanochemical transducers in the chain of events leading to the hydrosmotic effect of vasopressin.

Animals↗

Vasopressin-like effects of a hallucinogenic drug--harmaline--on sodium and water transport.

To determine if harmala alkaloids affect transport systems other than (Na +K)-ATPase, effects of harmaline on Na and water fluxes were studied in amphibian skins. Net Na flux was evaluated from short-circuit current, and water flux monitored with automatic, volumetric methods. At 2 to 5 mM, harmaline consistently inhibited SCC and prevented the natriferic effects of oxytocin and norepinephrine. However, at 0.1 to 0.5 mM, harmaline produced an increase in SCC inhibitable with amiloride. The stimulatory effects of harmaline and oxytocin were either nonadditive or additive depending on whether the hallucinogen was present in the inner solution or in the outer solution bathing the skin, respectively. Water flow was not modified by harmaline on the outer medium. In contrast, addition of the drug to the inner medium elicited a conspicuous, sustained, vasopressin-like, hydrosmotic effect, comparable to and competive with those of vasopressin and norepinephrine. The ensemble of these results suggests that harmaline may affect three distinct transport systems: (i) the Na pump; (ii) the cyclic nucleotide system; (iii) the Na entry pathway at the outer membrane of the skin that is also activated by agents such as diphenylhydantoin, lanthanides and propranolol.

Alkaloids↗

Vasopressin-like effects of psychotropic drugs in amphibian epithelia.

Amphibian epithelia have been used as models for studying the effects of psychotropic drugs on membrane transport. Several of these agents added to the internal or to the external media, at concentrations greater than 10(-3) M, had inhibitory, "ouabain-like" effects on Na transport. In contrast, stimulatory, "vasopressin-like" effects were seen at lower concentrations. The stimulation was additive to that of oxytocin if the drug was present in the external solution but nonadditive if in the internal solution. On water transport, harmala alkaloids had a vasopressinomimetic action in toad skin, while inhibition was seen with Li and amitriptyline. To account for these multiple effects, it is hypothesized that psychotropic drugs act on the following cell targets: the Na pump, the cyclic nucleotide system, microtubules, and membrane calcium sites at the outer barrier of the epithelium. Direct, biochemical evidence is needed to substantiate this hypothesis.

Amitriptyline↗

Cyclic AMP levels in isolated frog skin epithelium: effects of phosphodiesterase inhibitors, oxytocin and catecholamines.

Direct measurements of cyclic AMP were performed in the isolated epithelium of frog skin. Phosphodiesterase inhibitors (methylxanthines, papaverine) and activators of adenylyl cyclase (oxytocin, catecholamines) significantly increased the cyclic AMP content. Propranolol completely blocked the generation of cAMP induced by beta-adrenergic agonists but had little or no effect on that induced by oxytocin. Phentolamine enhanced the cAMP production by adrenalin and noradrenalin. At supramaximal concentrations, oxytocin and isoproterenol produced similar increments in cAMP, while exposure to both agents roughly doubled the increase in cAMP. The results suggest the presence of independent receptors for oxytocin and catecholamines in frog skin, with additive effects on cAMP generation.

Animals↗