1,721,400 research outputs found
Protective effect of hydrocortisone on vasopressin response in frog skin
No full textThe effect of microtubular-poisons, such as colchicine and vincristine, on frog skin permeability has been investigated. Three-hour treatment with the drugs has no effect on nonelectrolyte basal transepithelial permeability, but completely suppresses the effect of ADH. Colchicine and vincristine, in addition, affect both basal sodium transport and the rise in short circuit current induced by vasopressin. The inhibition produced by microtubular-poisons disappears, however, when hydrocortisone, a glucocorticoid known to preserve junctional communications is used. Together with the results previously obtained with isolated epithelial cells (Svelto et al. 1979), these findings provide further support for our hypothesis that the microtubular-microfilament-system, is involved in cell-to-cell exchange
Calcium role in base-line and ADH-stimulated sodium transport in frog skin
No full textTreatment of ventral frog skin with serosal A23187 calcium ionophore caused an initially transient increase in transepithelial sodium transport. After 60 min of treatment with A23187, a steady-state transport value was reached which was significantly lower than the initial one. Furthermore, it was found that ionophore treatment greatly inhibited the natriferic response to ADH and to 8br-cAMP. A further analysis on the possible ionophore action mechanism was carried out through pretreatment of the skin with indomethacin, very powerful prostaglandin synthesis inhibitor. In the experimental conditions reported, A23187 seems no longer capable of inducing a transient increase in sodium transport, although it does inhibit the natriferic response to ADH
Colchicine inhibition of ADH effect on frog skin permeability.
No full textADH and AMPc enhance both thiourea unidirectional fluxes in frog skin. This effect is completely abolished by colchicine pretreatment. The ADH increase of thiourea discharge with or without colchicine led us to suppose that colchicine does not directly affect ADH action on outer membrane permeability, but exerts its effects on a site which is limiting for the ADH action on transepithelial permeability
Evidence for the role of calcium in the hydrosmotic response to antidiuretic hormone in frog skin
No full textTreatment with the calcium ionophore A23187 on either the serosal or mucosal sides of frog skin, strongly inhibits the hydrosmotic response to vasopressin. On the contrary, the hydrosmotic response to 8-br-cAMP is not affected by treatment with the A23187. Trifluoperazine, a drug which inhibits the Ca2+-calmodulin complex, selectively inhibits vasopressin-induced water transport. Collectively, our results suggest that an increase in the intracellular concentration of Ca2+, obtained by treatment with the ionophore A23187, interferes with a pre-cAMP step of the hydrosmotic response to the antidiuretic hormone. Calcium ions could regulate adenyl-cyclase activity and consequently intracellular levels of cAMP. This effect may probably involve calmodulin
Noradrenaline induced secretion of nonelectrolytes through frog skin
No full textAddition of noradrenaline (4 x 10(-5) M) to the inner bathing fluid in the skin of the frog Rana esculenta results in increased unidirectional fluxes of urea, thiourea, N-methyl-thiourea, N-N'-dimethylthiourea and mannitol. Fluxes towards the external medium (phi o) undergo a much greater increase than those moving the opposite direction (phi i). The effect of noradrenaline on phi o is higher for urea and thiourea than mannitol, while its effect on phi o thiourea derivatives is related to lipid solubility. This phenomenon does not occur for phi i of the same molecules. FCCP (10(-6) M) pretreatment strongly inhibits the noradrenaline effect on phi o. In skin pretreated with colchicine (2 x 10(-5) M) both urea fluxes are increased to the same extent by noradrenaline. Noradrenaline is concluded to exert two separate effects: (1) a change in permeability in both directions; (2) a secretion of nonelectrolytes towards the external fluid. Such secretion is most probably associated with the hormone-induced secretion of fluid and electrolytes, perhaps mediated by an exocytotic mechanism
Possibile functional implications of aquaporin water channels in reproductive physiology and medically assisted procreation
No full textSpermatogenesis, the maturation of spermatozoa and their concentration and storage in the seminiferous vessels are associated with considerable fluid secretion or absorption in the male reproductive tract. These fluid movements are in total agreement with the presence of multiple aquaporin (AQP) water channel proteins in germ cells and other tissues within the male reproductive tract. A series of functions of prime importance have already been hypothesized for aquaporins in the physiology of male reproduction. Aquaporins could be involved in the early stages of spermatogenesis, in the secretion of tubular liquid and in the concentration and storage of spermatozoa in the epididymis. In the male reproductive tract, alterations in the expression and functionality and/or regulation of aquaporins have already been demonstrated to be at the basis of forms of male infertility. Indeed, rats with reduced reabsorption of seminiferous fluid in the efferent ducts have been shown to be sub-fertile or infertile. Functions have also been suggested in the fertilization process, where aquaporins may play a role in maintaining osmotic homeostasis in gametes during.-e fertilization. Aquaporins have also been suggested to mediate water movement into antral follicles and to be the pathway for transtrophectodermal water movement during cavitation. Aquaporins are the subject of considerable technological interest for cryopreservation used in medically assisted procreation, as they could be the molecular pathway by which water and/or solutes move across the plasma membrane during the process of freezing/thawing gametes and embryos. Indeed, artificial expression of AQP3 has been showed to improve the survival of mouse oocytes after cryopreservation
Recruitment of F-actin and the actin-binding protein moesin in the membrane protrusion during cell swelling
No full textPolyclonal antibodies in study of ADH-induced water channels in frog urinary bladder.
No full textIt is now generally accepted that changes in water permeability in anti-diuretic hormone (ADH)-responsive target epithelial cells result from the insertion in the plasma apical membrane of new components that contain channels for water. The specificity of these channels suggests that they are formed by intrinsic proteins having access to both facies and spanning the whole membrane. We have previously shown that Triton X-100 apical extracts from ADH-stimulated frog urinary bladder contain some proteins inserted under hormonal stimulation. In the present study we have developed polyclonal antibodies using Triton X-100 extract as an immunogen. After considering the inhibitory effect exerted by the whole immune serum on the osmotic water flow, we used different adsorption steps to select, from the immune serum, antibodies to apical membrane proteins inserted in response to the hormone. Immunoblot analysis of these selected antibodies shows that they recognize seven to eight proteins, of which 55-, 35-, 26-, and 17-kDa proteins are always present. Antibodies to these four proteins, affinity purified on nitrocellulose sheets, inhibited ADH-induced osmotic water flow. Altogether these results strongly suggest that proteins of 55, 35, 26, and 17 kDa (or at least one of them) are likely to be involved in the mechanism of water transport
A comparative structural analysis of the central (fifth) pore of aquaporins: electrostatic features suggest different conductance properties for different aquaporin subfamilies
No full textAquaporins (AQPs) are homotetrameric channel proteins allowing the diffusion of water/small solutes across membranes. AQP structures are very similar, with each monomer defining a single pore selective to water/solutes and contributing to form a fifth central pore, whose meaning remains elusive. Nevertheless, AQPs show distinct transport selectivity to water, orthodox AQPs, or glycerol/solutes, aquaglyceroporins. The variety of available AQP 3D-structures is a valuable resource for studying the structure-function relationships within this protein family. A recent comparative analysis allowed specific electrostatic profiles to be associated with the main AQP selectivity to water and glycerol. We exploited this approach to gain some insights into the role of the AQP central pore. Interestingly: the electrostatics of AQP central channels correlates with their main transport function; AQP1 and AQP4 fifth pore has strikingly comparable electrostatics, supporting previous works reporting its involvement in the transport of CO2 across membranes; the central pore of the spinach PIP2;1 shares the same electrostatic profile of the monomeric pore of orthodox AQPs, suggesting that the fifth pore could allegedly represent an alternative/additional path for the transport of water across (at least some) plant AQPs. The hypothesis is being verified experimentally
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