ABSTRACT
Vasoactive intestinal peptide (VIP) receptors and beta-adrenergic receptors were investigated in rat Harderian gland membranes using 125I-VIP and 125I-cyanopindolol (125I-CYP), respectively, as ligands. The receptor bindings were rapid, reversible, saturable, specific, and dependent on time, temperature, and membrane concentration. The stoichiometric data suggested the presence of two classes of VIP receptors with Kd values of 0.36 and 65.37 nM and binding capacities of 323 and 39,537 fmol VIP/mg protein, respectively. The interaction showed a high degree of specificity, as suggested by competitive displacement experiments with several peptides structurally or not structurally related to VIP as follows: VIP > helodermin > rGRF > PHI > > secretin. Glucagon, somatostatin, insulin, and pancreastatin were ineffective at concentrations up to 1 microM. However, the stoichiometric data suggest the presence of one class of binding sites for 125I-CYP. The Kd for the single site was 290 pM with a binding capacity of 32 pmol/L. The pharmacological characterization of 125I-CYP binding to membranes showed that only isoproterenol, a beta-adrenergic agonist, and norepinephrine, an alpha beta-adrenergic agonist, was as effective as propranolol in inhibiting 125I-CYP binding to Harderian gland membranes. However, alpha 1- and alpha 2-adrenergic agonists and blockers such as methoxamine, prazosin, clonidine, and yohimbine were shown to be ineffective. These results demonstrate the presence of specific VIP and beta-adrenergic receptors in the Harderian gland and suggest a role for VIP and beta-adrenergic agonists in the physiology of this gland.
Subject(s)
Harderian Gland/metabolism , Receptors, Adrenergic, beta/metabolism , Receptors, Vasoactive Intestinal Peptide/metabolism , Animals , Binding Sites , Binding, Competitive , Male , Pindolol/agonists , Pindolol/analogs & derivatives , Pindolol/antagonists & inhibitors , Rats , Rats, Wistar , Vasoactive Intestinal Peptide/agonists , Vasoactive Intestinal Peptide/antagonists & inhibitorsABSTRACT
The disparate observations on the effect of halothane anesthesia on the serum electrolyte levels in humans prompted us to carry out this work. In this study the levels of sodium, potassium, magnesium, calcium, chloride and inorganic phosphorus were determined in serum of 25 male and 15 female patients, with an age range of 15 to 40 years, who had various pathologies requiring surgery and who were given halothane anesthesia. Significant difference were detected in the concentrations of sodium, potassium, calcium and inorganic phosphorus between presurgical and post-anesthesia induction samples. The truly striking finding in the present study was the significant increase in serum inorganic phosphorus in the intra-operative period. It is suspected that this increase is due to a defect in phosphorylating mechanisms which leads to a rapid hydrolysis of stored and preformed ATP.
Subject(s)
Anesthetics, Inhalation/pharmacology , Electrolytes/blood , Halothane/pharmacology , Preoperative Care/methods , Adolescent , Adult , Evaluation Studies as Topic , Female , Humans , MaleABSTRACT
Vasoactive intestinal peptide (VIP) receptors were investigated in rat Harderian gland membranes using [125I]VIP as ligand. The receptor binding was rapid, reversible, saturable, specific, and dependent on time, temperature, and membrane concentration. At 30 degrees C, the stoichiometric data suggested the presence of two classes of VIP receptors with Kd values of 0.36 +/- 0.06 and 65.37 +/- 8.08 nM and binding capacities of 323 +/- 54 and 39,537 +/- 3100 fmol VIP/mg protein, respectively. The interaction showed a high degree of specificity, as suggested by competitive displacement experiments with several peptides structurally or not structurally related to VIP. The binding of [125I]VIP to membranes was sensitive to guanine nucleotides in a dose-dependent manner. The molecular characterization of VIP receptors was realized by chemical cross-linking; sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the solubilized membrane proteins revealed the presence of two specific [125I]VIP-protein complexes of M(r) 57 and 35 kDa as estimated in denaturing conditions. VIP stimulated adenylate cyclase activity in rat Harderian gland membranes in a dose-dependent manner. Finally, VIP stimulated in vivo the type II thyroxine 5'-deiodinase activity. These results demonstrate the presence of specific and functional VIP receptors in Harderian gland and suggest a role for VIP in the physiology of this gland.
