atrial-natriuretic-factor and calmidazolium

Natriuretic peptides (NPs) may work as neuromodulators through their associated receptors [NP receptors (NPRs)]. By immunocytochemistry, we showed that NPR-A and NPR-B were expressed abundantly on both ON-type and OFF-type bipolar cells (BCs) in rat retina, including the dendrites, somata, and axon terminals. Whole-cell recordings made from isolated ON-type BCs further showed that brain natriuretic peptide (BNP) suppressed GABAA receptor-, but not GABAC receptor-, mediated currents of the BCs, which was blocked by the NPR-A antagonist anantin. The NPR-C agonist c-ANF [des(Gln18, Ser19, Gln20, Leu21, Gly22)ANF(4-23)-NH2] did not suppress GABAA currents. The BNP effect on GABAA currents was abolished with preincubation with the pGC-A/B antagonist HS-142-1 but mimicked by application of 8-bromoguanosine-3',5'-cyclomonophosphate. These results suggest that elevated levels of intracellular cGMP caused by activation of NPR-A may mediate the BNP effect. Internal infusion of the cGMP-dependent protein kinase G (PKG) inhibitor KT5823 essentially blocked the BNP-induced reduction of GABAA currents. Moreover, calcium imaging showed that BNP caused a significant elevation of intracellular calcium that could be caused by increased calcium release from intracellular stores by PKG. The BNP effect was blocked by the ryanodine receptor modulators caffeine, ryanodine, and ruthenium red but not by the IP3 receptor antagonists heparin and xestospongin-C. Furthermore, the BNP effect was abolished after application of the blocker of endoplasmic reticulum Ca2+-ATPase thapsigargin and greatly reduced by the calmodulin inhibitors W-7 and calmidazolium. We therefore conclude that the increased calcium release from ryanodine-sensitive calcium stores by BNP may be responsible for the BNP-caused GABAA response suppression in ON-type BCs through stimulating calmodulin.

Topics: Animals; Atrial Natriuretic Factor; Caffeine; Calcium; Calcium Channels; Calcium Signaling; Calcium-Transporting ATPases; Calmodulin; Carbazoles; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; GABA-A Receptor Antagonists; gamma-Aminobutyric Acid; Guanylate Cyclase; Heparin; Imidazoles; Indoles; Inositol 1,4,5-Trisphosphate Receptors; Macrocyclic Compounds; Male; Membrane Potentials; Natriuretic Peptide, Brain; Oxazoles; Patch-Clamp Techniques; Peptide Fragments; Peptides, Cyclic; Polysaccharides; Protein Kinase Inhibitors; Rats; Rats, Sprague-Dawley; Receptors, Atrial Natriuretic Factor; Receptors, Cytoplasmic and Nuclear; Receptors, GABA; Receptors, GABA-A; Retinal Bipolar Cells; Ruthenium Red; Ryanodine; Ryanodine Receptor Calcium Release Channel; Thapsigargin

Effects of 8-bromo-cGMP (8-Br-cGMP) and synthetic rat atriopeptin III (APIII) on sodium absorption by isolated chicken villus enterocytes and intact chicken ileal mucosa were determined. In isolated cells, both agents significantly decreased initial rates of influx of 22Na and caused a persistent decrease in intracellular pH (pHi); effects that are not additive to those caused by amiloride (10(-3) M). The ED50 for APIII was 0.3 nM. In intact mucosa, both 8-Br-cGMP (10(-4) M) and 5-(N-methyl-N-isobutyl)amiloride (MIBA) (10(-5) M) reduced JNams and JNa.net, their effects were not additive. APIII (10(-7) M) significantly increased cellular cGMP but not cAMP. Both 8-Br-cGMP (10(-4) M) and APIII (10(-7) M) stimulated a persistent increase in cytosolic calcium (Cai), which could be prevented by pretreating the cells with the cytosolic calcium buffering agent MAPTAM or with H-8, an inhibitor of cyclic nucleotide-dependent protein kinases. Furthermore, pretreatment of cells with H-8 or the calmodulin inhibitor, calmidazolium (CM), prevented the effects of 8-Br-cGMP and APIII on pHi. However, the pHi response to subsequent addition of the calcium-ionophore ionomycin was blocked only by CM and not by H-8. These data suggest that APIII and 8-Br-cGMP inhibit amiloride-sensitive Na/H exchange by increasing Cai, an event requiring activation of cGMP-dependent protein kinase.

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Amiloride; Animals; Atrial Natriuretic Factor; Biological Transport; Calcium; Carrier Proteins; Chickens; Cyclic GMP; Hydrogen-Ion Concentration; Ileum; Imidazoles; In Vitro Techniques; Intestinal Absorption; Intestinal Mucosa; Isoquinolines; Sodium; Sodium-Hydrogen Exchangers

Atrial natriuretic factor (ANF) was studied for its effect on renin release by rat renal cortical slices. ANF (rat) alone (10(-9)-10(-6) M) had no effect on basal renin release, but significantly (P less than 0.001) potentiated angiotensin II (AII) inhibition of renin secretion in doses as low as 10(-9) M. ANF also potentiated the inhibitory effect of AII on 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride (which alters intracellular calcium) and calmidazolium (a calmodulin blocker) effects on renin release. ANF did not inhibit the action of isoproterenol, which probably acts through cAMP, at doses below micromolar concentrations. Large, probably pharmacological amounts (greater than 10(-6) M) produce some inhibition. Since AII action on renin is associated with increases in intracellular calcium, our studies suggest that ANF acts by altering the intracellular calcium-calmodulin-mediated steps of AII action on this tissue and not via cAMP.

Topics: Angiotensin II; Animals; Atrial Natriuretic Factor; Depression, Chemical; Drug Synergism; Gallic Acid; Imidazoles; Isoproterenol; Kidney Cortex; Male; Rats; Rats, Inbred Strains; Renin; Secretory Rate