Effects of atrial natriuretic factor and vasopressin on cyclic nucleotides in cultured kidney cells.

Abstract

The cellular mechanism of the action of atrial natriuretic factor (ANF) is thought to involve activation of guanylate cyclase. Increasing evidence shows a direct tubular effect of ANF. Part of the ANF-induced diuresis has been suggested to be due to inhibition of the action of arginine vasopressin (AVP) in the cortical collecting tubule. In this study we investigated the effect of ANF on cyclic nucleotide production in primary cultures of cortical collecting tubule cells immunodissected with a monoclonal antibody. ANF caused a dose-dependent stimulation in cyclic guanosine 3',5'-monophosphate (cGMP) production; the half-maximal stimulation was observed at approximately 1 nM of ANF. ANF (0.01-100 nM) had no effect on cyclic adenosine 3',5'-monophosphate (cAMP) accumulation in cortical collecting tubule cultures. AVP caused a dose-dependent increase in cAMP production, and this effect was not altered by the simultaneous addition of ANF (100 nM). Similarly, ANF-induced cGMP stimulation was not influenced by AVP (10 nM). We conclude that 1) ANF has a direct stimulatory action on cGMP production by cultured cortical collecting tubule cells and 2) any interaction between ANF and AVP is likely to occur at steps distal to cyclic nucleotide formation.

Culture of human renal cortex epithelial cells

“…Although sophisticated methodologies to prepare segmentspecific cultured ceils have emerged, technical limitations restrict their applicability. For example, the "immunodissection" technique (6,(13)(14)(15) used to isolate cells that share a common cell type-specific ectoantigen is an important methodologic advancement with considerable promise, but it is technically rigorous and demands fastidious immunologic and cell type characterization, beyond the grasp of many laboratories, to be successful. Microdissection of selected nephron segments for culture (3,7,19,20), likewise, is a very promising methodology and has been applied to human kidney (21).…”

Post-Doctoral Research Award

“…In this portion of the study, we provided evidence that CCD ceUs absorb calcium via active transport mechanisms and that a Naf/H÷ exchange mechanism could be involved. CCD cells isolated and cultured for this study retained characteristics of CCD cells in vivo including absorption of sodium and chloride, secretion of potassium and protons (1,2,3) and responsiveness to the hormones aldosterone and vasopressin (3,4). These cells, when cultured on a permeable filter, formed a tight monolayer as demonstrated by the high transepithelial voltages (60 -70 mV) and resistances (greater than 1000 ohm /cm 2 ) obtained.…”

“…4 A technique for concentrating released NO in the cell medium is to collect samples eluted from a column of cells grown on beads. Endothelial cells seem to produce very small quantities of NO when stimulated (12).…”

Effect of Atrial Natriuretic Factor on Vasopressin-Stimulated Cyclic AMP Accumulation in Papillary Collecting Tubule

“…cAMP generation [8], In cultured rabbit cor tical collecting tubule cells, ANF had no effect on basal or AVP-stimulated cAMP generation [9], In rabbit outer and inner me dullary tubule suspensions, and in rat microdissected inner medullay collecting ducts, ANF had no effect on AVP-stimulated cAMP generation [10], In homogenates of dog medullary collecting ducts, ANF inhibit ed basal adenylate cyclase activity, but had no effect on AVP-stimulated adenylate cy clase activity [11], Finally, in rat cultured papillary collecting tubule cells, ANF had no effect on basal cAMP accumulation [12], ANF did inhibit AVP and forskolin-stimulated cAMP accumulation; however, most studies required at least 10~7to 10-6 M ANF to see an effect.…”