Purinergic regulation of basal and arginine vasopressin-stimulated hydraulic conductivity in rabbit cortical collecting tubule

1985 ◽  
Vol 88 (3) ◽  
pp. 277-281 ◽  
Author(s):  
Mark A. Dillingham ◽  
Robert J. Anderson
Keyword(s):  
Hydraulic Conductivity ◽  
Arginine Vasopressin ◽  
Collecting Tubule ◽  
Rabbit Cortical Collecting Tubule ◽  
Cortical Collecting Tubule

Effects of calcium on ADH action in the cortical collecting tubule perfused in vitro

1982 ◽  
Vol 243 (5) ◽  
pp. F481-F486
Author(s):  
S. Goldfarb
Keyword(s):  
Hydraulic Conductivity ◽  
Water Flow ◽  
Collecting Tubule ◽  
Bath Water ◽  
Mammalian System ◽  
Rabbit Cortical Collecting Tubule ◽  
Cortical Collecting Tubule

To test the effects of calcium on ADH action in an in vitro mammalian system, the rabbit cortical collecting tubule was studied. After 25 microunits/ml ADH (n=8) in the presence of 1.25 mM calcium bath, water flow (Jv) rose to 1.56 +/- 0.34 nl.mm-1. min-1 and hydraulic conductivity (Lp, cm.s-1.atm-1 X 10(7)) rose to 123 +/- 22. After 25 microunits/ml ADH in the presence of 3.75 mM calcium bath (n=7), Jv rose to 2.96 +/- 0.6 nl.mm-1.min-1 (P less than 0.05 vs. control) and Lp rose to 286 +/- 62 cm.s-1.atm-1 X 10(7) (P less than 0.02 vs. 1.25 mM bath calcium control). Tubules (n=6) perfused with 3.75 mM Ca and bathed in 3.75 mM Ca also showed an Lp of 279 +/- 82 cm.s-1.atm-1 X 10 (7) following 25 microunits/ml ADH. Tubules similarly studied in 1.25 (n=6) or 3.75 mM Ca (n=6) bath but treated with 10(-4) M 8-[p-chlorophenylthio]cAMP demonstrated Lp of 222 +/- 26 and 235 +/- 37 cm.s-1.atm-1 X 10(7), respectively. These data suggest that increased bath Ca enhances ADH- but not cAMP-stimulated water flow in the rabbit cortical collecting tubule. High perfusate Ca2+ does not alter the stimulatory effect of elevated peritubular Ca2+.

Immunodissection and culture of rabbit cortical collecting tubule cells

1986 ◽  
Vol 251 (2) ◽  
pp. F348-F357
Author(s):  
W. S. Spielman ◽  
W. K. Sonnenburg ◽  
M. L. Allen ◽  
L. J. Arend ◽  
K. Gerozissis ◽  
...  
Keyword(s):  
Arginine Vasopressin ◽  
Renal Cortex ◽  
Morphological Properties ◽  
Prostaglandin D2 ◽  
Cortical Cells ◽  
Collecting Tubule ◽  
Tubule Cells ◽  
Principal And Intercalated Cells ◽  
Rabbit Cortical Collecting Tubule ◽  
Cortical Collecting Tubule

A mouse monoclonal antibody designated IgG3(rct-30) has been prepared that reacts specifically with an antigen on the surface of all cells comprising the cortical and medullary rabbit renal collecting tubule including the arcades. Plastic culture dishes coated with IgG3(rct-30) were used to isolate collecting tubule cells from collagenase dispersions of rabbit renal cortical cells by immunoadsorption. Typically, 10(6) rabbit cortical collecting tubule (RCCT) cells were obtained from 5 g of renal cortex (2 kidneys). Initial purity was greater than 96% based on immunocytofluorescent staining with three different anti-collecting tubule antibodies. Between 20 and 30% of the RCCT cells were reactive with peanut lectin suggesting that RCCT cells are a mixture of principal and intercalated cells. Approximately 10(7) RCCT cells were obtained after 4 to 5 days in primary culture. Moreover, RCCT cells continued to proliferate after passaging with a doubling time of approximately 32 h. RCCT cells passaged once and then cultured 4-5 days were found 1) to synthesize cAMP in response to arginine vasopressin (AVP), prostaglandin E2 (PGE2), isoproterenol, and parathyroid hormone, but not calcitonin, prostaglandin D2, or prostaglandin I, and 2) to release PGE2 in response to bradykinin but not arginine vasopressin or isoproterenol. Our results indicate that cultured RCCT cells retain many of the hormonal, histochemical, and morphological properties expected for a mixture of principal and intercalated rabbit cortical collecting tubule epithelia. RCCT cells should prove useful both for studying hormonal interactions in the cortical collecting tubule and as a starting population for isolating intercalated collecting tubule epithelia.

Amiloride-sensitive sodium channels in rabbit cortical collecting tubule primary cultures

1991 ◽  
Vol 261 (6) ◽  
pp. F933-F944 ◽  
Author(s):  
B. N. Ling ◽  
C. F. Hinton ◽  
D. C. Eaton
Keyword(s):  
High Selectivity ◽  
Primary Cultures ◽  
Principal Cell ◽  
Na Channel ◽  
Open Probability ◽  
Inward Current ◽  
Collecting Tubule ◽  
Apical Membranes ◽  
Rabbit Cortical Collecting Tubule ◽  
Cortical Collecting Tubule

Patch-clamp methodology was applied to principal cell apical membranes of rabbit cortical collecting tubule (CCT) primary cultures grown on collagen supports in the presence of aldosterone (1.5 microM). The most frequently observed channel had a unit conductance of 3-5 pS, nonlinear current-voltage (I-V) relationship, Na permeability (PNa)-to-K permeability (PK) ratio greater than 19:1, and inward current at all applied potentials (Vapp) less than +80 mV (n = 41). Less frequently, an 8- to 10-pS channel with a linear I-V curve, PNa/PK less than 5:1, and inward current at Vapp less than +40 mV was also observed (n = 7). Luminal amiloride (0.75 microM) decreased the open probability (Po) for both of these channels. Mean open time for the high-selectivity Na+ channel was 2.1 +/- 0.5 s and for the low-selectivity Na+ channel was 50 +/- 12 ms. In primary cultures grown without aldosterone the high-selectivity Na+ channel was rarely observed (1 of 32 patches). Lastly, a 26- to 35-pS channel, nonselective for Na+ over K+, was not activated by cytoplasmic Ca2+ or voltage nor inhibited by amiloride (n = 17). We conclude that under specific growth conditions, namely permeable transporting supports and chronic mineralocorticoid hormone exposure, principal cell apical membranes of rabbit CCT primary cultures contain 1) both high-selectivity and low-selectivity, amiloride-inhibitable Na+ channels and 2) amiloride-insensitive, nonselective cation channels.

Effects of AVP and dDAVP on PGE2 synthesis in superfused cortical collecting tubules

1989 ◽  
Vol 256 (6) ◽  
pp. F1044-F1050 ◽  
Author(s):  
F. Jaisser ◽  
L. Bugeon ◽  
M. Blot-Chabaud ◽  
J. P. Bonvalet ◽  
N. Farman
Keyword(s):  
Arginine Vasopressin ◽  
Dose Range ◽  
Maximal Effect ◽  
Pge2 Synthesis ◽  
Collecting Tubule ◽  
V2 Receptor ◽  
Collecting Tubules ◽  
Dose Dependent ◽  
Stimulation Of ◽  
Cortical Collecting Tubule

Whereas interactions between antidiuretic hormone (ADH) and prostaglandins (PGs) have been reported in the cortical collecting tubule (CCD), the precise effects of arginine vasopressin (AVP) and its analogue, 1-desamino-8-D-arginine vasopressin (dDAVP) on PGE2 synthesis remain controversial. We examined the dynamic response of PGE2 synthesis to these two analogues in isolated rabbit CCD. Microdissected CCD were superfused, and basal and hormone-induced PGE2 synthesis were determined by enzyme immunoassay. Addition of arachidonic acid (AA) steeply increased basal PGE2 synthesis, in the 0-1 microM-dose range. The presence of AA was necessary to obtain a stimulatory effect of AVP on PGE2 synthesis. AVP induced an immediate, transitory, and dose-dependent stimulation of PGE2 synthesis. A maximal effect was obtained at 10(-8) M; PGE2 synthesis was increased by approximately 150-200% over the basal synthesis. With dDAVP, a very weak response was obtained only at 10(-7) M. From these results, we conclude that PGE2 synthesis in CCD is stimulated by ADH. This effect of ADH does not depend on the V2-receptor pathway and suggests the presence of V1-receptors in CCD.

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