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J. Biol. Chem., Vol. 263, Issue 13, 6155-6160, May, 1988
WK Sonnenburg and WL Smith
Prostaglandin E1 (PGE1) at 1 nM inhibits arginine-vasopressin (AVP)-
induced water reabsorption in the rabbit cortical collecting tubule (RCCT),
while 100 nM PGE1, by itself, stimulates water reabsorption (Grantham, J.
J., and Orloff, J. (1968) J. Clin. Invest. 47, 1154- 1161). To investigate
the basis for these two responses, we measured the effects of
prostaglandins on cAMP metabolism in purified RCCT cells. In freshly
isolated cells, PGE2, PGE1, and 16,16-dimethyl-PGE2 acting at high
concentrations (0.1-10 microM) stimulated cAMP accumulation; however, one
PGE2 analog, sulprostone (16-phenoxy- 17,18,19,20-tetranor-PGE2
methylsulfonilamide), failed to stimulate cAMP accumulation or to
antagonize PGE2-induced cAMP formation; PGD2, PGF2 alpha, and a PGI2
analog, carbacyclin (6-carbaprostaglandin I2), also failed to stimulate
cAMP synthesis. These results suggest that there is a PGE-specific
stimulatory receptor in RCCT cells which mediates activation of adenylate
cyclase. Occupancy of this receptor would be anticipated to cause water
reabsorption by the collecting tubule. At lower concentrations (0.1-100 nM)
PGE2, PGE1, 16,16-dimethyl- PGE2, and, in addition, sulprostone inhibited
AVP-induced cAMP accumulation by fresh RCCT cells in the presence of cAMP
phosphodiesterase inhibitors. Pertussis toxin pretreatment of RCCT cells
blocked the ability of both PGE2 and sulprostone to inhibit AVP- induced
cAMP accumulation. In membranes prepared from RCCT cells, sulprostone
prevented stimulation of adenylate cyclase by AVP. These results suggest
that E-series prostaglandins (including sulprostone) can act through an
inhibitory PGE receptor(s) coupled to the inhibitory guanine nucleotide
regulatory protein, Gi, to block AVP-induced cAMP synthesis by RCCT cells.
Occupancy of this receptor would be expected to cause inhibition of
AVP-induced water reabsorption in the intact tubule. Curiously, after RCCT
cells were cultured for 5-7 days, PGE2 no longer inhibited AVP-induced cAMP
accumulation, but PGE2 by itself could still stimulate cAMP accumulation.
In contrast to PGE2, epinephrine acting via an alpha 2-adrenergic,
Gi-linked mechanism did block AVP-induced cAMP formation by cultured RCCT
cells. This implies that some component of the inhibitory PGE response
other than Gi is lost when RCCT cells are cultured.
Regulation of cyclic AMP metabolism in rabbit cortical collecting tubule cells by prostaglandins
Department of Biochemistry, Michigan State University, East Lansing 48824.
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