pituitrin and inositol-1-4-bis(phosphate)

The hydrolysis of inositol phospholipids induced by vasopressin in hepatocytes during 60 min was quantified chemically. There was a large release of myo-inositol which was abolished by Li+, indicating that it was derived from inositol phosphates and not from phospholipase D action on PtdIns. There was also a large release of inositol phosphates which was increased approx. 2-fold by Li+ at 30 min, but then remained constant, suggesting that inositol phospholipid breakdown declined substantially beyond this time. In cells prelabelled with myo-[3H]inositol and treated with Li+, [3H]PtdIns(4,5)P2 decreased maximally (50%) at 15 s and then recovered to a level at 5 min that was maintained at 25% below control for 40 min. [3H]PtdIns4P and [3H]PtdIns showed slower decreases to approx. 30% below control at 15 min, but with no further changes. Labelled Ins(1,4,5)P3 and Ins(1,3,4)P3 showed 2-4-fold increases within 30 s and then declined to values that were maintained at a constant level above the control, except for [3H]Ins(1,3,4)P3, which showed a second increase. [3H]Ins(1,4)P2 showed a very large increase over 10 min, whereas [3H]Ins4P and [3H]Ins1P showed little change before 6 and 15 min respectively. The total [3H]inositol phosphates showed little further increase after 20 min. These data are consistent with a rapid, but not sustained, hydrolysis of PtdIns-(4,5)P2, but not of PtdIns, by phospholipase C, but do not exclude PtdIns4P as a substrate. Phosphatidate was rapidly increased by vasopressin, whereas diacylglycerol was increased after a 1-2 min lag. Both were maintained at levels 2-3-fold above control for 60 min. The vasopressin-induced increase in inositol phosphates plus myo-inositol (approx. 120 nmol/100 mg) was greater than the increase in diacylglycerol plus phosphatidate (approx. 60 nmol/100 mg) between 10 and 40 min. This indicates that there was substantial further metabolism of these lipids. Addition of 75 mM ethanol resulted in rapid production of phosphatidylethanol in response to vasopressin and a 35% reduction in phosphatidate, but no decrease in diacylglycerol. In summary, the results indicate that inositol phospholipid hydrolysis by phospholipase C can account for most of the diacylglycerol and phosphatidate that accumulate during 60 min of vasopressin action, but that these phospholipids are probably not the major source of the phosphatidate that is formed during the first 2 min by phospholipase D, or of the diacylglycerol and phosphati

Topics: Animals; Diglycerides; Ethanol; Hydrolysis; Inositol; Inositol 1,4,5-Trisphosphate; Inositol Phosphates; Kinetics; Lithium; Liver; Male; Phosphatidic Acids; Phosphatidylinositols; Rats; Rats, Sprague-Dawley; Vasopressins

We have previously shown that vasopressin exerts a marked mitogenic effect on adrenal glomerulosa cells. In the present study, we demonstrate that vasopressin (VP) stimulates the formation of inositol monophosphate (IP), inositol diphosphate (IP2) and inositol triphosphate (IP3) in primary cultures of glomerulosa as well as fasciculata cells 5- to 8-fold over the corresponding basal values. In both cell types, the relative stimulations of IP, IP2, and IP3 formation were similar. Angiotensin II (ATII) also induced glomerulosa cells to produce a dose-dependent (up to 10-fold) increase in IP, IP2, and IP3, but had only a small effect on fasciculata cells. The dose dependencies for ATII-induced IP, IP2, and IP3 formation and aldosterone production were nearly the same. We conclude that VP- and ATII-induced formation of inositol phosphates may represent an early step in the action of these peptides on adrenal cells. However, additional elements must be involved to account for the cell specificity of VP and ATII. In glomerulosa cells, VP stimulates mitotic activity and aldosterone secretion, while ATII is only steroidogenic. On fasciculata cells, VP induces a significant increase in the formation of inositol phosphates in spite of the absence of a known biological function in these cells.

Topics: Adrenal Cortex; Angiotensin II; Animals; Dose-Response Relationship, Drug; Inositol 1,4,5-Trisphosphate; Inositol Phosphates; Male; Membranes; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositol Phosphates; Phosphatidylinositols; Rats; Time Factors; Vasopressins