European Journal of PharmacoloD', 109 (1985) 249-256 Elsevier

249

INCREASE OF P L A S M A R E N I N ACTIVITY AFTER S U B C U T A N E O U S A P P L I C A T I O N OF C O M P O U N D 4 8 / 8 0 IN THE RAT HIROSHI IZUMI *, SIU-CHEONGHO *, ANDREW M. MICHELAKIS* and TSUYOSHI AOKI
Department of Physiology, Tohoku Unit,ersi(t' School of Dentisto', Seio'o-Machi 4-1, Sendai 980, Japan, and * Department of Pharmacology and Toxicology, Michigan State Unit,ersitv, East Lansing, Michigan 48824, U.S.A.

Received 13 June 1984. revised MS received 1 November 1984, accepted 27 November 1984

H. IZUMI, S.-C. HO, A.M. MICHELAKIS and T. AOKI, Increase of plasma renin activity after subcutaneous application of compound 4 8 / 8 0 in the rat, European J. Pharmacol. 109 (1985) 249-256. Subcutaneous (s.c.) administration of compound 48/80 (3.0 mg/kg) to conscious rats produced a time-dependent long-lasting increase of plasma renin activity (PRA). A dose-related increase of the hematocrit was also observed after injection of compound 48/80. The onset of the hematocrit increase preceded that of PRA increase. Pretreatment with a dose of more than 20 mg/kg of histamine HI-receptor antagonists such as tripelennamine or diphenhydramine prior to the injection of compound 48/80 (3.0 mg/kg s.c.) attenuated or abolished the effects of compound 48/80 on PRA, hematocrit and plasma extravasation. Pretreatment with cimetidine (histamine H2-receptor antagonist, 40 mg/kg i.p.) had no effect on these plasma variables. The increase of PRA caused by s.c. administration of compound 48/80 was not affected by the pretreatment with propranolol (fl-adrenoceptor antagonist, 10 mg/kg i.p.), which completely inhibited the isoproterenol (0.5 mg/kg s.c.)-induced PRA increase. Administration of compound 48/80 did not induce a significant PRA increase in the nephrectomized rats although the increase of hematocrit following s.c. administration of compound 48/80 persisted despite the absence of kidneys. S.c. administration of compound 48/80 (3.0 mg/kg) led to a significant decrease of histamine content at the site of injection and to a significant increase in plasma histamine concentration without affecting arterial blood pressure. The present data suggest that s.c. administration of compound 48/80 stimulates the release of histamine from cutaneous mast cells, which causes an increase in vascular permeability to plasma protein via the stimulation of histamine HI-receptors, then leads to hypovolemia. The resulting hypovolemia may directly stimulate the juxtraglomerular cells of the kidney to release renin. Compound 48/80 Hypovolemia Plasma renin activity Plasma extravasation Hematocrit Histamine HI-receptor

1. Introduction
The loss of blood plasma volume (induction of hypovolemia) by either administration of polyethylene glycol or water deprivation is known to stimulate the increase of angiotensin II in plasma and to induce drinking (Kutscher, 1968; Malvin et al., 1977; Vijande et al., 1978; Mann et al., 1980; Johnson et al., 1981; Yamaguchi et al., 1982). However, little information is available concerning the relationship between plasma extravasation and To whom all correspondenceshould be addressed at Tohoku University. 0014-2999/85/$03.30 1985 Elsevier Science Publishers B.V.

the renin-angiotensin system, although intraperitoneal (i.p.) or intravenous (i.v.) injection of kidney extract or renin into nephrectomized rats has been reported to cause not only thirst but also leakage of plasma protein from the vascular compartment (Paldino and Hyman, 1954; Asscher and Anson, 1963; Giese, 1963; Fitzsimons, 1969; Haefeli and Peters, 1971). It was also suggested that release of histamine from mast cells may be involved in neurogenic plasma extravasation, antidromic vasodilation, reactive hyperemia and food-related drinking (Kiernan, 1975; Arvier et al., 1977; Lembeck and Holzer, 1979; Erjavec et al., 1981; Kraly, 1983).

250 We have recently reported that subcutaneous (s.c.) administration of compound 48/80, a wellknown histamine liberator from various tissues (Paton, 1951; Feldberg and Talesnik, 1953; Johnson and Erd6s, 1973; Douglas, 1980; Gristwood et al., 1981), produced a marked increase of plasma renin activity (PRA) and drinking in rats, although histamine itself had no such effect on PRA when given s.c. to rats (Izumi et al., 1983b). The purpose of the present studies was to examine the mechanism mediating the increase of PRA in response to the s.c. administration of compound 4 8 / 8 0 in conscious rats. determined by the radioimmunoassay method of Haber et al. (1969). PRA was expressed as nanograms of angiotensin I generated at 37C minus nanograms of angiotensin I at 0C per ml of plasma per 3 h of incubation.

2.4. Determination of arterial blood pressure

Rats were anesthetized with sodium pentobarbital (50 m g / k g i.p.) supplemented with ether. After inserting a PE 10 tube into the femoral artery, one end of the catheter was passed through the abdominal muscle and led out s.c. to exit through the dorsal neck skin. Two to 3 days later, arterial blood pressure was recorded in the conscious rats from the femoral artery catheter with a Grass Polygraph (Model 70) and a Statham pressure transducer.

2. Materials and methods

2.1. Animal maintenance

Normal male rats weighing 200-220 g of a Wistar strain were housed, 3 per cage, in a temperature (20-22C) - and humidity (55%)-controlled room on a 12:12 h light-dark cycle. Commercial food and water were available ad libitum.

2.5. Determination of plasma extravasation

A 2% solution of Evans blue in 0.9% NaC1, 2.5 m l / k g , was injected into the tail vein according to the method described by Arvier et al. (1977) and 5 min afterwards compound 48/80 or histamine were administered s.c. After 30 min, skin at the site of injection (100 mg wet weight) was removed and cut into small pieces with scissors. Extravasated Evans blue was extracted for colorimetric determination as described by Harada et al. (1971).

2.2. Collection of blood

One h after the injection of drug, unless otherwise stated, the animals were killed by decapitation within 10 s of opening the cage to minimize stress, and trunk blood (3.0 ml) was collected into a chilled centrifuge tube, containing 0.3 ml of isotonic saline with 0.1 M EDTA, and centrifuged at 4C. Plasma was stored at - 20C until assayed for PRA and histamine concentration.

2.6. Determination of histamine concentration in plasma and tissues

The histamine assay with H P L C has been described in detail (Izumi et al., 1984). Briefly, after separation of histamine from other amines by the cellulose phosphate column method described by Endo (1981), histamine was measured by the HPLC-fluorescence analysis system. This system consists of a Waters Model 204 Liquid Chromatograph with a 6000A Solvent System, a U 6 K Universal Injector, Radial Pack A column (Waters Associates Inc., Milford, MA, U.S.A.), and a Jasco-FP-550 Spectrofluorometer (Japan Spectroscopic Co., LTD, Tokyo, Japan) equipped with a flow-cell unit (cell volume 15 ~1). The mobile phase was a mixture of 0.5 M phosphate buffer

2.3. PRA determination

Aliquots of 0.1 ml of test plasma were incubated for 3 h at 37C and 0C in maleate buffer (pH 6.5) containing 6.0 m M E D T A and 2.0 mM phenylmethanesulfonyl-fluoride, in the presence of 48 h nephrectomized rat plasma as renin substrate (Izumi et al., 1982, 1983a). The nephrectomized rat plasma contained in excess of 5077.6 ng angiotensin I equivalent renin substrate per ml compared to 571.9 ng for normal plasma. The angiotensin I generated at both 37C and 0C incubations was

251

(pH 4.7; 75 ml), triethylamine (0.38 ml), 0.2 M HCI (15 ml) and methanol (90 ml). The flow rate was set at 1.0 ml/min. The fluorescence intensity was monitored at Ex.360/Em.440 nm. 2.7. Surgical removal of the kidneys Nephrectomy was performed under light ether anesthesia through dorsal incisions. The rats were given water and food ad libitum. The experiments studying the effects of compound 48/80 on PRA were performed 2 h after nephrectomy. 2.8. Determination of hematocrit Hematocrit measurements were done in triplicate by the microcapillary method using trunk blood collected after decapitation. 2.9. Drug administration Compound 48/80 solutions were prepared daily in 0.9% NaC1 prior to use, avoiding light exposure. Histamine, isoproterenol, propranolol, tripelennamine and diphenhydramine were dissolved in 0.9% NaC1. Cimetidine was dissolved in a minimal

quantity of 0.5 N HC1 and neutralized with 0.5 N NaOH to attain a pH of 6.0 then diluted to an appropriate volume with 0.9% NaC1. Aliquots of solutions of compound 48/80, isoproterenol and physiological saline (control) were injected s.c. Propranolol, tripelennamine, diphenhydramine and cimetidine were injected i.p. 20, 30, 30 and 30 min, respectively, before the administration of compound 48/80 and isoproterenol. 2.10. Statistics The significance of differences between group means was tested using Student's t-test. The absence of an asterisk indicates that the apparent difference was not significant at the P = 0.05 level. 2.11. Materials Compound 48/80 (N-methylhomoanisylamineformaldehyde condensation product), DL-isoproterenol hydrochloride, DL-propranolol hydrochloride, diphenhydramine hydrochloride and cimetidine were obtained from Sigma Chemical Co. St. Louis, MO, U.S.A. Tripelennamine hydrochloride was purchased from Ciba-Geigy Corp., Basel, Switzerland.

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252

3. Results

3.1. Time course of PRA and hematocrit response to compound 4 8 / 8 0 in normal rats
After s.c. administration of compound 4 8 / 8 0 (3.0 m g / k g ) to the conscious rats the blood was collected at various times, and PRA and hematocrit were measured at each time (fig. 1). A marked increase of PRA was observed within l0 min of compound 48//80 injection. A significant increase of hematocrit or PRA could be detected 5 or 10 min after s.c. administration of compound 4 8 / 8 0 to the rats. The changes of these plasma variables

were time-dependent and lasted up to 2 h. They then began to decline and returned toward normal.

3.2. Effect of compound 4 8 / 8 0 on tripelennamine-, diphenylhydramine-, cimetidine- and propranololpretreated rats

Fig. 2 shows the responses of PRA and hematocrit to s.c. administration of compound 48/80, histamine and isoproterenol, and the effects of tripelennamine, diphenylhydramine (both are histamine HI-receptor antagonists), cimetidine (histamine H2-receptor antagonist) and proprano-

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Fig. 2. Effects of tripelennamine, diphenhydramine, cimetidine and propranolol on compound 48/80-induced increase of plasma renin activity (PRA) and hematocrit. Aliquots of compound 48/80 (0.5, 1.0 and 3.0 mg/kg), isoproterenol (0.5 mg/kg), histamine (4 mg/kg) and physiological saline (control) were injected s.c. Tripelennamine (10, 20, 40 mg/kg), diphenhydramine (10, 20, 40 mg/kg), cimetidine (40 mg/kg) and propranolol (10 mg/kg) were administered i.p. 30, 30, 30 and 20 min, respectively, before the injection of compound 48/80 and isoproterenol. Open and dotted columns represent the PRA and the hematocrit, respectively. PRA was determined 1 h after the injection of compound 48/80, isoproterenol and histamine as described in Materials and methods and is expressed as ng angiotensin I produced by one ml of plasma during 3 h incubations. Results are expressed as means _+S.E.M. The figure in the bars indicates the number of rats. The numbers in parentheses indicate the concentration of drug used. Asterisks indicate a statistical difference from the control. * P < 0.05; ** P < 0.001. The letters in parentheses represent the statistical differences between rats treated with compound 48/80 (3 mg/kg) and those given compound 48/80 (3 mg/kg) and tripelennamine, diphenhydramine or cimetidine. (a) P < 0.05; (b) P < 0.01; (c) P < 0.001.

253

TABLE 1 Effects of sham-nephrectomy or nephrectomy on plasma renin activity (PRA) and hematocrit (Hct) after s.c. administration of compound 48/80 (C48/80, 3.0 mg/kg). PRA is expressed as ng angiotensin I produced by 1 ml plasma during 3 h incubations. Results are expressed as means + S.E.M. Number of rats is given in parentheses. Asterisks indicate a statistical differences from control. PRA Hct (%)

Control (6) 8.77 + 1.32 42.85 + 1.20 Sham (6) 6.07+ 0.88 42.05+0.73 Sham + C48/80 (9) 185.31 + 14.81 ** 55.30_+ 1.80 ** Nephrectomy (2 h) (9) 2.91 _+ 2.53 * 41.00+0.81 Nephrectomy + C48/80 (9) 3.64 + 2.78 * 53.45 _+1.35 ** * P < 0 . 0 1 ; ** P<0.001.

variables were not affected by s.c. injection of histamine (4.0 mg/kg). The above responses with compound 48/80 (3.0 mg/kg) were attenuated by the pretreatment with relatively larger doses of tripelennamine or diphenylhydramine but not with low dose of tripelennamine (10 mg/kg i.p.). Cimetidine (40 mg/kg i.p.) had no effect on either of the responses. The response of PRA to the s.c. administration of compound 48/80 (3.0 mg/kg) was not affected by the pretreatment with propranolol (10 mg/kg i.p.), which completely inhibited the isoproterenol (0.5 mg/kg s.c.)-induced PRA increase.
3.3. Effect of compound 4 8 / 8 0 on PRA and hematocrit in nephrectomized rats

lol (/3-adrenoceptive antagonist) on the responses of the above plasma variables to compound 48/80. The dose-dependent increases of PRA and hematocrit were observed 1 h after s.c. administration of compound 48/80. The values of these plasma
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Two h after nephrectomy, PRA was significantly reduced compared to that in control or sham-operated rats (table 1). Administration of compound 48/80 did not induce a significant PRA increase in nephrectomized rats although the compound 48/80 (3.0 mg/kg)-induced increase of hematocrit was observed despite the absence of the kidneys. No significant difference of hematocrit was observed between control or sham-operated rats and nephrectomized rats.
3.4. Effects of compound 4 8 / 8 0 on plasma extravasation

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Fig. 3 shows the effects of s.c. administration of compound 48/80 and histamine on plasma extravasation, and the effects of tripelennamine and cimetidine on the responses of plasma extravasation by compound 48/80. Plasma extravasation caused by 3.0 mg/kg compound 48/80 was much higher than that caused by 0.5 mg/kg compound 48/80. Plasma extravasation caused by compound 48/80 (3.0 mg/kg) was reduced by the pretreatment with tripelennamine but not by cimetidine. There was a small but significant plasma extravasation after s.c. administration of histamine (4.0 mg/kg).
3.5. Effects of compound 4 8 / 8 0 on histamine contents in various tissues and plasma

The histamine contents in the various parts of the skin, tissues and plasma were determined after

254 TABLE 2 Histamine contents of lung, intestine, brain, liver, skin of the rats, and plasma after s.c. administration of compound 48/80 (3.0 mg/kg) into the dorsal skin. Tissues were removed 30 min after the s.c. administration of compound 48/80 to determine the histamine contents. Plasma histamine contents were determined at 0.5, 1.0, 2.0 and 3.0 h after the s.c. injection of compound 48/80. Values are means _+S.E.M. The figure in parentheses indicates the number of experiments. Asterisk indicates the statistical difference from control. Tissues or plasma Histamine contents (/~g/g wet weight or ng/ml plasma) Control (saline) Brain Lung Liver Intestine Dorsal skin Abdominal skin Femoral skin Plasma 0.5 h 1.0 h 2.0 h 3.0 h Compound 48/80 0.18_+1.18 (4) 0.18,+ 0.12(5) 2.56_+0.65 (8) 4.15-+ 1.14(40 1.26_+0.13 (10) 1.40-+ 0.18 (8) 17.07_+1.86(10) 16.62_+ 0.72(7) 13.44_+1.18(10) 6.12,+ 0.08(8)** 34.09_+1.18(10) 35.17-+ 4.18(8) 29.14_+4.00(10) 26.23-+ 3.29(8) 25.48 _+3.28 (12) 82.46_+ 14.10 (8) ** 87.06_+ 7.70(4) ** 80.50_+11.00 (4) ** 59.53 _+18.92 (8) * 4. Discussion The s.c. administration of c o m p o u n d 4 8 / 8 0 (3.0 m g / k g ) produced marked increases of PRA, hematocrit and plasma extravasation (figs. 1, 2 and 3). These changes were attenuated or abolished in a dose-related manner by pretreatment with tripelennamine or diphenhydramine, a histamine H I - r e c e p t o r antagonist but not with cimetidine, a histamine H2-receptor antagonist nor with propranolol, a B-adrenoceptor antagonist (figs. 2 and 3). These finding make it possible to speculate that this c o m p o u n d stimulates the cutaneous mast cells and releases histamine, which in turn triggers off plasma extravasation via stimulation of histamine HI-receptors. This then leads to hypovolemia which may directly stimulate the juxtraglomerular cells of the kidney to release renin. This hypothesis would be supported by the following results: (1) the hematocrit increase preceded that of the P R A following s.c. administration of c o m p o u n d 4 8 / 8 0 (fig. 1), (2) the reduction of histamine content as well as the occurrence of plasma extravasation was confined to the site of injection after s.c. administration of c o m p o u n d 4 8 / 8 0 (table 2), (3) s.c. injected c o m p o u n d 4 8 / 8 0 did not cause any significant increase in the nephrectomized rats (table 1). This is consistent with the results of Lembeck and Holzer (1979) who reported that neurogenic plasma extravasation was almost exclusively mediated by the release of a vasoactive substance, probably histamine, from the mast cells and was mediated via the histamine HI-receptor. Further studies are, however, necessary to provide conclusive evidence with respect to the mechanism for the c o m p o u n d 4 8 / 8 0 - i n d u c e d P R A increase, since s.c. administration of histamine did not increase the PRA, and since larger doses (20 or 40 m g / k g i.p.) of either tripelennamine or diphenhydramine did not completely inhibit the increase of P R A caused by c o m p o u n d 4 8 / 8 0 (fig. 2). Lembeck and Holzer (1979) previously reported that the inhibitory effect of mepyramine (10 m g / k g i.p., histamine H I receptor antagonist) plus cimetidine (10 m g / k g i.p.) on the plasma extravasation elicited by either antidromic stimulation of saphenous nerve or infusion of substance P into the femoral artery was nearly 50%. In the present studies tripelennamine

* P < 0.05; ** P < 0.001.

s.c. administration of c o m p o u n d 4 8 / 8 0 (3.0 m g / k g ) into the dorsal skin (table 2). C o m p o u n d 4 8 / 8 0 caused a significant decrease of histamine content only in the injected dorsal skin. N o change of histamine contents could be detected in the lung, intestine, liver, brain and other sites of skin. The plasma histamine content was significantly increased within 30 min after s.c. injection of c o m p o u n d 4 8 / 8 0 (3.0 m g / k g ) . The increase lasted 2 h then started to decline.

3.6. Effect of compound 4 8 / 8 0 on arterial blood pressure in conscious rats

A long-lasting fall in the arterial blood pressure followed the s.c. administration of 0.5 m g / k g isoproterenol. The hypotensive effect was rapid in onset and persited for 90 min. The s.c. injection of c o m p o u n d 4 8 / 8 0 (3.0 m g / k g ) had no effect on the arterial blood pressure of the conscious rats over the 3 h period of observation (data not shown).

255 or d i p h e n h y d r a m i n e injected i.p. in doses of 20 or 40 m g / k g r e d u c e d the c o m p o u n d 4 8 / 8 0 - i n d u c e d P R A increase b y a b o u t 40% or 85%, respectively (fig. 2). It seems likely that access of these inhibitors to the tissue after a b s o r p t i o n into the systemic b l o o d was r a t h e r restricted. A close correlation between h e m a t o c r i t a n d p l a s m a volume has been d e m o n s t r a t e d (Kutscher, 1971), a n d the loss of p l a s m a v o l u m e has been c a l c u l a t e d from the changes in the h e m a t o c r i t (Haefeli a n d Peters, 1971, D u n n et al., 1973). The results of the present e x p e r i m e n t s showing the h e m a t o c r i t increase i n d u c e d b y c o m p o u n d 4 8 / 8 0 in the n e p h r e c t o m i z e d rat without a P R A increase (table 1) i n d i c a t e that the m e c h a n i s m b y which c o m p o u n d 4 8 / 8 0 i n d u c e d h y p o v o l e m i a was n o t relevant to the increase of renin release from the kidney. The s.c. a d m i n i s t r a t i o n of i s o p r o t e r e n o l caused a p r o f o u n d a n d r a p i d fall in arterial b l o o d pressure in the conscious rats b u t s.c. a d m i n i s t r a t i o n of c o m p o u n d 4 8 / 8 0 d i d not affect arterial b l o o d pressure. A relationship b e t w e e n the h y p o t e n s i v e effect a n d P R A increase caused b y i s o p r o t e r e n o l has been well d o c u m e n t e d ( M e y e r et al., 1971; L e e n e n a n d M c D o n a l d , 1974). W e could detect a n e a r l y 4 fold increase of p l a s m a histamine conc e n t r a t i o n 1 h after s.c. a d m i n i s t r a t i o n of 3.0 m g / k g c o m p o u n d 4 8 / 8 0 (table 2). Such an increase of p l a s m a h i s t a m i n e c o n c e n t r a t i o n seems insufficient to decrease arterial b l o o d pressure. F e l d b e r g a n d Talesnik (1953) r e p o r t e d previously that s.c. injection of c o m p o u n d 4 8 / 8 0 (500 /tg, n e a r l y 4.0 m g / k g ) d i d not p r o d u c e general s y m p toms of h i s t a m i n e liberation in c o n t r a s t to i.p. a d m i n i s t r a t i o n of c o m p o u n d 4 8 / 8 0 , which p r o d u c e d signs of severe itching, cyanosis a n d p r o nounced prostration. Asscher, A.W. and S.G. Anson, 1963, A vascular permeability factor of renal origin, Nature 198, 1097. Douglas, W.W., 1980, Histamine and 5-hydroxytryptamine (serotonin) and their antagonists, in: The Pharmacological Basis of Therapeutics (6th ed.), eds. L.A. Goodman and A. Gilman (Macmillan Publishing Co., New York) p. 608. Dunn, F.L., T.J. Brennan, A.E. Nelson and G.L. Robertson, 1973, The role of blood osmolality and volume in regulating vasopressin secretion in the rat, J. Clin. Invest. 52, 3212. Endo, Y., 1981, Simple method for the simultaneous determination of histamine, polyamines and histone H1, J. Chromatog. 205, 155. Erjavec, F., F. Lembeck, T. Florjanc-Irman, G. Skofitsch, J. Donnerer, A. Saria and P. Holzer, 1981, Release of histamine by substance P, Naunyn-Schmiedeb. Arch. Pharmacol. 317, 67. Feldberg, W. and J. Talesnik, 1953, Reduction of tissue histamine by compound 48/80, J. Physiol. (London) 120, 550. Fitzsimons, J.T., 1969, The role of a renal thirst factor in drinking induced by extracellular stimuli, J. Physiol. (London) 201,349. Giese, J., 1963, Pathogenesis of vascular disease caused by acute renal ischaemia, Acta Pathol. 59, 417. Gristwood, R.W., J.C.R. Lincoln, D.A.A. Owen and I.R. Smith, 1981, Histamine release from human right atrium, Br. J. Pharmacol. 74, 7. Haber, E., T. Koerner, L.B. Page, B. Kliman and A. Purnode, 1969, Application of a radioimmunoassay for angiotensin I to the physiologic measurement of plasma renin activity in normal human subjects, J. Clin. Endocrinol. Metab. 29, 1349. Haefeli, L. and G. Peters, 1971, Induction of hypovolaemia by thirst-inducing doses of renin or angiotensin II, Br. J. Pharmacol. 42, 25. Harada, M., T. Takeuchi, T. Fukao and K. Katagiri, 1971, A simple method for the quantitative extraction of dye extravasated into the skin, J. Pharm. Pharmacol. 23, 218. Izumi, H., S.-C. Ho and A.M. Michelakis, 1983a, A comparative study of the effect of cyclocytidine and norepinephrine on renin and esterase release from mouse suhmandibular gland, Naunyn-Schmiedeb. Arch. Pharmacol. 323, 37. Izumi, H., S.-C. Ho, A.M. Michelakis and T. Aoki, 1982, Renin release from mouse submandibular gland in vitro, Endocrinology 111, 758. Izumi, H., S.-C. Ho, A.M. Michelakis and T. Aoki, 1983b, Different effects of compound 48/80 and histamine on plasma renin activity, European J. Pharmacol. 91, 295. Izumi, H., S. Hoshi, S. Mue, T. Takishima, H. Sato and T. Aoki, 1984, The determination of blood histamine in asthmatic patients with a simple and sensitive method, Tohoku J. Exp. Med. 143, 79. Johnson, A.K., J.F.E. Mann, W. Rascher, J.K. Johnson and D. Ganten, 1981, Plasma angiotensin II concentrations and experimentally induced thirst, Am. J. Physiol. 240, R229. Johnson, A.R. and E.G. Erdrs, 1973, Release of histamine from mast cells by vasoactive peptides, Proc. Soc. Exp. Biol. Med. 142, 1252.

Acknowledgements
This work was supported in part by a Grant-in-Aid (59570783) for Scientific Research from the Ministry of Education of Japan (1984).

References
Arvier, P.T., L.A. Chahl and R.J. Ladd, 1977, Modification by capsaicin and compound 48/80 of dye leakage induced by irritants in the rat, Br. J. Pharmacol. 59, 61.

256 Kiernan, J.A., 1975, A pharmacological and histological investigation of the involvement of mast cells in cutaneous axon reflex vasodilatation, Quart. J. Exp. Physiol. 60, 123. Kraly, F.S., 1983, Histamine plays a part in induction of drinking by food intake, Nature 302, 65. Kutscher, C.L., 1968, Plasma volume change during water-deprivation in gerbils, hamsters, guinea-pigs and rats, Comp. Biochem. Physiol. 25, 929. Kutscher, C.L., 1971, Hematocrit, plasma osmolality, and plasma protein concentration as estimators of plasma volume in hooded rats duri.ng food and water deprivation, Physiol. Behav. 7, 283. Leenen, F.H. and R.H. McDonald Jr., 1974, Effect of isoproterenol on blood pressure, plasma renin activity, and water intake in rats, European J. Pharmacol. 26, 129. Lembeck, F. and P. Holzer, 1979, Substance P as neurogenic mediator of antidromic vasodilation and neurogenic plasma extravasation, Naunyn-Schmiedeb. Arch. Pharmacol. 310, 175. Malvin, R.L., D, Mouw and A.J. Vander, 1977, Angiotensin: physiological role in water-deprivation-induced thirst of rats, Science 197, 171. Mann, J.F.E., A.K. Johnson and D. Ganten, 1980, Plasma angiotensin II: dipsogenic levels and angiotensin-generating capacity of renin, Am. J. Physiol. 238, R372. Meyer, D.K., B. Peskar, U. Tauchmann and G. Hertting, 1971, Potentiation and abolition of the increase in plasma renin activity seen after hypotensive drugs in rats, European J. Pharmacol. 16, 278. Paldino, R.L. and C. Hyman, 1954, Mechanism whereby renin increases the rate of T-1824 disappearance from the circulation of rabbits, Proc. Soc. Exp. Biol. Med. 179, 599. Paton, W.D.W., 1951, Compound 48/80: a potent histamine liberator, Br. J. Pharmacol. 6, 499. Vijande, M., M. Gostales and B. Marin, 1978, Sex difference in polyethyleneglycol-induced thirst, Experientia 34, 742. Yamaguchi, K., T. Sakaguchi and K. Kamoi, 1982, Central role of angiotensin in the hyperosmolality- and hypovolaemiainduced vasopressin release in conscious rats, Acta Endocrinol. (Copenh.) 101, 524.