Pressor response to NaCl solution administered intracerebroventricularly or intracisternally to conscious normotensive and spontaneously hypertensive rats

We examined the central action of NaCl on blood pressure using intracerebroventricular (i.c.v.) and intracisternal (i.c.) injections of hypertonic NaCl solution in conscious, unrestrained spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. The dose-dependent pressor response produced by i.c.v. hypertonic NaCl was greater in SHR than in WKY rats, while the dose-related pressor action produced by i.c. NaCl did not differ between the two strains. The hyperresponsiveness to i.c.v. NaCl in SHR was abolished by pretreatment with an i.c.v. injection of the angiotensin II (ANG II) analogue 1-Sar, 8-IIeu ANG II. Both ANG II and a combination of ANG II and NaCl given by i.c.v. injection had a greater pressor response in SHR than in WKY rats, although both ANG II and phenylephrine given intravenously elevated blood pressure to the same extent in both strains. Furthermore, i.c.v. ANG II both with and without hypertonic NaCl caused dipsogenic behaviour which lasted longer in SHR than in WKY rats. This response to i.c.v. hypertonic NaCl without ANG II was not substantially different between the two strains. Intracisternal hypertonic NaCl did not induce drinking behaviour. These observations suggest that in the SHR, the third ventricle rather than the brain stem is a more sensitive area to NaCl. The brain renin-angiotensin system in the SHR may play an important role in this accelerated pressor response and may be responsible, at least to some extent, for the enhanced reaction to chronic oral salt loading.     

Decreased angiotensin II receptors in subfornical organ of spontaneously hypertensive rats after chronic antihypertensive treatment with enalapril

Angiotensin II (ANG II) receptor density was higher in many brain regions of untreated spontaneously hypertensive rats (SHR) compared to untreated Wistar-Kyoto (WKY) animals. Systemic inhibition of angiotensin converting enzyme with enalapril (25 mg/kg, per os for 14 days) produces a large decrease in ANG II receptors localized exclusively in the subfornical organ (SFO) of the SHR, and no alterations in ANG II receptors in the normotensive WKY rats. Selective decrease of ANG II receptors in the SFO of the genetically hypertensive rats with enalapril may be related to its therapeutic efficacy.     

Differential expression of AT1 receptors in the pituitary and adrenal gland of SHR and WKY

The renin-angiotensin (ANG) system has been implicated in the development of hypertension in spontaneously hypertensive rats (SHR). Because SHR are more susceptible to stress than normotensive Wistar-Kyoto rats (WKY), we measured the mRNA expression of AT1A, AT1B, and AT2 receptors in the hypothalamo-pituitary-adrenal (stress) axis of male SHR in comparison to age-matched WKY at prehypertensive (3 to 4 weeks), developing (7 to 8 weeks), and established (12 to 13 weeks) stages of hypertension. AT1A receptor mRNA was mainly expressed in the hypothalamus and adrenal gland. AT1B receptor mRNA was detected in the pituitary and adrenal gland. AT2 receptor mRNA was prominent only in the adrenal gland. When compared with WKY, SHR showed increased AT1A receptor mRNA levels in the pituitary gland at all ages in contrast to reduced pituitary AT1B receptor mRNA levels. In the adrenal gland of SHR, AT1B receptor mRNA levels were decreased at the hypertensive stages when compared with WKY. The reduced expression of adrenal AT1B receptor mRNA was localized selectively in the zona glomerulosa by in situ hybridization. No differences were observed between WKY and SHR in the expression of hypothalamic ANG receptors. ANG significantly increased plasma levels of adrenocorticotropic hormone (ACTH) and corticosterone in dexamethasone-treated SHR but not in WKY. The aldosterone response to ANG was similar in SHR and WKY. Our results suggest a differential gene expression of AT1A and AT1B receptors in the hypothalamo-pituitary-adrenal axis of SHR and normotensive WKY and imply the participation of AT1 receptors in an exaggerated endocrine stress response of SHR to ANG.     

Vascular angiotensin II receptors in SHR

We investigated the density (Bmax) of angiotensin II (ANG II) receptors in the mesenteric vascular bed of spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto (WKY) control rats. In 12-week-old SHR, the Bmax and the dissociation constant (Kd) of ANG II binding sites were not different from those of WKY rats in the sodium replete state or after sodium depletion. In prehypertensive (4- and 6-week-old) SHR, the Bmax of the vascular ANG II receptors was significantly higher (p less than 0.05) than in age-matched WKY rats. This result could not be attributed entirely to differences in the circulating renin-angiotensin-aldosterone system in 4-week-old-rats. In 6-week-old WKY rats, the plasma renin activity was significantly higher (p less than 0.05), which may account in part for the higher density of ANG II binding sites in SHR. There was an age-related decrease in the number of ANG II receptors in SHR. The increased density of vascular ANG II receptors in young SHR may play a role in the development of high blood pressure in this model of spontaneous hypertension. The higher number of ANG II binding sites in young SHR is not selective for ANG II receptors, since an increased density of alpha 1-adrenergic receptors was also found in the mesenteric arteries of 4-week-old SHR.     

Effects of dietary sodium on brain angiotensin II receptors in spontaneously hypertensive rats

The effects of dietary sodium on the characteristics of angiotensin II (A II) receptor sites in the hypothalamus-thalamus-septum-midbrain (HTSM) region were examined in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Twenty-four SHR and 24 WKY were divided into two groups respectively, which were maintained on high sodium diets or low sodium diets for 4 weeks, respectively. The binding capacity and affinity of the A II receptors were measured by radioreceptor assay. In WKY, the binding capacity of the A II receptors in the high sodium group was significantly lower than that in the low sodium group. On the other hand, the binding capacity of A II receptors in the high sodium group was significantly lower than that in the low sodium group. On the other hand, the binding capacity of A II receptors was not significantly different between high and low sodium groups in SHR. The secretion of arginine vasopressin (AVP) increased significantly in SHR with high sodium intake. The present results suggest that in WKY the decrease of the binding capacity of the A II receptors in the HTSM region in response to a high sodium intake serves to attenuate an osmotical stimulus to AVP secretion. However, in SHR such a regulatory mechanism as adjusting the binding capacity of the A II receptors is lacking, and this seems to be responsible, at least in part, for the enhanced secretion of AVP on the sodium loading.     

Increased concentration of angiotensin II binding sites in selected brain areas of spontaneously hypertensive rats

We studied the density of the angiotensin II (Ang II) binding site in discrete brain nuclei of 4-week-old and 14-week-old spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar-Kyoto (WKY) control rats by autoradiographic binding techniques. Tissue sections were incubated in vitro with 3 nmol/l [125I]Sar1Ang and results were analysed by computerized microdensitometry and by comparison with 125I-standards. Both young and adult SHR (aged 4 and 14 weeks, respectively) had significantly higher Ang II binding site concentrations in the median preoptic nucleus (MPO), subfornical organ (SFO), paraventricular nucleus (PVN) and nucleus of the solitary tract (NTS) when compared to age-matched WKY control rats. No significant difference was found between strains in other brain areas such as the olfactory bulb, suprachiasmatic nucleus (SCh), inferior olive (IO) and area postrema (AP). It was observed that the concentration of Ang II binding sites increased with age in PVN of both SHR and WKY, while the number of binding sites in the MPO and IO decreased with age. In SHR, alteration in Ang II binding is restricted to brain nuclei involved in the central pressor action of Ang II and seems to be related to the development and maintenance of spontaneous hypertension.     

Effect of low doses of angiotensin II perfusion on the hypotensive action of captopril in anaesthetized normotensive and spontaneously hypertensive rats

The effect of intravenous (i.v.) captopril on mean arterial blood pressure (MABP) of anaesthetized normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats perfused i.v. with two doses of angiotensin II (ANG II; 2.9 and 5.8 pmol/kg per min) was studied to determine the role of the suppression of plasma ANG II in the hypotensive action of captopril. The reduction of MABP by captopril was attenuated in WKY and abolished in SHR by the highest dose of ANG II; it was unchanged in WKY and attenuated in SHR by the lowest dose of ANG II. The suppression of plasma ANG II thus explains a minor part of the acute reduction of MABP by captopril in WKY and a major part of this action in SHR. Plasma ANG II contributes to the maintenance of high blood pressure in SHR.     

Abnormal regulation of adrenal angiotensin II receptors in spontaneously hypertensive rats

The aldosterone response to angiotensin II is blunted in spontaneously hypertensive rats (SHR). To determine whether this blunting is due to a defect in angiotensin II receptors, we assessed angiotensin II binding to intact adrenal glomerulosa cells in SHR and normotensive Wistar-Kyoto rats (WKY) that had been fed high or low sodium diets before sacrifice. In rats on high salt intake, we observed no difference between the two strains in either receptor affinity (Kd = 1.0-1.2 nM) or binding capacity (36,000-38,000 receptors/cell). When sodium-restricted, WKY increased receptor content more than fourfold to 167,000 sites/cell. SHR increased receptor number to only 103,000 sites/cell, which was significantly (p less than 0.01) less than the WKY increase. The cause of the abnormal receptor regulation remains unclear. Two known receptor regulators, the plasma angiotensin II level and the state of potassium balance, were similar in the two strains. Our results suggest that the blunted aldosterone response to angiotensin previously reported in SHR is due to abnormal angiotensin receptor up-regulation in the adrenal gland in response to sodium restriction.     

Glomerular dimensions in spontaneously hypertensive rats: effects of AT1 antagonism

OBJECTIVE: A reduction in glomerular number and/or size has been implicated in the development of hypertension. This study investigated whether differences in glomerular number and/or size occur during the development of hypertension in the spontaneously hypertensive rat (SHR) and whether angiotensin II is responsible for any glomerular differences. METHODS: SHR (n=6) and Wistar-Kyoto (WKY) rats (n=6) were administered the angiotensin II type I receptor antagonist TCV-116 from 4 to 10 weeks of age. At 10 weeks of age, the kidneys from these rats and those from untreated SHR (n=6) and WKY rats (n=6) controls were perfusion fixed at physiological pressures and analysed using unbiased stereological techniques. RESULTS: There were no significant differences in glomerular number, glomerular volume or total glomerular volume between SHR and WKY rats. Treatment of SHR with TCV-116 significantly lowered systolic blood pressure but had no significant effect on glomerular number or volume or total glomerular volume. Treatment of WKY rats with TCV-116 reduced systolic blood pressure, body weight, glomerular volume and total glomerular volume; however, total glomerular volume per body weight of treated WKY rats was not significantly different from that of untreated WKY rats. CONCLUSION: There were no differences in glomerular number or volume in SHR compared with WKY rats at 10 weeks of age. We therefore conclude that glomerular changes are not responsible for the development of hypertension in SHR. Angiotensin II, via the type 1 receptor, does not contribute to glomerular growth during the development of hypertension in the SHR.     

Levels of Immunoreactive Angiotensin II in Microdissected Nuclei from Adult wky and Sh Rat Brain

Levels of immunoreactive angiotensin II (ANG II) were measured by radioimmunoassay in microdissected nuclei from the brain of the spontaneously hypertensive rat (SHR) and its normotensive control, the Wistar Kyoto rat (WKY). The nuclei assayed included the paraventricular nucleus of the hypothalamus (PVH), locus coeruleus (LC), nucleus of the solitary tract (NTS), dorsal motor nucleus of the vagus nerve (DMN of X) and the Al region of the medulla. Sections of cerebral and cerebellar cortex were used as controls. Levels of immunoreactive ANG II ranged from 0.24 nmoles/g protein to 0.93 nmoles/g protein, with SHR brain containing significantly higher levels than WKY brain in the PVH and NTS. higher levels of immunoreactive ANG I1 than WKY brain, the difference was not significant. the A1 region between the two species, and no detectable ANG I1 immunoreactivity was found in cerebral or cerebellar cortex. Since each of the nuclei studied has been implicated in the neural control of cardiovascular function, the increased levels of immunoreactive ANG I1 in the SH rat brain nuclei indicates a possible role for ANG I 1 in the pathophysiology of hypertension in the SHR model. Although LC and DMN of X in the SHR brain contained No significant difference was found in     

Angiotensin II binding sites in the superior cervical ganglia of spontaneously hypertensive and Wistar-Kyoto rats after preganglionic denervation

In the superior cervical ganglia (SCG) binding site density of angiotensin II (ANG II) was higher in adult spontaneously hypertensive rats (SHR) (571 +/- 29 fmol/mg protein) compared to that in the adult Wistar-Kyoto rat (WKY) (375 +/- 9 fmol/mg protein, P less than 0.05). The ANG II binding density was significantly decreased in the SCG of SHR (-59%, P less than 0.01) and of WKY (-39%, P less than 0.05) after unilateral preganglionic denervation (operated v sham-operated ganglia). Part of the binding sites in the superior cervical ganglia may be present in or be associated to preganglionic nerves, and the number of these sites is higher in SHR.     

Cerebral capillary bed structure of normotensive and chronically hypertensive rats

In this study cerebral capillary bed structure and the effects of chronic hypertension on these systems have been assessed in 6- to 7-month-old spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. Capillary diameter (D), profile frequency (Na), volume fraction (Vv), and surface area (Sv) were quantitated by light microscopic morphometry of eight brain areas including the sensorimotor cortex and subfornical organ. Previously presented data from normotensive Sprague-Dawley rats (SpD) of similar age were also compared. Within each of the three rat strains, D, Na, Vv, and Sv varied among brain areas. For the sensorimotor cortex and subfornical organ, capillary profile frequency differed significantly among the three rat strains. In SHR and WKY, there was an inverse correlation between profile frequency and diameter, i.e., as Na increased among brain areas, D decreased. In six brain areas capillary volume fraction and surface area were identical in SHR and WKY, but were lower in SpD. Consistent differences between SHR and WKY were found only for the subfornical organ, which suggests some involvement of this structure in hypertension. Since there were few statistically significant differences between SHR and WKY and many statistically significant differences between the two normotensive strains, cerebral capillary bed structure seems to be independent of arterial blood pressure in most brain areas of these rats.     

1,2-Diacylglycerol content in myocardium from spontaneously hypertensive rats during the development of hypertension

Summary 1,2-Diacylglycerol (DAG) has been considered to play an important role as an activator of protein kinase C in the signal transduction of inositol phospholipid metabolism. To examine the relation of 1,2-DAG in heart tissues to cardiac hypertrophy associated with hypertension, we measured the amount of 1,2-DAG in spontaneously hypertensive rat (SHR) hearts at 4,10 and 20 weeks of age, and in age-matched normotensive Wistar-Kyoto (WKY) rat hearts using thin-layer chromatography with flame ionization detection (TLC-FID). Significant cardiac hypertrophy was found in 4-week-old SHR, while SHR did not yet have significant hypertension. Major phospholipids such as phosphatidylcholine and phosphatidylethanolamine increased from 4 to 20 weeks in the myocardium, but there was no difference between the two strains. The cholesterol levels of 4- and 20-week-old SHR were significantly higher than WKY rats. The 1,2-DAG contents of SHR hearts were significantly higher than WKY rats at 4 weeks. An increase in the RNA contents of SHR hearts were significantly higher than WKY rats at 4 weeks. An increase in the RNA content was also observed in 4-week-old SHR hearts. However, analysis of the fatty acid composition of 1,2-DAG revealed no difference between the two strains. However, there was no significant difference in the 1,2-DAG content or in its fatty acid composition between SHR and WKY rat hearts at 10 and 20 weeks of age. It is suggested that an increase in the 1,2-DAG content of SHR hearts during the early stages appears related to the initiation of cardiac hypertrophy in SHR hearts before developed hypertension.     

Impaired prostaglandin E(2)/prostaglandin I(2) receptor-G(s) protein interactions in isolated renal resistance arterioles of spontaneously hypertensive rats

The protective effect of vasodilator agents linked to the cAMP pathway is less effective for buffering the vasoconstrictor effect of angiotensin II in young animals with genetic hypertension. To determine the underlying cellular mechanism, experiments were performed on freshly isolated preglomerular resistance arterioles obtained from kidneys of 7-week-old spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). Specific high-affinity saturable binding of (3)H-prostaglandin (PG) E(2) revealed 1 receptor class in renal microvessels; PGE(2) receptor density was similar in SHR and WKY (106 versus 115 fmol/mg; P>0.8), as was receptor affinity (3.6 versus 3.5 nmol/L; P>0.7). Basal cAMP activity was similar in renal arterioles from SHR and WKY. A major finding was that PGE(2), PGI(2), and isoproterenol produced weaker stimulation of cAMP formation in arteriolar cells of SHR (P<0.02). In contrast, GTPgammas and forskolin stimulated cAMP generation to a similar degree in both rat strains, which suggests normal adenylate cyclase activity in hypertension-prone SHR. Immunoblots revealed the presence of 3 classes of G proteins (G(s), G(i), and G(q)) in preglomerular arterioles. The relative amounts of discernible G-protein alpha-subunits in renal resistance vessels did not differ between SHR and WKY. These results extend previous in vivo studies of abnormal renal vascular reactivity in SHR and more directly localize defective coupling of the prostaglandin and beta-adrenergic receptors to a stimulatory G protein and cAMP production in freshly isolated preglomerular arteriolar cells of young SHR. This dysfunction may be due to an abnormal interaction between prostaglandin receptors and G(s) protein that leads to inefficient coupling of initiating steps in the cAMP-protein kinase A cascade during the development of hypertension.     

Body weight of the spontaneously hypertensive rat during the suckling and weanling periods

The body weight of the spontaneously hypertensive rat (SHR) during the suckling, birth to 3 weeks of age, and weanling period, through 6 weeks of age, is compared to that of its normotensive controls, the Wistar Kyoto (WKY) and Wistar (WR) rats. Litters were normalized to 6 pups per dam just after birth. The WR is larger than the SHR and WKY at all time points examined. The WKY is similar in weight to the SHR at birth but larger than the SHR at the other time points studied. These findings suggest the WR is inherently a larger rat than the SHR and WKY during the suckling and weanling periods while the development of a weight difference between the SHR and WKY after birth suggest an extrauterine influence.     

Increased Atrial Natriuretic Factor Receptor Density in Cultured Vascular Smooth Muscle Cells of the Spontaneously Hypertensive Rat: Revised

To explore the role of the atrial natriuretic factor (ANF) system in the pathophysiology of hypertension we examined the binding kinetics of synthetic ANF to cultured vascular smooth muscle cells (VSMCs) derived from the spontaneously hypertensive rat (SHR) and two normotensive controls-the Wistar Kyoto (WKY) and American Wistar (W). The number of maximal binding sites (Bmax) per cell (mean ± SEM; × 10³) were: SHR = 278.0 ± 33.0, WKY = 28.3 ± 7.1 and W = 26.6 ± 4.2. The differences between the SHR and normotensive strains were significant at p<0.001. The equilibrium dissociation constant (Kd; × 10^?9M) was higher in SHR VSMCs (0.94 ± 0.14) than in WKY (0.22 ± 0.09; p <0.01) and W (0.39 ± 0.14; p <0.02) cells. The plasma levels of the imnunoreactive ANF were higher in SHR than the normotensive controls. We suggest that the relatively greater ANF receptor density in cultured VSMCs of the SHR represents a response to the invitro environment which is relatively more deficient in ANF for VSMCs of the SHR as compared with the normotensive rats. Thus, the capacity of the SHR VSMC to regulate ANF receptor density appears to be independent of the blood pressure level.     

Effect of Sodium Chloride on Opiate Receptor Binding in Spontaneously Hypertensive Rats: Dependence on Age

The effect of 100 mM sodium chloride on the binding of [³H]-naltrcxone to rat brain opiate receptors in spontaneously hypertensive (SHR) and normotensive (WKY) rats was studied. The percentage increase in binding in the presence of sodium chloride did not vary with age in WKY rats. Brain homogenates from 4 week old SHR rats incubated with 100 mM sodium chloride exhibited a similar increase in binding compared to age matched WKY rats. In contrast, brain preparations from 6, 14 and 20 week old SHR rats were more sensitive to sodium chloride, and the increase in binding of [³H]-naltrcxone was significantly greater in these animals than in corresponding normotensive ones. Since blood pressure is increased in SHR rats compared to WKY rats at these ages, these results suggest that elevated blood pressure may be correlated with an increase in opiate receptor sensitivity to sodium chloride. The effect of In vivo sodium chloride was examined by feeding the animals a diet containing 4% salt. This concentration of salt did not significantly alter the binding of [³H]-naltrcxone to rat brain homogenates prepared from 8 week old SHR rats. These results suggest that higher levels of sodium chloride and longer exposure to the diet may be required to observe the salt sensitivity produced by 100 mM salt in the In vitro radioreceptor assay.     

Cerebral glucose utilization and blood flow in adult spontaneously hypertensive rats.

Not only blood pressure but also behavioral activity, brain morphology, and cerebral ventricular size differ between young spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. This suggests that cerebral blood flow and cerebral metabolism may vary between these two rat strains. To test this hypothesis, we measured local cerebral glucose utilization in 31 brain areas of 26-30-week-old rats. Local cerebral blood flow was also assessed in these same areas. Cerebral glucose utilization was measured by the 2-deoxyglucose method; cerebral blood flow was determined by the iodoantipyrene method. In virtually all gray matter structures, the apparent rate of glucose utilization was lower in SHR than in normotensive WKY rats; the interstrain differences varied significantly among structures and were statistically significant (uncorrected t tests) in 14 of 28 gray matter areas. Local cerebral blood flow was fairly similar in the two rat strains. The coupling of blood flow to glucose utilization varied significantly among brain areas in normotensive WKY rats as well as in SHR. In a number of gray matter structures, the coupling of flow to metabolism differed between hypertensive and normotensive animals. These data suggest that for many brain areas, either glucose utilization or glucose partitioning differs between WKY rats and SHR.     

1,2-diacylglycerol content in thoracic aorta of spontaneously hypertensive rats

Phosphoinositide metabolism participates in the control of cell calcium homeostasis. Because a notable neutral lipid (1,2-diacylglycerol) is generated from phosphoinositide hydrolysis and is assumed to be a secondary messenger, we determined 1,2-diacylglycerol content and its fatty acid profiles in the thoracic aorta of spontaneously hypertensive rats (SHR) and compared it with those of normotensive Wistar-Kyoto (WKY) rats. After the aorta was exposed to 10(-5) M norepinephrine as a stimulant, 1,2-diacylglycerol content in SHR was significantly higher by 33% than in WKY rats at 4 weeks of age, whereas there was no difference in 1,2-diacylglycerol content between the two strains at 20 weeks of age. Before norepinephrine stimulation, there was no significant difference in 1,2-diacylglycerol level between the two strains at 4 weeks of age. Analysis on a gas chromatograph showed that 1,2-diacylglycerol was composed of similar molecular species of fatty acids in aortas obtained from SHR and WKY rats. On the other hand, the cholesterol content of aortas was higher in SHR than in WKY rats at 20 weeks of age, whereas the difference at 4 weeks was not significant. Phosphatidylcholine, phosphatidylethanolamine, and triglyceride showed no significant difference between the two strains. It is concluded that norepinephrine-induced 1,2-diacylglycerol production increases in the thoracic aorta of SHR before the development of hypertension.