Brain Of Spontaneously Hypertensive Rats Research Articles

Effects of ethanol on brain monoamine content of spontaneously hypertensive rats (SHR).

Spontaneously hypertensive rats (SHR) were administered either 2.4 g/kg ethanol or an isocaloric glucose daily for 4 weeks and the levels of norepinephrine (NE), epinephrine (EP), dopamine (DA), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in different brain regions were determined. Results indicated a 3-fold increase in NE level in brain stem and hypothalamus and more than 2-fold increase in DA in corpus striatum in alcohol-treated rats as compared to controls. There was a significant increase in the level of DA in the corpus striatum but the levels in cerebral cortex, brain stem and hippocampus were decreased instead. Decreases in 5-HT levels were found in hypothalamus, brain stem, cortex and cerebellum of alcohol-treated brain as compared to untreated controls. These results indicate alterations of the biogenic amine contents in different regions of the SHR brain after chronic ethanol ingestion. Since stimulated release of biogenic amines in the SHR brain has been implicated in the regulation of blood pressure, changes due to ethanol ingestion may be a risk factor in hypertensive patients.

Parenchymal microvascular systems and cerebral atrophy in spontaneously hypertensive rats

Spontaneously hypertensive rats (SHR) are hypertensive, hyperactive, and hydrocephalic; furthermore SHR have smaller brain volume and weight than age-matched, normotensive Wistar-Kyoto rats (WKY). At 6–7 months of age, local cerebral glucose is sizably lower in SHR than WKY. The hypothesis that these several abnormalities of SHR lead to variations in cerebral microvascular bed morphology was tested in 6–7-month-old SHR and WKY by quantitating various parameters of small, intermediate, and large parenchymal microvessels (grouped by luminal diameter) in 21 brain areas. Within each rat strain, the microvascular bed properties such as vessel profile frequency (density) varied considerably among the 21 brain areas. In opposition to the hypothesis, mean luminal diameter as well as profile frequency, surface area, and luminal volume of the microvascular beds per unit tissue mass were virtually identical in each brain area of SHR and WKY for the three groups of microvessels. These findings coupled with the reports of less tissue per structure but similar density of neurons throughout the brain of SHR and WKY indicate that there are fewer neurons and less vascular tissue per brain structure in 6–7-month-old SHR than WKY; in addition, they suggest a linkage between the size of parenchymal microvascular beds and the surrounding nervous tissue.

Increased angiotensin II type-1 receptor gene expression in neuronal cultures from spontaneously hypertensive rats.

In this study we compared the expression of angiotensin II type 1 (AT1) receptor messenger RNA (mRNA) and AT1 receptors in neurons cultured from Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rat brains. Neuronal cultures from the hypothalamus and brain-stem of 1-day-old SH rats exhibited approximately 4-fold higher steady-state levels of AT1 receptor mRNA than the corresponding WKY cultures. This was attributable to greater levels of both AT1A and AT1B receptor mRNA subtypes in SH rat neuronal cultures compared with WKY rat neurons. SH rat neuronal cultures also exhibited increased numbers (approximately 2.3-fold) of binding sites for [3H]DuP753, an AT1 receptor selective ligand, and enhanced (approximately 3.4-fold) stimulation of inositol phospholipid hydrolysis by angiotensin II compared with WKY neurons. By contrast, cultured astroglia from SH and WKY rat brain exhibited no significant differences in either the levels of AT1 receptor mRNA or the specific binding of [3H]DuP753. These data suggest that in SH rat neurons, AT1 receptor transcription and translation is increased, compared with neurons from WKY rats.

Neuroanatomical differentiation in the brain of the spontaneously hypertensive rat (SHR). I. Volumetric comparisons with WKY control.

A series of measurements was made to assess the morphology of the brain of the spontaneously hypertensive rat (SHR). The SHR brain was smaller than that of age-matched normotensive Wistar-Kyoto (WKY) controls in a majority of measures of external surface landmarks. This reduction in size was evident in the youngest age group examined (94 days) and persisted in older groups (170, 240 and 350 days). The brain of the SHR was also smaller in terms of brain weight and brain weight:body weight ratios. Section-by-section digitized analyses of coronal histologic sections from 94-day-old rats revealed significant reductions in mean cross-sectional area and volume of midbrain/pons (10%) and hindbrain (11%) regions, but not of forebrain, in the SHR. Alterations in the mediolateral dimension, particularly within the pontomedullary brainstem, accounted for more of these volumetric changes than those in the dorsoventral dimension. Using the same coronal sections, it was found that surface areas and volumes of five individual nuclei/fiber tracts, selected for their involvement in central cardiovascular regulation, were significantly decreased in the SHR. The largest reduction in volume (30%) was found in the nucleus tractus solitarius, the primary site of termination of afferent baroreceptor fibers. No differences in surface area or volume were found in that portion of the cerebroventricular system (aqueduct of Sylvius) associated with the periventricular grey region, or in the inferior colliculus, which is not thought to be involved in cardiovascular control. These observations not only have practical implications, but suggest that the pathophysiological condition expressed as spontaneous hypertension in this widely-used model may be related to morphological alterations in the central nervous system.

Smaller local brain volumes and cerebral atrophy in spontaneously hypertensive rats.

Spontaneously hypertensive rats (SHR) have enlarged cerebral ventricles from 8 weeks of age onward and smaller brains than age-matched, normotensive Wistar-Kyoto (WKY) rats (controls). At 6-7 months of age local cerebral glucose utilization is apparently lower in many brain areas of SHR relative to WKY rats. These observations led to the hypothesis that there are morphological differences between these two strains of rats in many, if not all, brain areas. This hypothesis was tested in 6-7-month-old SHR and WKY rats by quantitating 1) the volumes of the ventricular system, whole brain, six gray matter structures, and two white matter areas; 2) the thickness of two regions of the cerebral cortex; and 3) the frequency of neuronal nuclei (neuronal frequency) in nine brain areas. Ventricular volume was twofold greater in SHR than in control rats. The volumes of the entire brain and all six gray matter structures plus the thickness of the two cortical regions were 11-25% less in SHR. Neuronal frequency was, however, similar in the two rat strains. The latter finding coupled with the smaller regional tissue volumes indicates appreciably fewer neurons per brain structure in young adult SHR than in controls. These results indicate significant cerebral structural differences between young adult SHR and WKY rats and suggest that structure as well as metabolism are abnormal in the SHR brain.

Effects of gamma-vinyl GABA (vigabatrin) on blood pressure and body weight of hypertensive and normotensive rats.

Inactivation of GABA was inhibited by gamma-vinyl GABA (GVG) and the effects of the increased GABA level in the brain on blood pressure and body weight of spontaneously hypertensive rats (SHR) and normotensive rats (WKY) were investigated. When started at the age of 8 weeks or 5 weeks, treatment of SHR and WKY with GVG (150 mg/kg, s.c.) for several weeks did not influence systolic blood pressure. In 1-week old SHR, treatment with GVG (up to 150 mg/kg, s.c.) abolished the rise in blood pressure until animals were 8 weeks old. Thereafter, arterial blood pressure started to increase but it remained distinctly lower than that in untreated animals. When started at the age of 1 week, treatment with GVG for 7 weeks did not influence arterial blood pressure in WKY. GVG delayed increase in body weight in SHR and WKY, irrespective of their age. GVG greatly increased GABA levels in the hypothalamus, frontal cortex, brainstem and rest of the brain in both WKY and SHR. It is concluded that an increase in the GABA level in the brain leads to a delay in the development of hypertension in young SHR. Hence, development of genetic hypertension seems to be susceptible to activation of the GABAergic system in a very early critical phase only.

Brain dopamine D-2 receptor mechanisms in spontaneously hypertensive rats

Brain dopaminergic function was studied in spontaneously hypertensive rats (SHR) with the selective dopamine D-2 antagonist sulpiride. Sulpiride dose-dependently inhibited locomotor activity of normotensive Wistar-Kyoto rats (WKY). SHR showed an increase in locomotor activity in response to low doses of sulpiride, whereas no effect was observed of higher doses. In a two-bottle salt-preference test, WKY showed increased preference for an 0.9% saline solution after treatment with sulpiride, whereas total fluid intake remained the same. In SHR, sulpiride influenced neither saltpreference nor total fluid intake. SHR and WKY with a unilateral lesion of the median forebrain bundle showed similar turning behaviour in response to treatment with amphetamine. Pretreatment with 100 mg/kg sulpiride virtually abolished amphetamine-induced turning in WKY, but had little effect in SHR. Sulpiride dose-dependently increased serum prolactin concentrations in WKY and SHR. However, the increase was significantly greater in SHR. Dopamine D-2 receptor binding was measured with in vitro autoradiography, using [ 125I]-sulpiride as the ligand. Binding density was similar in the caudate nucleus and substantia nigra of SHR and WKY brain. Concentrations of the dopamine metabolites DOPAC and HVA, but not of dopamine itself, were significantly increased in frontal cortex, striatum and hypothalamus after treatment with 100 mg/kg sulpiride. There were no significant differences between SHR and WKY in the increase in the DOPAC/DA and HVA/DA ratio. These data show that SHR show differential changes in their response to central dopamine D-2 blockade when compared to WKY. Thus, in some tests (locomotor activity after high doses, salt preference, turning behaviour), SHR respond less to sulpiride. In contrast, in other aspects (locomotor activity after low doses, serum prolactin levels), SHR show increased functional responses to sulpiride when compared to WKY. These differences are not the result of changes in postsynaptic receptor density, as dopamine D-2 receptor binding and the increase in dopamine turnover after D-2 receptor blockade were normal in SHR. It is concluded that SHR show increased functional in vivo central dopamine neurotransmission, possibly due to higher stress-induced central dopamine release and/or an altered coupling of dopamine D-2 receptors to (one of) their transduction mechanisms or second messenger systems. This could explain the relative lack of postsynaptic receptor blockade by sulpiride in this strain. However, presynaptic receptor density or sensitivity is upregulated in SHR to counteract the higher dopaminergic activity, which might explain the disinhibitory effect of low doses of sulpiride found in this strain.

Repeated Idazoxan Increases Brain Imidazoline Receptors in Normotensive (WKY) but Not in Hypertensive (SHR) Rats

The specific binding of [3H]idazoxan in the presence of 10(-6) M (-)-adrenaline was used to evaluate the density of imidazoline receptors in the brain of spontaneously hypertensive (SHR) rats and sex- and age-matched normotensive Wistar-Kyoto (WKY) rats. In SHR rats the density of imidazoline receptors (cerebral cortex, hypothalamus, and medulla oblongata) was not different from that in normotensive (WKY) rats. However, repeated treatment with idazoxan consistently increased (23-80%) the density of imidazoline receptors in the various brain regions of WKY rats but not in SHR rats. In normotensive Sprague-Dawley rats, repeated treatment with the imidazoline drugs idazoxan and cirazoline also increased (33-37%) the density of imidazoline receptors in the cerebral cortex. The lack of regulation by idazoxan of the density of imidazoline receptors in the brain of SHR rats might reflect the existence of a relevant abnormality of these receptors in this genetic model of hypertension.

Effects of chemical stimulation and lesion of the rostral ventrolateral medulla in spontaneously hypertensive rats

We investigated whether or not the rostral ventrolateral medulla (RVL) participates in maintaining a higher arterial pressure of spontaneously hypertensive rats (SHR). SHR and Wistar-Kyoto rats (WKY) were anesthetized with chloralose, paralyzed with tubocurarine, and artificially ventilated. Chemical excitation of the RVL neurons by microinjection of excitatory amino acids ( l-glutamate) elicited a dose-dependent pressor response in both the SHR and WKY groups. The magnitude of increase in the arterial pressure (AP) was similar in both groups, but the present increase of AP was higher in WKY than in SHR. Chemical lesions of the bilateral RVL produced by the microinjection of tetrodotoxin decreased the AP to the spinal level, which was similar in both groups. The results indicate that the RVL functions as the sole sympathetic output of the brain in SHR, and suggest that the RVL neurons may be tonically overactive to maintain hypertension in SHR.

Reduced calcium sensitivity of dihydropyridine binding to calcium channels in spontaneously hypertensive rats.

To explore the role of calcium channels in hypertension, dihydropyridine ([3H]PN200-110) binding to heart, brain, and skeletal muscle microsomes of 4-, 8- and 15-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats was measured. At a constant Ca2+ ion concentration (pCa 3.0), maximal binding (Bmax) of dihydropyridine binding to heart and brain microsomes was significantly enhanced in 8- and 15-week-old SHR compared with WKY rats (p less than 0.01), whereas this phenomenon was not observed in 4-week-old SHR and WKY rats. Bmax and dissociation constant (Kd) values for skeletal muscle microsomes from SHR showed no difference compared with WKY rats irrespective of age. Dihydropyridine binding to heart microsomes, brain microsomes, and solubilized skeletal muscle microsomes exhibited strong calcium dependence. The Ca2(+)-dependent dihydropyridine binding curves for heart showed a Hill slope, and pK 0.5 values for 15-week-old SHR and WKY rats were 0.70 +/- 0.12 and 4.66 +/- 0.12 versus 0.72 +/- 0.12 and 5.66 +/- 0.08 (n = 4, mean +/- SD), respectively, indicating that 15-week-old SHR require 10-fold higher calcium concentration than WKY rats to promote dihydropyridine binding. The pK 0.5 values of calcium for brain and solubilized skeletal muscle calcium channels in 15-week-old SHR were also significantly lower than in WKY rats. This difference first became apparent in SHR and WKY rats as early as 4 and 8 weeks after birth. These results suggest that enhancement of calcium channel density might occur in the heart and brain of SHR in response to elevated blood pressure and that reduced calcium sensitivity of dihydropyridine binding to calcium channels might be a primary characteristic of this rat strain.

Hindbrain Ischemia Produced by Bilateral Vertebral Artery Occlusion and Moderate Hypotension in Spontaneously Hypertensive Rats

Hindbrain ischemia was induced by bilateral vertebral artery occlusion and moderate hypotension in spontaneously hypertensive rats (SHRs). Mean arterial blood pressure was lowered to 80 mmHg in SHRs and to 50 mmHg in Wistar-Kyoto rats (WKYs) by a controlled hemorrhage, and then the vertebral artery was bilaterally occluded through alar foramina of the first cervical vertebra. Following vertebral occlusion, blood flow of the cerebellum was significantly decreased to 9.4 +/- 2.0 mL/100g/min (+/- SEM) while flow of the cerebrum remained at 32.1 +/- 5.4 in SHRs. In contrast, cerebellar blood flow in WKYs was preserved at 24.2 +/- 2.9 mL/100g/min. Brain lactate, pyruvate, and adenosine triphosphate (ATP) were determined in SHRs after sixty minutes of hypotension with or without vertebral occlusion. Although infratentorial metabolites were actually unaltered in rats with hypotension alone, infratentorial lactate and lactate/pyruvate ratio significantly increased to 14.38 +/- 3.61 mmol/kg and 67.7 +/- 12.1, respectively, with a concomitant decrease in ATP in SHRs with hypotension and vertebral occlusion. Bilateral vertebral artery occlusion, together with moderate hypotension, was shown to produce a marked reduction of cerebellar blood flow and to induce ischemic metabolic changes in the infratentorial brain in SHRs.

Comparison of angiotensin metabolism by brain membranes from SHR and WKY rats

The ability of membrane-associated peptidases from the brains of spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats to metabolize iodinated angiotensin ( 125I-Ang II) and 125I-Ang III was compared. 125I-Ang II was metabolized to 125I-Ang III and other fragments exclusively by membrane-associated peptidases. In contrast to 125I-Ang III which was effectively degradated by both membrane-associated and residual cytosolic peptidases, 125I-Ang II was unaltered by contanimating cytosolic enzymes. The ability of SHR-derived membranes to metabolize 125I-Ang II and produce 125I-Ang III was enhanced when compared to membranes from WKY rats. No difference was observed in the ability of membrane or cytosolic enzymes from SHR and WKY rats to degrade 125I-Ang III. These data are consistent with an increased availability of Ang III in the brains of SHRs.

Immunoreactive atrial natriuretic peptide in neuronal and glial cells of spontaneously hypertensive rat brain

Neuronal and glial cultures from the hypothalamic-brain stem areas of 1-day-old normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rat brains stained positively with atrial natriuretic peptide (ANP)-specific antibodies. The endogenous levels of the ANP immunoreactivity in WKY neuronal and glial cultures were 17.0 +/- 2.2 and 14.3 +/- 2.7 pg/mg, respectively. Comparable neuronal and glial cultures from SH rat brains contained a 48 to 70% decrease in the endogenous ANP immunoreactivity levels. Culture media from both brain cell types also contained ANP immunoreactivity, the levels of which are significantly higher than those found in the cells. However, similar to endogenous levels, the media levels of immunoreactive ANP in SH neuronal and glial cultures were significantly reduced compared with WKY brain cultures. These observations demonstrate that endogenous ANP-like immunoreactivity is found in neuronal and glial cells and is released into the media. The levels of peptide are reduced in cultures of SH compared with WKY cultures, suggesting a genetically controlled difference between the hypertensive and normotensive rat strains long before hypertension develops.

Brain synaptosomal Ca 2+ uptake: Comparison of Sprague-Dawley, Wistar-Kyoto and spontaneously hypertensive rats

1. 1. K +-stimulated 45Ca 2+ uptake by synaptosomes was measured with respect to the strain differences between Sprague-Dawley (SD), Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). 2. 2. 45Ca 2+ uptake by synaptosomes isolated from cerebral cortex of SD, WKY and SHR was measured at 15, 30, 60, 120 and 240 sec time periods. 3. 3. The sequence of both the magnitude and rate of resting and depolarization-dependent 45Ca 2+ uptake was SHR > WKY > SD. 4. 4. The fastest rates of resting and depolarization-dependent 45Ca 2+ uptake occurred in each rat during the first 15 sec and uptake rates dropped off quickly in both resting and depolarization states. 5. 5. At 15 sec, there were significant differences between SHR and WKY, while there were no significant differences between WKY and SD. 6. 6. The results suggest that an important alteration in Ca 2+ channel characteristics may occur in SHR brain synaptosomes.

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 A1 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. Although LC and DMN of X in the SHR brain contained higher levels of immunoreactive ANG II than WKY brain, the difference was not significant. No significant difference was found in the A1 region between the two species, and no detectable ANG II 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 II in the SH rat brain nuclei indicates a possible role for ANG II in the pathophysiology of hypertension in the SHR model.

Lack of alpha-1-adrenergic receptor-mediated downregulation of angiotensin II receptors in neuronal cultures from spontaneously hypertensive rat brain

Neuronal cells from Wistar Kyoto (WKY) and spontaneously hypertensive (SH) rat brains were established in culture to compare the expression of angiotensin II (Ang II) specific receptors and their regulation by norepinephrine (NE). Neurons from SH rat brains possess twice more Ang II specific receptors and expressed a proportional increase in Ang II stimulated [3H]-NE uptake compared with WKY neurons. NE caused a dose-dependent decrease in 125I-Ang II binding in WKY neurons, an effect not observed when neurons from SH rat brains were incubated with NE. These observations suggest that the lack of NE-induced downregulation of Ang II receptors in neuronal cultures is genetically regulated.

Ganglionic, spinal cord and hypothalamic atrial natriuretic factor

We report the occurrence of the atrial natriuretic factor (ANF) prohormone in the hypothalamus, spinal cord and sympathetic ganglia determined by measurement of immunoreactive ANF by two peptide-specific radio-immunoassays with antibodies against near-C-terminal and near-N-terminal portions of ANF prohormone. This suggests local ANF generation in neural structures. In spontaneously hypertensive rats (SHR) we found an elevated ANF-C content in all tissues along the pathway of increased efferent sympathetic outflow, which is present in this animal model. The ANF-N was augmented in SHR only in the hypothalamus. This indicates an overall increase of neural ANF in SHR. The reported neuroinhibitory function of increased neural ANF, however, was attenuated by a decrease in the number of some brain and peripheral ganglionic ANF binding sites in SHR. It remains to be determined whether the increased neural ANF in SHR is a primary phenomenon or a compensatory increase induced by high blood pressure.

Brain angiotensin in the developing spontaneously hypertensive rat.

There are several factors in the manifestation of high blood pressure in spontaneously hypertensive rats (SHR) which implicate a central role for brain angiotensin II (Ang II). We have measured levels of angiotensin in the brain of SHR and rats of the Wistar-Kyoto strain (WKY). The experiments were carried out in 2-, 4-, 14- and 20-week-old rats. Areas of brain from rats were homogenized and purified with SepPak C-18 cartridges. The levels were measured by radio-immunoassay whose detection limit was 1.95 pg/tube. Significant differences were found between the different age groups and between SHR and controls. In the hypothalamus, there was a consistent elevation of brain Ang II in SHR as compared to WKY in all age groups. Cerebellum also had higher levels in SHR, especially in rats at 2 and 4 weeks of age. Brainstem levels were significantly higher in SHR only in the 14-week-old age group. Plasma levels during these time periods did not differ significantly between the strains. The results demonstrate changes in brain Ang II with development. At an early age, there are high levels of Ang II in the hypothalamus and cerebellum which do not correlate with hypertension but may be important for the development of hypertension. The higher levels of brain Ang II in SHR support the hypothesis that hypertension in SHR is related to brain Ang II activity.

Increased expression of α1-adrenergic receptors in the hypothalamus of spontaneously hypertensive rats

The specificity and molecular weights of α 1-adrenergic receptors in various tissues of spontaneously hypertensive (SH) rat were compared with normotensive controls (Wistatt-Kyoto; WKY) with the use of [ 125I]HEAT and [ 125I]azidoprazosin, specific α 1-adrenergic receptor antagonists. Binding of [ 125I]HEAT to membranes prepared from SH rat brain hypothalamus was significantly higher. due to a 75% increase in the B max, than the WKY control. In contrast, the B max and K d of [ 125I]HEAT binding to brainstem and liver membranes from SH rats were not significantly different from those of WKY controls. Competition-inhibition data suggested similar pharmacological specificity with potencies in the order of prazosin > yohimbine > propranolol for both WKY and SH rat membranes prepared from liver, hypothalamus, brainstem and neuronal cultures. Photoaffinity labeling of α 1-adrenergic receptors from hypothalamus, brainstem and neuronal cultures using [ 125I]azidoprazosin followed by SDS-PAGE and autoradiography showed the presence of one major band with a molecular weight (MW) of 105,000 Da for both WKY and SH rats. In contrast, labeling of liver α 1-adrenergic receptors revealed one major band with a MW of 60,000 Da. Quantitation of the 105,000-Da band from SH rat hypothamic membranes demonstrated a 52% higher intensity compared with WKY controls. Neuronal cultures prepared from 1-day-old SH rats showed a similarly greater intensity of the 105,000-Da band compared with WKY controls. These observations indicate that: (1) the MW of α 1-adrenergic receptors in the hypothalamus, brainstem and neuronal culture is significantly higher compared with the liver receptor: and (2) increased expression of the α 1-adrenergic receptors in the SH rat is specific for the hypothalamus and a similar increase is maintained in neuronal cultures prepared from neonatal rat hypothalamus-braistem. These observations suggest that increased α 1-adrenergic receptor expression may be the result of their increased synthesis or decreased degradation. In addition, the results suggest that the increase in α 1-adrenergic receptors may be related to the genetic expression of hypertension, since the same property is observed in neuronal cultures from 1-day-old SH rats, where blood pressure is not yet elevated. In addition, these data also suggest that the central α 1-adrenergic receptor, although pharmacologically similar to the peripheral receptor, may be structurally unique.

α1‐Adrenergic Receptors in Neuronal Cultures from Rat Brain: Increased Expression in the Spontaneously Hypertensive Rat

Neuronal cells in primary culture from 1-day-old brains of normotensive, Wistar-Kyoto strain (WKY) and spontaneously hypertensive (SH) rats have been utilized to study the expression of alpha 1-adrenergic receptors. Binding of a selective alpha 1 antagonist, [125I]2-[beta-(4-hydroxy-3-iodophenyl)-ethylaminomethyl]-tetralone ([125I]HEAT) to neuronal membranes prepared from primary brain cultures of WKY and SH rats was 75-80% specific, rapid, and time-dependent although the binding was 1.5-2 times higher in neuronal membranes from SH rat brain cultures. Kinetic analysis of the association and dissociation data demonstrated no significant differences between rat strains. Competition-inhibition experiments provided IC50 values for various antagonists and agonists in the following order: prazosin less than phentolamine less than yohimbine less than phenylephrine less than norepinephrine less than propranolol, suggesting that [125I]HEAT bound selectively to alpha 1-adrenergic receptors. Scatchard analysis of the binding data provided straight lines for both strains of rats, indicating the presence of a homogeneous population of binding sites. It also showed that the increase in the binding in neuronal cells from SH rat brains over those from normotensive WKY controls was a result of an increase in the number of alpha 1-adrenergic receptors. Incubation of neuronal cultures from both strains of rats with phenylephrine, an alpha 1-adrenergic agonist, caused a time- and dose-dependent decrease in the binding of [125I]HEAT. This decrease was due to a decrease in the number of alpha 1-adrenergic receptors.(ABSTRACT TRUNCATED AT 250 WORDS)