Quantitative immunofluorescence of tyrosine hydroxylase in the adrenal medulla of spontaneously hypertensive rats

The amount of tyrosine hydroxylase protein in the adrenal medulla, which was estimated by a quantitative immunofluorescence method, was higher in spontaneously hypertensive rats than in normotensive control Wistar-Kyoto rats at 4 and 16 weeks of age before and after the development of hypertension.     

Hypothalamic tyrosine hydroxylase activity,plasma gonadotropin and prolactin levels after aminooxyacetic acid in ovariectomized rats

Summary Plasma concentrations of gonadotropin, prolactin and hypothalamic tyrosine hydroxylase (TH) activity were measured in ovariectomized rats treated with aminooxyacetic acid (AOAA), a drug which elevates brain GABA levels. Hypothalamic TH activity was significantly increased with a significant decrease in prolactin (Prl) release. Plasma levels of gonadotropins were not modified by AOAA. These results support an inhibitory action of GABA on Prl release possibly mediated through hypothalamic dopamine.Supported by grants from Indian Council of Medical Research (ICMR), New Delhi. RIA kits for the estimation of LH, FSH and Prl were kindly supplied by Dr A.F. Parlow, NIAMDD-NIH, Bethesda, Maryland, USA. GNB is a UGC research fellow.     

Enhanced reactivity towards flunarizine in cerebrovascular bed of spontaneously hypertensive rats

Summary Cerebral blood flow (CBF) was measured and cerebrovascular resistance (CVR) was calculated in anesthetized spontaneously hypertensive rats (SHR) and normotensive rats (NR) following the administration of incremental dosages of i.v. flunarizine or papaverine. CBF and CVR changes following papaverine were the same in both groups of rats irrespective of the dose of the drug. The effect of flunarizine was much more pronounced in SHR than in NR. The results point out the greater dependency of basal cerebrovascular tone in SHR upon Ca²⁺ influx into vascular smooth muscle cells.     

Development of high blood pressure in spontaneously hypertensive rats is delayed by treatment with cyclosporin at an early age

Summary In spontaneously hypertensive rats the effect of the T-cell inhibitor cyclosporin was studied at different ages. If treatment was started at the age of 2 weeks the development of hypertension was delayed, but the ultimate level of blood pressure was not affected. These results indicate the involvement of immune mechanisms in the early development of hypertension in spontaneously hypertensive rats.     

Increased Na+-H+ exchanger activity in the ileal brush-border membrane of spontaneously hypertensive rats

In the present study, we have examined the intestinal Na⁺ transport, through the Na⁺-H⁺ exchanger, in ileal brush-border membrane vesicles (BBMV) isolated from spontaneously hypertensive rats (SHR), and normotensive Wistar Kyoto (WKY) rats as a control group. Na⁺ uptake into ileal BBMV was stimulated in the presence of a proton gradient (pH 5.5 inside/pH 7.5 outside) in SHR and WKY rats, resulting in a transient accumulation (overshoot) in both groups of rats. No overshoot was observed in the absence of a pH gradient. The magnitude of the accumulation was significantly higher in SHR than in WKY rats. Uptake of Na⁺ at equilibrium was identical in the presence and the absence of a proton gradient and was not changed in SHR. The use of amiloride inhibited pH gradient-driven Na⁺ uptake in a dose-dependent manner with a Ki of 90 μM and 100 μM for SHR and WKY rats, respectively. The relationship between proton gradient-driven Na⁺ uptake and external Na⁺ concentration was saturable and conformed to Michaelis-Menten kinetics in both SHR and WKY rats. Lineweaver-Burk analysis of the pH gradient-driven Na⁺ uptake indicated values of V_max that were significantly increased in SHR compared to WKY rats (11.4±0.55 nmol/mg/8 s vs. 4.96±0.78 nmol/mg/8 s for SHR and WKY rats, respectively). In contrast, similar K_m values for Na⁺ were found between SHR and WKY rats (4.0±0.2 mM vs. 4.9±0.6 mM for SHR and WKY rats, respectively). These studies show derangement in ileal BBMV Na⁺ transport of SHR, which is characterized by increased Na⁺-H⁺ exchanger activity. Received 18 December 1996; received after revision 3 February 1997; accepted 7 February 1997     

Effect of stress on choline acetyltransferase activity of the brain and the adrenal of the rat

Choline acetyltransferase (ChAT) activity was determined in cerebral cortex, hypothalamus, hippocampus, cerebellum, medulla oblongata, midbrain and adrenal gland of rats exposed to acute or chronic stress. The exposure of animals to acute immobilization and cold stress (4°C) for one hour resulted in a significant decline of ChAT activity in all brain regions examined except for the medulla oblongata. Moreover, the exposure to acute stress resulted in significant increase of the same enzyme in the adrenal gland. However, chronic exposure of animals to cold stress (4°C) for 7 days resulted in no significant changes of ChAT activity in all tissues examined except for a decline in the midbrain and an increase in the medulla oblongata. The administration of corticosterone (2.0 mg/kg) 1 h prior to sacrificing caused an effect similar to that of acute stress on ChAT activity in all brain regions except for the hypothalamus and the cerebellum. It was concluded from this experiment that stress-induced changes in the ChAT activity of specific brain regions might be mediated by the adrenal steroids.This work was supported by a grant from the National Aeronautics and Space Administration (NSG 2183 and NAG 2-411), a grant from the National Institutes of Health (NIH RR 0811) and a grant from the Division of Research Resources (NIH grant RR 03020).     

Rate of tryptophan hydroxylation in vivo in brain nuclei of genetically hypertensive rats of the Lyon strain

Summary The rate of tryptophan hydroxylation in vivo is unaltered in brain areas of 5, 9 and 21 week-old Lyon genetically Hypertensive (LH) rats as compared to both Lyon Normotensive (LN) and Low Blood Pressure (LL) rats, except for a decrease in the C1 area of the medulla oblongata in 9 week-old animals.Acknowledgment. The authors wish to thank Dr M.F. Belin, Dr J.F. Pujol and Mrs J. Sacquet for their help during this study. This work was supported by the Fondation pour la Recherche Médicale Française and the C.N.R.S.     

Blood pressure and impairment of endothelium-dependent relaxation in spontaneously hypertensive rats

Summary Correlation between hypertension and impairment of endothelium-dependent relaxation was demonstrated using aortae from certain strains of rats with various levels of spontaneous hypertension. It was also observed that the impairment of endothelium-dependent relaxation is the secondary change due to hypertension, and the level and duration of hypertension is the determinant factor of the impairment.     

Dopamine-beta-hydroxylase activity in adrenal gland and spleen of rats after fasting and cold exposure

Summary Fasting (48 h) results in dopamine-beta-hydroxylase (DBH) release both in adrenal gland and spleen, suggestive of an increase in the activity of these organs. Cold exposure (48 h) produces a dissociation of the, sympathoadrenal response. When both stimuli are simultaneously employed, the DBH response suggests the preponderance of the response to fasting. Plasma DBH is decreased in all groups studied, this could be due to its half-life and the splenic DBH depletion.This work was supported by a grant of Fondo de Investigaciones Sanitarias de la Seguridad Social, No. 83/0905.     

The incorporation of H3-thymidine in the nuclei of the cells of adrenal medulla of rats

Riassunto In questa ricerca si dimostra sintesi di DNA con l'uso di timidina H³ non accompagnata a mitosi nella midollare surrenale di ratto adulto in seguito ad esposizione a bassa temperatura.

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This investigation was supported by a grant to this Department from the Consiglio Nazionale delle Ricerche (No. 04/76/4/3482).     

Selective activation of noradrenergic neurons in the brainstem and spinal cord of young spontaneously hypertensive rats

Summary In young spontaneously hypertensive rats (SHR), dopamine -hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) activities were examined in the brainstem nuclei. Activation of noradrenergic neurons in the locus coeruleus, A2 and spinal intermediolateral cell areas, resulting in enhanced sympathetic nervous activity in the periphery, initiates hypertension. Adrenergic neurons, unchanged in these and A1 cell areas of young SHR, are not involved in the development of hypertension in SHR.     

Features of intracellular calcium distribution in the adipose tissue of spontaneously hypertensive rats (SHR)

The in vitro study of the kinetics of 45Ca efflux from adipose tissue of rats reveals 3 pools of exchangeable calcium. Calcium content in the intracellular pools of adipose tissue of spontaneously hypertensive rats is increased as compared to that in normotensive controls.     

Features on intracellular calcium distribution in the adipose tissue of spontaneously hypertensive rats (SHR)

Summar The in vitro study of the kinetics of⁴⁵Ca effux from adipose tissue of rats reveale 3 pools of exchangeable calcium. Calcium content in the intracellular pools of adipose tissue of spontaneously hypertensive rats is increased as compared to that in normotensive controls.     

Autoradiogaphic investigation of cell proliferation in the adrenal cortex of castrated female rats under the influence of oestradiol

Summary Ovariectomy and subsequent treatment with ovarian hormone produces a temporary increase in DNA-synthesizing cells in the zona glomerulosa of the adrenal cortex.     

T cell and T cell subset determination in normal peripheral blood: Comparison of the indirect immunofluorescence and lymphocytotoxicity techniques

Summary We compared a simple complement-dependent lymphocytotoxicity test with a widely used indirect immunofluorescence procedure to enumerate total T, T helper, and T suppressor lymphocytes in normal blood samples. Results with the two techniques were closely similar.     

Age variation in the increase of hypothalamic and brain stem contents of phenylethanolamine m-octopamine and p-octopamine in spontaneously hypertensive rats (SH Kyoto).

Phenylethanolamine, m-octopamine and p-octopamine contents were determined as a function of age in the hypothalamus and brain stem of spontaneously hypertensive rats and controls Wistar Kyoto. In hypothalamus, the content of the 3 amines was 2--4-fold greater for the SH rats. In the brain stem, the phenylethanolamine and p-octopamine contents were 2--3-fold greater in SH rats but 5--6-fold higher in the case of m-octopamine. The difference appears at 3 weeks and correlates the blood pressure with the increase of age. The significance of these findings is discussed.     

Age variation in the increase of hypothalamic and brain stem contents of phenylethanolamine m-octopamine and p-octopamine in spontaneously hypertensive rats (SH kyoto)

Summary Phenylethanolamine, m=octopamine and p-octopamine contents were determined as a function of age in the hypothalamus and brain stem of spontaneously hypertensive rats and controls Wistar Kyoto. In hypothalamus, the content of the 3 amines was 2–4-fold greater for the SH rats. In the brain stem, the phenylethanolamine and p-octopamine contents were 2–3-fold greater in SH rats but 5–6-fold higher in the case of m-octopamine. The difference appears at 3 weeks and correlates the blood pressure with the increase of age. The significance of these findings is discussed.This work has been completed at the Tokyo University. The author is grateful to Professor Z. Tamura, Dr K. Imai and A. Ooshima as well as to the French Foreign Office and the Japanese Society for the Promotion of Sciences.     

Increased activity of dipeptidyl peptidase IV in serum of hepatoma-bearing rats coincides with the loss of the enzyme from the hepatoma plasma membrane

Summary The specific activity of dipeptidyl peptidase IV (DPPIV E.C. 3.4.14.-) in the plasma membrane of Morris hepatoma 9121 or hepatoma 7777 was 3.5% and 2.9%, respectively, of that in the plasma membrane of rat liver. The enzyme activity in the serum of hepatoma-bearing rats was 141% (hepatoma 91219) and 162% (hepatoma 7777) of the normal value. cytochemical investigation showed that the DPP IV activity was almost completely absent from the hepatoma cell plasma membrane and was not sequestered within these cells. Indirect immunofluorescence staining with a polyclonal antibody directed against DPP IV indicated that the loss of activity was due to the absence of DPP IV molecules in the plasma membrane. The possibility that the enzyme is transferred from the membrane into the serum as a result of structural alterations is discussed.