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E Dux

Publications and source records attributed to E Dux.

At least 37 records · Page 2Linked to original sources

Reverse pinocytosis induced in cerebral endothelial cells by injection of histamine into the cerebral ventricle.

Histamine dihydrochloride (10 micrograms of 500 micrograms/ml) was infused during 1 min into the lateral cerebral ventricle of rats, which resulted in a significant stimulation of pinocytosis in the endothelial cells. Systemic injections of mepyramine or metiamide could not prevent this activation. In contrast, ranitidine, injected with histamine was able to inhibit the stimulation of pinocytosis. Albumin exudation from the blood was not found. There was also no change in water and electrolyte contents of the brain tissue. The results suggest that histamine reaching the abluminal membrane can activate the pinocytosis in the cerebral endothelial cells in the reverse direction, i.e., from brain to blood, without opening the blood-brain barrier.

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Kinetics of protein phosphorylation in microvessels isolated from rat brain: modulation by second messengers.

The role of second messengers in the regulation of protein phosphorylation was studied in microvessels isolated from rat cerebral cortex. The phosphoproteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the kinetics of 32P incorporation into specific protein substrates were evaluated by computer-aided x-ray film densitometry. With the use of this method, Ca2+-calmodulin (CAM)-, Ca2+/phospholipid (PK C)-, cyclic GMP (cGMP)-, and cyclic AMP (cAMP)-dependent protein kinases were detected. CAM-dependent protein kinase proved to be the major phosphorylating enzyme in the microvascular fraction of the rat cerebral cortex; the activity of cGMP-dependent protein kinase was much higher than that of the cAMP-dependent one. Autophosphorylation of both the alpha- and beta-subunits of CAM-dependent protein kinase and the proteolytic fragment of the PK C enzyme was also detected. The kinetics of phosphorylation of the individual polypeptides indicate the presence in the cerebral endothelium of phosphoprotein phosphatases. The phosphorylation of proteins in the cerebral capillaries was more or less reversible; the addition of second messengers initiated a very rapid increase in 32P incorporation, followed by a slow decrease. Because the intracellular signal transducers like Ca2+ and cyclic nucleotides are frequently regulated by different vasoactive substances in the endothelial cells, the modified phosphorylation evoked by these second messengers may be related in vivo to certain changes in the transport processes of the blood-brain barrier.

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Calcium in the mitochondria following brief ischemia of gerbil brain.

Calcium was localized in neurons and glial cells of the CA1 region of hippocampus and in neurons of parietal cortex with the oxalate-pyroantimonate electron cytochemical method following recovery from 5-min ischemia of gerbil brain. During the early postischemic recirculation phase a maximum amount of calcium deposits was detected in mitochondria of all investigated cells. A second rise in mitochondrial calcium deposits was observed in parallel with the onset of ultrastructural degeneration of mitochondria and the loss of cellular integrity beginning 6 h postischemia.

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Protective effect of antihistamines on cerebral oedema induced by experimental pneumothorax in newborn piglets.

As a consequence of general hypoxaemia evoked experimentally by bilateral pneumothorax, brain oedema of vasogenic type developed in newborn piglets after 4 h survival. Histamine receptor antagonists, mepyramine (H1-receptor blocker), metiamide, cimetidine and ranitidine (H2-receptor antagonists) were administered either intraperitoneally or intrathecally to check to what extent the formation of brain oedema could be reduced. Mepyramine and ranitidine decreased the accumulation of water, sodium and albumin in the parietal cortex. By measuring the concentration of histamine, the presence of a histamine pool was demonstrated in the cerebral microvessels. The results suggest that histamine, if released upon hypoxic injury from the microvascular store, can take an important part in the development of vasogenic brain oedema.

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Calcium in hippocampus following lidocaine induced seizures: an electron cytochemical study.

The effect of lidocaine seizures on cellular accumulation of calcium was studied in hippocampal subfields CA1 and CA3 and the dentate gyrus of rats, using the combined oxalate-pyroantimonate method. The specificity of the reaction was ascertained by EGTA treatment and X-ray microanalysis. In control rats, calcium was visualized between myelin lamellae of axons, in synaptic vesicles and in some lysosomes. Two hours after onset of lidocaine seizures selective neuronal degenerations appeared in hippocampal subfields CA1 and CA3 but not in the dentate gyrus. Calcium deposits were present in numerous mitochondria of pyramidal cells and, occasionally, also of neuroglial cells. Many of these mitochondria exhibited ultrastructural alterations. Calcium uptake was most prominent in the CA3 sector but was also present in the CA1 subfield as well as the dentate gyrus. Intracellular calcium uptake, in consequence, is not the unique attribute of selectively vulnerable hippocampal neurons.

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The blood-brain barrier in hypoxia: ultrastructural aspects and adenylate cyclase activity of brain capillaries.

The ultrastructure of brain microvessels, their permeability to serum albumin, the activities of some endothelial enzymes and the effect of histamine were investigated in rats after a prolonged hypobaric-hypoxic treatment. After prolonged hypoxia, the permeation of serum albumin into endothelial cells increased together with the number of pinocytotic vesicles of the endothelium. Intracarotid histamine stimulated this process even further, and its effect was mediated by H2-histamine receptors. After hypoxia the specific activity of capillary alkaline phosphatase and gamma-glutamyl transpeptidase remained unchanged, while that of adenylate cyclase was greatly increased. Histamine did not modify the structure of tight junctions of isolated capillaries of normoxic animals. Both hypoxia- and histamine-induced modification of the brain microvessels were accompanied by an increase of pinocytosis, which may be stimulated by the activation of capillary adenylate cyclase.

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Regulation of the macromolecular transport in the brain microvessels: the role of cyclic GMP.

The possible effects of dibutyryl cyclic GMP (db-cGMP), the lipid-soluble derivative of cyclic GMP, on brain microvessels were studied by light microscopic detection of albumin and quantitative electron microscopy. Different concentrations (25, 50, 100 and 200 micrograms) of db-cGMP were given to adult rats by intracarotid infusion, while in the controls, the animals were infused either with Krebs-Ringer solution or with the same solution containing butyrate or cyclic GMP. In contrast to the controls, db-cGMP was found to be able to increase the permeability of brain microvessels to albumin in a dose-dependent manner. At the same time, the number of transport vesicles being indicative of the pinocytotic activity of the endothelial cells was also increased. These results indicate that, similarly to the cyclic AMP whose effects have been revealed earlier, the cyclic GMP-system may also be involved in the regulation of opening mechanisms of the blood-brain barrier.

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Effects of histamine on brain capillaries. Fine structural and immunohistochemical studies after intracarotid infusion.

The effect of histamine administered via the common carotid artery on the transport processes of brain capillaries was investigated in rats. The fine structure of endothelial cells and the glial end-feet system was studied by electron microscopy and the serum albumin was visualized for light microscopy by the peroxidase-antiperoxidase (PAP) immunohisto-chemical reaction. Sixty microgram per milliliter histamine enhanced the penetration of serum albumin into the capillaries while the number of pinocytotic and coated vesicles significantly increased in the capillary endothelium. Oedematous swelling of the glial end-feet system was also observed. The stimulatory effect of histamine on the transcapillary transport could not be inhibited by a histamine H1-receptor antagonist, mepyramine. By contrast, metiamide, a histamine H2-receptor antagonist prevented both ultrastructural changes and albumin penetration in the brain capillaries to occur.

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Histamine-stimulated prostaglandin synthesis in rat brain microvessels.

The histamine-sensitivity of prostacyclin and prostaglandin synthesis was investigated in isolated brain microvessels prepared from normal and hypoxic exercised rats. 10(-4) M histamine stimulated the in vitro synthesis of all components of arachidonate cascade. The chronic hypoxic exercise also resulted in an enhanced production of each fraction. Hypoxia and histamine showed an additive effect in the synthesis of PGE2 only. The possible molecular mechanism induced by hypoxia and histamine is discussed.

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Prostacyclin and prostaglandin synthesis in isolated brain capillaries.

The synthesis of prostacyclin and prostaglandins was examined in isolated blood-free brain capillaries of guinea-pigs and rats using 1-14C-arachidonic acid as a precursor. The main prostaglandins synthesized by guinea-pig microvessels were prostaglandin D2 and prostaglandin E2. Substantially less prostaglandin F2 alpha or the prostacyclin stable metabolite, 6-oxo-prostaglandin F1 alpha was synthesized. Rat capillary prostaglandin distribution differed substantially from that of the guinea-pigs although the principle prostaglandin was also PGD2. Total prostaglandin conversion was greater in guinea-pig capillaries than in the rat. Norepinephrine stimulated the prostaglandin forming capacity of blood free cerebral microvasculature of guinea-pigs. Prostacyclin and prostaglandins could be involved in the activity dependent regulation of regional cerebral blood flow and permeability.

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Further data on the androgenic dependency of the skeletal musculature: the effect of prepubertal castration on the structural development of the skeletal muscles.

The effect of prepubertal castration was investigated in the soleus, semimembranosus and levator ani muscles of rat, using light and electronmicroscopic morphometric methods. After castration the semimembranosus and the levator ani muscles showed significant morphometrical alterations. The weight of the muscles diminished, the diameter of the white fibers decreased. The ultrastructure of these muscles showed a well expressed myofibrillar atrophy. The changes in the semimembranosus muscle were less severe. The slow oxidative soleus muscle did not show any similar alterations. The changes in the levator ani and semimembranosus muscles could be prevented or moderated by testosterone substitution.

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Microvessels from the spinal cord: isolation procedure and characterization of the fraction.

A simple method for the preparation of microvessels from the spinal cord is described. The fraction being enriched in the microvessels is characterized by morphological techniques and compared biochemically with the original suspension. The method makes possible detailed studies on the metabolism of microvessels and can be complementary to procedures that have been developed for separating the capillaries from other brain tissues.

Alkaline Phosphatase↗

Histamine in brain capillaries.

A high amount of histamine was found in capillaries isolated by subcellular fractionation from the brain. In view of the important effects of histamine on vascular permeability in peripheral vessels, it was thought that histamine had a similar function in the cerebral vasculature. Intracarotic histamine infusion resulted in an enhanced pinocytosis of endothelial cells and te oedematous swelling of the astrocytic end-feet system. [3H]-Histamine, injected in the cerebral ventricles, accumulated in the capillary wall. Histamine and cimetidine activated hydroxyfatty acid and prostaglandin D2 synthesis in isolated brain capillaries. The possible function of the capillary histamine in the regulation of permeability of microvessels is discussed.

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Prostacyclin and prostaglandin synthesis in rat brain microvessels: characteristic changes in hypoxaemia.

The effects of hypoxaemia on the synthesis of prostacyclin and prostaglandins in the cerebral microvessels and on the ultrastructure of brain cortex capillaries were investigated in three groups of rats. Series I: rat embryos were kept in hypoxic-hypobaric condition for 42 hours. It resulted in the enhancement of synthesis of PG2alfa and PGE2 even two months after birth. The fine structure of the capillary endothelium remained without any pathological change. Series II: adult male rats were on hypoxic-hypobaric environment for 42 hours. There was a slight increase in the synthesis of PGE2 and PGD2 while the ultrastructure of brain capillaries was found to be normal. Series III: the hypoxic-hypobaric condition lasted for 240 hours in adult male rats. This long term hypoxaemia affected greatly the prostaglandin synthesis of brain cortex capillaries and resulted in changes in the fine structure of brain microvessels as well. The possible molecular mechanism activated by hypoxaemia in brain capillaries is discussed.

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