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Time course and disposition of fucose radioactivity in rat hippocampus. A biochemical and microautoradiographic study.

Male adult rats were injected intraventricularly with L-[1-3H]fucose. At various intervals, ranging between 30 min and 11 days, one-half of the brain was prepared for microautoradiography, and the hippocampus from the other side was prepared for biochemical investigations. The TCA-precipitable proteins from the hippocampus homogenate were maximally labeled at between 8 and 24 h and remained at a high radioactivity level even 11 days after [3H]-fucose injection, the labeling being predominantly present in the solubilized insoluble proteins. Using gel electrophoretic separation, study of Tris-soluble material indicated a rapid turnover of soluble fucose-containing glycoproteins, whereas several slow-migrating bands of solubulized proteins revealed a time course suggesting the presence of fucose-containing glycoproteins with slower turnover rates. Using microautoradiography, a rapid labeling of neuronal cell bodies of the hippocampus was found, whereas the nuclei were not labeled. Perikarya were maximally labeled 4 h after [3H]fucose application. The radioactive material was continuously transported from the soma into the corresponding fiber layers, the latter being maximally labeled at a pulse interval of one day; even 10 days later a considerable amount of radioactivity could be detected in the neuropil.

Animals↗

Effect of hypothalamic lesions on interaction of centrally administered ANF and the circulating sodium-pump inhibitor.

We investigated the possible central interaction of atrial natriuretic factor (ANF) and an endogenous Na+, K(+)-ATPase (Na-pump) inhibitor in normal rats. Release of an endogenous Na-pump inhibitor associated with deoxycorticosterone acetate-salt hypertension may be regulated in the anteroventral third ventricle (AV3V) area of the CNS. We reported earlier that bolus injection of synthetic 26-amino acid ANF (Arg101-Tyr126, 6 micrograms/250 g rat) into the lateral brain ventricle (ICV) promotes the appearance in the plasma of a Na-pump inhibitor in rats. To determine whether the AV3V area of the brain is involved in the ICV effect of ANF, we introduced electrolytic lesions in this area. This treatment abolished the appearance of the Na-pump inhibitor after intraventricular injection of ANF. To further localize the area and the pathways involved in the interaction of ANF and the Na-pump inhibitor, we produced bilateral medial coronal knife cuts designed to transect the medially coursing pathway through the periventricular tissue of the AV3V region between the level of the medial preoptic area and the anterior hypothalamic nuclei. These knife cuts also abolished the appearance of the Na-pump inhibitor after ICV injection of ANF. Our data to date indicate that centrally administered ANF promotes the appearance of a Na-pump inhibitor in the plasma. A central site of interaction between ANF and the Na-pump inhibitor appears to be the AV3V area and a medial pathway coursing caudally from the AV3V region.

Analysis of Variance↗

[Effect of intracerebroventricular injection of caerulein on stress-induced gastric mucosal lesion in rats].

The effect of caerulein injected into cerebroventricle on the stress-induced gastric mucosal lesion of rats with tied four limbs immersed in water was investigated. Caerulein (1.0 ng/rat) reduced significantly gastric mucosal lesion, decreased gastric acid content, but increased the gastric content of the mucus and PGE2. Under electromicroscope, morphological sign of hyposecretion in the parietal cells and hypersecretion in the mucus cells could be seen. The effect of caerulein could be prevented by intraventricular injection of naloxone or subcutaneous injection of indomethacin, but not by atropine, phentolamine and propranolol. The results indicate that the protective effect of caerulein on the gastric mucosa is mediated partly by central morphine receptors and partly by enhancement of endogenous PGE2.

Animals↗

Effects of intracerebroventricular administration of prostaglandin D2 and angiotensin II on blood pressure in conscious rats.

To evaluate the role of brain prostaglandins in the regulation of blood pressure, we examined the effects of intracerebroventricular injections of prostaglandin D2, angiotensin II and indomethacin on blood pressure in conscious rats. Intraventricular administration of prostaglandin D2, the major prostaglandin synthesized in the rat brain, did not elicit a significant change in blood pressure. On the other hand, intraventricular injection of angiotensin II resulted in an increase in blood pressure in a dose-related manner. However, this central pressor effect of angiotensin II was not affected by intraventricular pretreatment with indomethacin. Indomethacin per se did not induce any change in blood pressure. These results suggest that prostaglandin D2 in the brain does not play an important role in the regulation of blood pressure in conscious rats. It is also suggested that the central pressor effect of angiotensin II is not mediated by prostaglandin biosynthesis in the central nervous system.

Angiotensin II↗

Effect of beta-adrenergic drugs on LH, FSH, and growth hormone (GH) secretion in conscious, ovariectomized rats.

The effects of third ventricular (3V) injection of the beta-adrenergic antagonist, propranolol (PROPR), a selective beta 1-antagonist, metoprolol (MET), a selective beta 2-antagonist, IPS 339, and a beta-adrenergic agonist (-) isoproterenol (ISOPR), on plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), and growth hormone (GH) were studied in conscious, ovariectomized (OVX) rats. Samples were removed from unrestrained rats which had been previously implanted with atrial and 3V cannulae, and plasma hormone levels were determined by radioimmunoassay (RIA). Intraventricular injection of PROPR (30 micrograms), MET (40 micrograms), or IPS 339 (20 micrograms) induced a gradual elevation in plasma GH concentrations, whereas ISOPR (30 micrograms) reduced plasma GH. ISOPR (30 micrograms) brought about a decrease in plasma LH concentrations, but PROPR, MET and IPS 339 had no effect on LH levels. PROPR (30 micrograms) increased plasma FSH concentrations, but there was no significant effect of MET, IPS 339 or ISOPR on FSH secretion. The results indicate that the beta-adrenergic system can inhibit the release of GH, LH, and FSH. This system appears to have a tonic inhibitory effect on GH and FSH but not LH release in the OVX rat.

Adrenergic beta-Agonists↗

Lentiviral vectors mediate efficient and stable gene transfer in adult neural stem cells in vivo.

Modulation of adult neurogenesis may offer new therapeutic strategies for various brain disorders. In the adult mammalian brain the subventricular zone (SVZ) of the lateral ventricle is a region of continuous neurogenesis. Lentiviral vectors stably integrate into dividing and nondividing cells, in contrast to retroviral vectors, which integrate only into dividing cells. We compared their potential for gene transfer into both quiescent and slowly dividing stem cells as well as into more rapidly dividing progenitor cells. In contrast to retroviral vectors, stereotactic injection of lentiviral vectors into the SVZ of adult mice resulted in efficient and long-term marker gene expression in cells with characteristics of both immature type B cells and migrating precursor cells. After migration along the rostral migratory stream and differentiation, the number of enhanced green fluorescent protein (eGFP)-expressing granular and periglomerular interneurons increased over time in the ipsilateral olfactory bulb. Moreover, the number of eGFP-labeled neuronal progenitor cells in the SVZ increased over time. By intraventricular injection of lentiviral vectors we could restrict gene transfer to ependymal cells and type B astroglial-like stem cells. In conclusion, lentiviral vectors surpass retroviral vectors in efficient long-term in vivo marking of subventricular zone stem cells for basic research and therapeutic applications.

Animals↗

Transduction of the choroid plexus and ependyma in neonatal mouse brain by vesicular stomatitis virus glycoprotein-pseudotyped lentivirus and adeno-associated virus type 5 vectors.

Evaluation of gene transfer into the developing mouse brain has shown that when adeno-associated virus serotype 1 (AAV1) or AAV2 vectors are injected into the cerebral lateral ventricles at birth, widespread parenchymal transduction occurs. Lentiviral vectors have not been tested by this route. In this study, we found that injection of lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein (VSV-G) resulted in targeted transduction of the ependymal cells lining the ventricular system and the choroid plexus along the entire rostrocaudal axis of the brain, whereas a Mokola pseudotype transduced only a few cells after injection into the neonatal ventricle. In contrast, when lentiviral vectors pseudotyped with either VSV-G or Mokola glycoprotein are injected into the adult mouse brain, they transduce similar patterns of cells. An Ebola-Zaire-pseudotyped vector did not transduce any neonatal CNS cells, as was also the case for adult parenchymal injections. Long-term gene expression (12 months) occurred with a constitutively active mammalian promoter and a self-inactivating long terminal repeat (LTR), whereas the cytomegalovirus promoter in a vector with an intact LTR was expressed only in short-term experiments. We found that an AAV5 vector also targeted the ependymal and choroid plexus cells throughout the ventricular system. This vector exhibited limited penetration from the ventricle to other structures, which was significantly different from the previously reported patterns of transduction after intraventricular injection of AAV1 and AAV2 vectors.

Animals↗

Effects of acute central and peripheral administration of nicotine on ascending dopamine pathways in the male rat brain. Evidence for nicotine induced increases of dopamine turnover in various telencephalic dopamine nerve terminal systems.

The actions of intraventicular injections and intravenous infusions of nicotine were studied on dopamine stores and turnover in discrete areas of the forebrain of normal male rats. This was done by measuring the decline of the dopamine stores after tyrosine hydroxylase inhibition using alpha-methyl-tyrosine methyl ester (H44/68). The dopamine concentrations in the various telencephalic dopamine nerve terminal systems were measured using the Falck-Hillarp methodology involving quantitative microfluorimetry. The catecholamine concentrations in the anteromedial frontal cortex were measured biochemically using high pressure liquid chromatography combined with electrochemical detection. Intraventricular experiments. The dopamine levels in discrete areas of nuc. caudatus and nuc. accumbens were significantly reduced even with the lowest dose of nicotine (1 microgram/rat). Intraventricular injections of nicotine in a dose of 100 microgram/rat produced significant increases of dopamine turnover in various types of dopamine nerve terminal systems in the nuc. caudatus, nuc. accumbens and tuberculum olfactorium, and following a dose of 10 microgram/rat increases of dopamine turnover were observed in the medial part of the nuc. caudatus. Furthermore, nicotine (100 microgram/rat) significantly increased noradrenaline but not dopamine turnover within the anterofrontal cortex. Intravenous experiments. The dopamine levels were selectively reduced by nicotine (1000 microgram/kg) in the cholecystokinin positive and negative dopamine nerve terminal systems of the nuc. accumbens. On the other hand, dopamine levels in the anteromedial frontal cortex were increased after this dose of nicotine. Intravenous infusions of nicotine (10-1000 microgram/kg) produced dose-related increases of dopamine turnover in the various dopamine nerve terminal systems analysed in the telencephalon. These effects became significant with a dose of 1000 microgram/kg/h. The dopamine terminals in the nuc. caudatus showed a higher sensitivity to intravenous infusions of nicotine, being affected by 10-100 microgram/kg of nicotine. These findings suggest that relatively low dose of nicotine via an activation of central nicotine-like cholinergic receptors can reduce dopamine concentration and increase dopamine turnover in discrete limbic and striatal areas. These actions may in part represent the neurochemical basis for the rewarding actions of nicotine and for nicotine dependence in man.

Afferent Pathways↗

Effect of blockade of dopaminergic receptors on acetylcholine (Ach)-induced alterations of plasma gonadotropin and prolactin (Prl) in conscious ovariectomized rats.

The participation of acetylcholine (Ach) in the control of gonadotropin and prolactin (Prl) release was evaluated in intact or pimozide-treated conscious, ovariectomized (OVX) rats bearing indwelling 3rd ventricular and jugular venous cannulae by measuring plasma hormone levels by radioimmunoassay (RIA). Third ventricular injection of 20 or 100 microgram of atropine sulfate (2 microliter) significantly lowered plasma LH and Prl levels. Intraventricular injection of 20 microgram Ach (2 microliter) induced a significant elevation in plasma LH and suppression of Prl within 5 min of injection, whereas FSH was elevated only at 30 min. Intraventricular atropine blocked the action of Ach to elevate FSH and LH and to lower Prl. Intravenous pulse injection of atropine at a dose of 5 mg/kg suppressed plasma LH and Prl. Blockade of dopamine (DA) receptors with pimozide prevented the FSH- and LH-releasing and Prl-lowering effects of intraventricular Ach. The atropine-induced suppression of LH release was reversed and an elevation occurred in animals which were pretreated with the DA receptor blocker. Atropine injection had no effect on already elevated Prl levels in pimozide-pretreated animals. The results provide further evidence for cholinergic stimulation of gonadotropin and inhibition of Prl release and indicate that cholinergic control may be mediated via the tuberoinfundibular dopaminergic neurons.

Acetylcholine↗

Variability of bregma in 300 gram Long-Evans and Sprague-Dawley rats.

Bregma is used as an anatomical reference point for most freehand and some stereotaxic injections into the rat brain. The variability of bregma with respect to the intraaural line has been examined previously for rats of a wide range of weights. We examined this variability in 282-326 gram male rats of the Long-Evans and Sprague-Dawley strains. In both strains, mean bregma was less than that stated in a widely used stereotaxic atlas. Furthermore, the mean bregma of the Sprague-Dawley rats was significantly greater than that for the Long-Evans strain. The results suggest that bregma is inadequate as an anatomical reference for intraventricular injections in rats unless the volume injected is a substantial fraction of ventricular volume.

Animals↗

Cytosine arabinoside cerebrospinal fluid kinetics.

To better characterize the disposition of cytosine arabinoside (Ara-C) in cerebrospinal fluid (CSF), its kinetics were studied in seven patients with meningeal leukemia in complete remission. After intraventricular injection of 30 mg Ara-C, CSF and plasma samples were obtained over a 24-hr period. Ara-C levels were measured by a reverse-phase HPLC assay (with a sensitivity of 0.5 microM in CSF and 1.0 microM in plasma) that readily separated Ara-C from its major metabolite uracil arabinoside (Ara-U). Elimination of Ara-C from CSF followed a biphasic pattern, with an initial t1/2 of 1 hr and a terminal t1/2 of 3.4 hr. Ara-C clearance from CSF was 0.42 ml/min, suggesting that drug elimination was primarily by CSF bulk flow. The ratio of the AUC of Ara-U to the AUC of Ara-C was 0.08, indicating only minor metabolism of Ara-C to Ara-U in CSF, in contrast to that after systemic Ara-C. Despite initial CSF Ara-C concentrations exceeding 2 mM, Ara-C was not detectable in plasma in any patient. Intraventricular Ara-C results in very high levels in CSF, but systemic tissues are relatively spared from exposure to Ara-C.

Adolescent↗

Degeneration of hippocampal CA3 pyramidal cells induced by intraventricular kainic acid.

Degeneration of hippocampal CA3 pyramidal cells was investigated by light and electron microscopy after intraventricular injection of the potent convulsant, kainic acid. Electron microscopy revealed evidence of pyramidal cell degeneration within one hour. The earliest degenerative changes were confined to the cell body and proximal dendritic shafts. These included an increased incidence of lysosomal structures, deformation of the perikaryal and nuclear outlines, some increase in background electron density, and dilation of the cisternae of the endoplasmic reticulum accompanied by detachment of polyribosomes. Within the next few hours the pyramidal cells atrophied and became electron dense. Then these cells became electron lucent once more as ribosomes disappeared and their membranes and organelles broke up and disintegrated. Light microscopic changes correlated with these ultrastructural observations. The dendritic spines and the initial portion of the dendritic shaft became electron dense within four hours and degenerated rapidly, whereas the intermediate segment of the dendrites swelled moderately and became more electron lucent. No degenerative changes were evident in pyramidal cell axons and boutons until one day after kainic acid treatment. Less than one hour after kainic acid administration, astrocytes in the CA3 area swelled, initially in the vicinity of the cell body and mossy fiber layers. It is suggested that the paroxysmal discharges initiated in CA3 pyramidal cells by kainic acid served as the stimulus for this response. Phagocytosis commenced between one and three days after kainic acid administration, but remained incomplete at survival times of 6-8 weeks. Astrocytes, microglia, and probably oligodendroglia phagocytized the degenerating material. These results point to the pyramidal cell body and possibly also the dendritic spines as primary targets of kainic acid neurotoxicity. In conjunction with other data, they support the view that lesions made by intraventricular kainic acid can serve as models of epileptic brain damage.

Animals↗

The activation of the renin--angiotensin system in the dog after injection of noradrenaline into the lateral brain ventricle.

Experiments were made on 15 mongrel dogs of either sex. Biologically active substances circulating in the blood after administration of noradrenaline (NA) into the left lateral brain ventricle of the dog were detected using the blood bathed organ technique of Vane. Plasma renin activity was also estimated. NA injected intraventricularly (ivc) in doses of 50-500 mug activated the renin--angiotensin system. Phentolamine prolonged the effect of NA. Propranolol abolished this central effect of NA. The increase in the concentration of endogenous NA in the brain by desmethylimipramine and RO 4-1284 also caused an activation of renin--angiotensin system.

Angiotensin II↗

Hyperglycaemia: imitating Claude Bernard's piqûre with drugs.

Hyperglycaemia lasting for hours, has been produced in unanesthetized cats, rabbits and rats by injection into the cerebral ventricles or the cisterna magna of a variety of drugs (morphine, etorphine, pethidine, beta-endorphin, enkephalin, bombesin, TRH, cholecystokinin, naloxone, propranolol, phentolamine, chloralose, magnesium chloride and GABA). These drugs probably act at the ventral surface of the brainstem and initiate a sympathetic discharge to the adrenals which results in a prolonged release of relatively small amounts of adrenaline. When adrenaline is released in this way hyperglycaemia may be the only effect. The mechanism of the piqûre hyperglycaemia of Claude Bernard may be the same, although Bernard assumed that it resulted from an effect on the floor of the fourth ventricle, i.e. on the dorsal surface of the brainstem. However, it is clear from his description that his trochar not only pricked the floor of the fourth ventricle but penetrated to the ventral surface of the brainstem. Release of adrenaline from the adrenals is usually regarded as a stress response, as in fight, flight, fear or rage when it is suddenly released in large amounts and produces its typical cardiovascular and ocular reactions. The results now obtained with drugs injected intraventricularly or intracisternally suggest an additional physiological role for adrenaline when it is released over prolonged periods and in relatively small amounts producing only hyperglycaemia. Such a release may play a role in the day-to-day control of blood glucose, and its disturbance might underlie non-insulin-dependent diabetes.

Adrenergic beta-Antagonists↗

The role of the lymphatic system in drainage of cerebrospinal fluid and aqueous humour.

1. The jugular lymphatic trunks were cannulated in anaesthetized rabbits and cats. Over 6-8 hr, the mean lymph flow was 2.3 microliters min-1 in the rabbit (one side only) and 5.0 microliters min-1 in the cat (sum of both sides). 2. After a single injection of radio-iodinated albumin into a lateral cerebral ventricle without significant rise in pressure, a mean of 14.4% of the radioactivity was recovered in deep cervical lymph of one side in the rabbit and of 12.9% in that of both sides in the cat. 3. During slow infusion of [125I]albumin and fluorescent dextran of 150,000 mol. wt. into a lateral ventricle of the cat at 20 microliters min-1, radioactivity and fluorescence reached plateaus in deep cervical lymph at 47.4 and 50.0% of their concentrations in cisternal c.s.f. respectively. 4. No significant radioactivity, other than from blood, was detected in superficial cervical lymph after intraventricular injection of radio-iodinated albumin in the cat. 5. No significant radioactivity, other than from blood, was detected in deep cervical lymph of the rabbit or in deep and superficial cervical lymph of the cat within 6 hr after injection of radio-iodinated albumin into the aqueous humour or orbital fat. 6. Gradients of radioactivity in tissues within the orbit suggested that there is a small flow of c.s.f., 0.05-0.15 microliters min-1 in the rabbit, passing centrifugally along the subarachnoid space of the optic nerve, through the posterior part of the globe and into the orbital tissue. Also a small proportion of aqueous humour, 1-2% or more, drains through the anterior sclera into the surrounding tissue. 7. A substantial quantity of cerebrospinal fluid drains into the deep cervical lymphatic system of the rabbit, 30% or more, and of the cat, 10-15% or more. The small component of aqueous humour drainage passing through the wall of the glove does not enter cervical lymph within 6 hr, if at all.

Animals↗

Centrally-induced vasopressor responses to ouabain are augmented in spontaneously hypertensive rats.

Dihydroxyouabain (ouabain), 1.0-10 micrograms, per rat, injected intracerebroventricularly, produced dose-related vasopressor responses accompanied by corresponding increases in abdominal sympathetic nerve activity in 16 weeks old Wistar (NT) rats anesthetized with urethane. The heart rate then also increased, dose-dependently, to ouabain injected in doses up to 10 micrograms. However, 100 micrograms produced arrhythmia resulting in bradycardia. Pressor effects were appreciable within one minute after the ouabain injection, but did not become maximal until between 7-10 min later. Either the removal of sympathetic vasomotor tone by surgical section of the spinal cord or intravenous pretreatment with a vasopressin antagonist significantly reduced the vasopressor responses in the NT rat. Ouabain, 10 micrograms, injected intraventricularly in 16 weeks old Kyoto Wistar rats produced similar cardiovascular responses to those in the NT rat, but the magnitude of the blood pressure responses, along with the heart rate and sympathetic responses, was larger in SHR than in WKY. These results suggest that dihydroxyouabain acts centrally to elevate the blood pressure by increasing not only the sympathetic discharge but also, perhaps, the secretion of vasopressin. In light of previous studies showing that SHRs exhibit both sympathetic hyperactivity and hypersecretion of vasopressin, the present results suggest that their enhanced responsiveness to ouabain could result from both the sympathetic hyperactivity and an enhanced vasopressin release as a result of the centrally injected ouabain.

Animals↗

Characteristics of bombesin-induced inhibition of intake of ethanol.

The temporal and neural dependencies of the inhibitory effect of the administration of bombesin tetradecapeptide (BBS) on the intake of ethanol were assessed in the water-deprived rat. Variation of the intraperitoneal (i.p.) injection of neuropeptide--5% ethanol access interval (0-20 min), revealed that suppression induced by bombesin (0.5-4.0 micrograms/kg) was significantly greater and more potent at shorter intervals. The intake of ethanol was less in rats with subdiaphragmatic vagotomies, but bombesin equivalently suppressed the intake. Intracerebroventricular injection of bombesin more potently and completely inhibited the intake of ethanol but bombesin injected intraventricularly, unlike that given intraperitoneally, elicited excessive grooming and scratching behavior. The suppressant effect of bombesin, given intraperitoneally, requires close temporal contiguity of administration and caloric solution access, which is consistent with a satiety action of a neuropeptide. This satiation effect to ethanol of peripherally administered bombesin appears to reflect a non-vagal, extra-ventricular neural action.

Alcohol Drinking↗