Phenobarbital and 6-aminonicotinamide effect on cerebral enzymatic activities related to energy metabolism in different rat brain areas

The effect of phenobarbital (100 mg/kg i.p.) and 6-aminonicotinamide (6AN) (35 mg/kg i.p.) on enzyme activities related to energy transduction was investigated on the homogenate in toto, non-synaptic mitochondrial fraction and synaptosomal fraction isolated from different rat brain areas (cerebral cortex, hippocampus, hypothalamus, striatum, and medulla oblongata). 6AN treatment decreased: (a) phosphofructokinase in all the areas tested; (b) lactate dehydrogenase on the homogenate in toto in striatum and hypothalamus, and on the synaptosomal fraction in cerebral cortex and corpus striatum; (c) succinate dehydrogenase on non-synaptic mitochondrial fraction in hippocampus and striatum. Finally, aspartate aminotransferase was increased on non-synaptic mitochondrial fraction in striatum and medulla oblongata. Phenobarbital treatment induced an increase of total NADH cytochrome c reductase on mitochondrial fraction in hippocampus and hypothalamus, and a decrease of cytochrome oxidase activity on non-synaptic mitochondrial fraction in hypothalamus and medulla oblongata.     

Regional and subcellular distribution of taurine-synthesizing enzymes in the rat central nervous system

The distribution of cysteine oxidase (CO) and cysteine sulfinate decarboxylase (CSD) was examined in 12 regions of the rat central nervous system (CNS). The distribution of CO activity, expressed as mol of cysteine sulfinate formed per h per g, was the following: hypothalamus, superior and inferior colliculi, 94–99 mol/h/g; olfactory bulbs, cerebral cortex, striatum, and hippocampus, 44–51 mol/h/g; cerebellum, 71 mol/h/g; pons-medula and spinal cord, 94 and 60 mol/h/g, respectively. The distribution of CSD activity expressed as mol of cysteine sulfinate decarboxylated per h per g was the following: hypothalamus and colliculi, 14–21 mol/h/g; olfactory bulbs, cerebral cortex, striatum, hippocampus, and cerebellum, 8–13 mol/h/g; pons-medulla, 7.3; and spinal cord, 3.6 mol/h/g. No CSD activity was detected in sciatic nerve. The subcellular distribution of CO and CSD activities was studied in hypothalamus, colliculi, and cerebral cortex. CO activity was localized in synaptosomes, mitochondria, and microsomes. CSD was primarily confined to the crude mitochondrial fraction and after subfraction, recovered mainly in the synaptosomal fraction.     

Effects of Sex Hormones,Forskolin, and Nicotine on Choline Acetyltransferase Activity in Human Isolated Placenta

The activity of choline acetyltransferase (ChAT) was investigated in the human placenta before and after long-term incubation (24 h) to test the effects of sex hormones, nicotine and forskolin. ChAT activity differed considerably between the amnion (0.03 mol/mg protein/h) and the villus (0.56). After long-term incubation, ChAT activity persisted in the latter but declined in the amnion. Neither sex hormones (-estradiol, testosterone, progesterone; 10 or 100 nM each) nor follicle stimulating hormone and luteinizing hormone (FSH/LH; 8.4 U/ml each) modified ChAT activity. Also nicotine (1 nM–100 M) did not affect ChAT activity. Forskolin, an activitor of adenylyl cyclase, reduced ChAT activity in the villus but not in amnion. The present model offers the possibility to investigate ChAT regulation in intact tissue under long-term incubation. The risks of maternal smoking during pregnancy cannot be attributed to an effect of nicotine on placental ChAT activity. Differences in the regulation of ChAT appear to exist between neuronal and nonneuronal cells.     

Regional distribution of membrane-bound γ-glutamyl transpeptidase activity in mouse brain

Activity of membrane-bound -glutamyl transpeptidase (-GTP) was examined in various regions of mouse brain, in capillaries of the cerebral cortex and in telencephalic choroid plexuses. The level of activity in the capillaries was double and that of the choroid plexus nine times that of the -GTP activity found in the brain, septum, hippocampus, hypothalamus, thalamus, cerebellum, frontal cortex, pons, medulla oblongata, and amygdala. Histochemically the -GTP activity was demonstrated in the surface membranes of choroidal cells and in the endothelium of small capillaries.The activities of -GTP of cerebral cortex, choroid plexus, and capillaries from rabbit were 5–17 times greater than those from corresponding areas of mouse brain. While 30 mM methionine stimulated (in vitro) the enzyme from mouse brain, no such effect was observed with the enzyme activity from rabbit brain. The -GTP activity from the capillaries of cerebral cortex of both mouse and rabbit was not effected by the presence of methionine.These findings suggest existence of differences in the specificity of -GTP activity in these two species.     

Considerations on the astroglial architecture and the columnar organization of the cerebral cortex

Present knowledge on astroglial roles in brain organization and function indicates that these cells can regulate the extracellular ionic composition and modulate neuronal activity. In this regard, the panglial synctium formed by electrically and chemically coupled stellate astrocytes (general mammalian architecture) is believed to provide an intracellular pathway for the redistribution of ions and molecules within the cerebral cortex. Long astroglial interlaminar processes (primate-specific architecture) that run parallel to apical dendrites in the cerebral cortex of primates, may provide additional properties to the glial participation in cortical physiology and function. Since these processes are exclusively present within the primate order, functional models of cortical computations for these species should incorporate the astroglial interlaminar architecture in addition to the panglial synctium. This study analyzes possible implications of interlaminar astroglial processes, for the regulation of the extracellular ionic composition and segregation of functional columns in the cerebral cortex.     

Insulin-like immunoreactivity in the human brain

Summary The localization and regional distribution of insulin-like immunoreactivity (IRI) was studied in human brain autopsy material using the indirect immunofluorescence technique. A positive reaction for IRI could be observed in many neurons of the hypothalamus, the hippocampus, corpus amygdaloideum, medulla oblongata (especially within the nuclei of cranial nerves IX, X and XII), and the cerebral cortex, whereas the cerebellar cortex was lacking in immunohistochemically detectable insulin-like material. No nerve fibres containing polypeptides could be revealed. Additionally, the inuslin content of various brain regions was estimated by radioimmunosassay. Insulin concentrations in human nervous tissue were found to be elevated in comparison to blood plasma levels.The present investigation was supported by the HFR Neurobiologie und Hirnforschung and the HFR Diabetes mellitus und Fettstoffwechselstörungen of the GDR (Ministries of Higher Education and Health respectively) and the Finnish Ministry of Education     

A comparison of male and female recombination frequency in wheat using RFLP maps of homoeologous group 6 and 7 chromosomes

A novel approach was used to compare male and female recombination rates in wheat. Doubled haploid lines were developed from an F₁ using two distinct approaches: the anther-culture technique and the Hordeum bulbosum system, from which sets of lines were developed from male and female meioses, respectively. The genotype of the lines was established at RFLP and isozyme markers polymorphic on chromosomes of homoeologous groups 6 and 7, and male and female linkage maps were calculated using this information. The markers in one segment of chromosome 6B exhibited disturbed segregation frequencies in the anther-culture population. The male and female maps differed significantly in recombination frequency between some markers on two chromosomes, and these were consistent in direction within chromosomes and inconsistent in direction between chromosomes. In two of the four chromosomes studied the male map was much longer than the female map. These results suggest that significant differences may exist in male and female recombination frequencies in bread wheat which are specific to certain chromosomal segments but are inconsistent in direction between chromosomes. Other factors, such as environmental influences, may also be important in creating differences.     

Changes of mitochondrial cytochrome c oxidase and FoF1 ATP synthase subunits in rat cerebral cortex during aging

The contents of subunits I, II/III, and IV of cytochrome c oxidase and of subunits , and of FoF1 ATP synthase in inner mitochondrial membrane proteins purified from cerebral cortex of rat at 2, 6, 12, 18, 24, and 26 months of age were analyzed by western blot. Age-related changes in the content of subunits, either of mitochondrial or nuclear origin, were observed. All the cytochrome c oxidase (COX) subunits examined showed an age-related increase from 2-month-old rats up to 24 months with a decrease at the oldest age (26 months). The same pattern of age-dependent changes was observed for ATP synthase, while the and subunits increased progressively up to 26 months.     

Protein synthesis rates in rat brain regions and subcellular fractions during aging

In vivo protein synthesis rates in various brain regions (cerebral cortex, cerebellum, hippocampus, hypothalamus, and striatum) of 4-, 12-, and 24-month-old rats were examined after injection of a flooding dose of labeled valine. The incorporation of labeled valine into proteins of mitochondrial, microsomal, and cytosolic fractions from cerebral cortex and cerebellum was also measured. At all ages examined, the incorporation rate was 0.5% per hour in cerebral cortex, cerebellum, hippocampus, and hypothalamus and 0.4% per hour in striatum. Of the subcellular fractions examined, the microsomal proteins were synthesized at the highest rate, followed by cytosolic and mitochondrial proteins. The results obtained indicate that the average synthesis rate of proteins in the various brain regions and subcellular fractions examined is fairly constant and is not significantly altered in the 4 to 24-month period of life of rats.A preliminary report of these results was previously presented at: WFN-ESN Joint Meeting on: Cerebral Metabolism in Aging and Neurological Disorders, Baden, August 28–31, 1986.     

Complex sex-determining mechanisms in three species of South American grasshoppers (Orthoptera,Acridoidea)

The karyotype of three species of South American grasshoppers are studied in this paper. Leiotettix sanguineus has two chromosome races, one of them with 2n=23 and an XO sex mechanism and the other, as far as we know limited to the Cerro Chato population, with 2n=22 and an XY sex mechanism. Leiotettix politus has two kinds of individuals, one with 2n=14 and XY sex chromosomes and the other 2n=13 and an X₁X₂Y mechanism. Dichroplus dubius presents 2n=21 and an X₁X₂Y sex chromosomes. One of the three specimens studied shows aberrant behaviour in the meiotic process.     

The effects of adrenalectomy and thermal stress on glutamic acid decarboxylase activity in different regions of the rat brain

Glutamic acid decarboxylase (GAD) enzyme activity was measured in synaptosomes prepared from the hypothalamus, the hippocampus, the striatum and the cerebral cortex of control, adrenalectomized and rat exposed to a thermal stress. Adrenalectomy caused a statistically significant decrease in the enzyme activity in the striatum, while it had no effect in the other three brain areas. On the other hand, exposure to the thermal stress resulted in a dramatic increase of GAD specific activity in all brain areas examined. This thermal stress-induced increase in enzyme activity was observed in both non-operated and adrenalectomized animals, which implies that it is not mediated by glucocorticoids.Abbreviations used GAD glutamic acid decarboxylase - GABA -aminobutyric acid - AET 2-aminoethylisourethonium bromide - ADX adrenalectomized - rpm revolutions per minute     

Glutamic acid decarboxylase-positive neuronal cell bodies and terminals in the human cerebellar cortex

The distribution of -aminobutyric acid (GABA) in the human cerebellar cortex was studied using immunohistochemistry for glutamic acid decarboxylase (GAD), the enzyme that catalyses GABA synthesis. Observations by light microscopy revealed, in all layers of the cerebellar cortex, strong, punctate positivity for GAD, related to putative GABAergic nerve terminals, as well as a diffuse cytoplasmic immunoreactivity within neuronal cell bodies. GAD-positive nerve terminals were found in close relationship with the walls of the cerebellar cortex microvessels. Observations by electron microscopy revealed positive nerve terminals in contact with the astrocyte perivascular sheath of capillaries. GAD immunoreactivity was also detected within astroglial perivascular endfeet and endothelial cells. The findings provide further insights into the GABAergic synapses of the circuitry of the human cerebellar cortex. The detection of vascular GAD immunoreactivities suggests that GABAergic mechanisms may regulate cerebellar microvessel function.     

Insulin immunohistochemistry of rodent CNS: Apparent species differences but good correlation with radioimmunological data

Summary Using an indirect immunofluorescence technique the distribution of insulin was mapped in brains of Wistar strain rats and mice. Insulin immunoreactivity was found to be widely distributed throughout mouse CNS, whereas in rat brain a restriction of immunoreactive material to cerebral blood vessels and ependymal cells and/or tanycytes of the brain ventricles was observed. In radioimmunological studies the amount of insulin (IRI) was estimated for different brain areas (cerebral cortex, brain stem, cerebellum, hippocampus, thalamus and hypothalamus). In the case of Wistar rats very low levels of IRI were found. On the contrary, the same regions in mouse brain contained considerably greater amounts of IRI. The comparison between histochemical and biochemical data revealed a good correlation. It is concluded that part of the insulin measured by radioimmunoassay is associated with neuronal structures.Research supported by the Ministerium für Hoch- und Fachschulwesen der DDR and Ministerium für Gesundheitswesen der DDR     

5,7-Dihydroxytryptamine: Regional Brain Concentrations Following Intraventricular Administration to Rats

The concentrations of 5,7-dihydroxytryptamine (5,7-DHT) and serotonin (5-HT) were measured in brainstem, hypothalamus and cerebral cortex 0, 2, 6, 12, and 24 hours following the bilateral, lateral ventricular injection of 5,7-DHT (100 g/each ventricle) into adult male rats. At 6 hours, 5,7-DHT levels had decreased 99% from 0 hr values in all brain regions. Thereafter, 5,7-DHT levels continued to decline in cortex, but not in hypothalamus or brainstem; at 24 hr, but not 48 hr, 5,7-DHT peaks were still measurable in each brain region examined. Serotonin levels in all three regions also fell markedly by 2-6 hours after 5,7-DHT administration. At 24 hours, hypothalamus and brainstem 5HT levels had declined >70% and cerebral cortex 50% below control values. The relevance of these findings to the protective action of monoamine reuptake blockers is discussed.     

The temperature-dependent duration of development and parasitism of three cereal aphid parasitoids, Aphidius ervi, A. rhopalosiphi, and Praon volucre

Temperature dependencies were established for the egg-to-mummy and mummy-to-adult phases, for mummy mortality, and for parasitism of Aphidius ervi Haliday, Aphidius rhopalosiphi De Stefani-Perez, and Praon volucre (Haliday) (Hymenoptera, Aphidiidae), three parasitoids of Sitobion avenae (Fabricius) (Homoptera, Aphididae), at 8°C, 12°C, 16°C, 20°C, and 25°C on winter wheat (cv. Haven). A physiological model described temperature-dependent development over the full temperature range, whereas a linear model was fitted for data above 8°C and used to estimate the lower temperature thresholds and day-degrees (° D) required for development. The thresholds for A. ervi were 2.2°C for egg-mummy development and 6.6°C for mummy-adult development, those for A. rhopalosiphi were 4.5°C and 7.2°C, and those for P. volucre were 3.8°C and 5.5°C. The time to develop into mummies and adults differed significantly between the three species: A. ervi development into mummies required an average of 159 ° D, while development into adults took an average of 73 ° D. The corresponding average times required for A. rhopalosiphi and P. volucre to develop mummies were 124° D and 126° D, while their development into adults required an average of 70° D and 150° D, respectively. Mummy mortality was 25–35% at 8°C and less at the higher temperatures tested, but began to increase again at 25°C, showing a quadratic relationship between mortality and temperature. Parasitization was very low or, in the case of P. volucre, absent up to 12°C and thereafter increased with increasing temperature. The relationship between parasitization, recorded as percent aphids mummified, and temperature was linear at the temperatures tested and depended on species. A. ervisuperparasitized 11.1% aphids at 20°C and 16.6% aphids at 25°C, whereas superparasitism was low in A. rhopalosiphi and absent in P. volucre. From 16°C to 25°C the P. volucre sex ratio increased. For A. ervi and A. rhopalosiphi there was no trend with temperature, but at 20°C and 25°C it was close to even. Field data for 1996 and 1997 allowed for a comparison of actual and expected emergence of overwintering mummies. In both years, parasitoids were predicted to have emerged from overwintering mummies well in advance of the onset of aphid infestation, and more than a month earlier than the first parasitized aphids were found in winter wheat. Observations from trap plants in other crops supported the predictions of the models. Other factors that can affect biological control by cereal aphid parasitoids are discussed.     

The selective inhibition of hippocampal glutamic acid decarboxylase in zinc-induced epileptic seizures

The intracerebroventricular administration of Zn²⁺ (0.3 mol/10 l) causes epileptic seizures characterized by running fits, jumping, vocalization, fasiculation of facial muscles, myoclonic movements of the limbs and tonic-clonic convulsions. These episodes are blocked or reversed by -aminobutyric acid (0.4 mol/10 l). When assayed under conditions where pyridoxal phosphate was not added, the activity of glutamic acid decarboxylase decreased significantly in hippocampus from 18.9 to 15.3 and 9.7 mol¹⁴CO₂ formed/gram proteins/20 min, 15 and 30 min following administration of Zn²⁺. The inhibition of glutamic acid decarboxylase by Zn²⁺ was selective occurring only in hippocampus and not in the hypothalamus, amygdala, caudate or thalamus. The inhibition of glutamic acid decarboxylase was not due to a reduction in the concentration of endogenous pyridoxal phosphate which remained unaltered in hippocampus following Zn²⁺ administration.     

Polyamines and the development of isolated neurons in cell culture

The possible role of polyamines in the development of isolated neuroblasts from the cerebral cortex of embryonic chick brain was studied by means of three enzyme activated irreversible inhibitors of ornithine decarboxylase. -Difluoromethylornithine (MDL 71782) showed no effects on development at doses which depleted dramatically neuronal putrescine and spermidine levels. In contrast, the two other inhibitors, (E)--(fluoromethy)dehydroputrescine (MDL 72197) and 6-heptyne-2,5-diamine (MDL 72175) blocked the formation of neuronal outgrowths completely at 100 M and higher concentrations. Their effects on neuronal polyamines differed at this concentration considerably. The growth inhibitory effect of the ornithine decarboxylase inhibitors was in all cases reversible: cells which were grown after 3 days of exposure to the drugs in normal medium produced neuronal networks. The presence of putrescine at 10M concentration in the culture medium prevented the growth inhibitory effect of 100 M concentrations of the drugs. This concentration of putrescine was not only capable of preventing, but also of reversing growth inhibition by the ornithine decarboxylase inhibitors. Although the cellular polyamine levels were not correlated with the morphological development of chick embryo cortical neurons, the present study leaves no doubt that putrescine plays an essential role in neuronal differentiation.     

The effect of partial noradrenergic denervation on corticosterone secretion in the rat

DSP₄ (N-(-2-chloroethyl)-N-ethyl-2-bromobenzylamine) treatment significantly decreased the noradrenaline content in the hippocampus, frontal cortex and hypothalamus of the rat brain. DSP₄ treatment did not affect plasma corticosterone levels. Clonidine, an ₂-adrenoceptor agonist, had no effect on corticosterone secretion in either DSP₄- or saline-treated rats. Isoproterenol, a -adrenoceptor agonist, significantly stimulated corticosterone secretion. This effect was inhibited by the prior administration of the -adrenoceptor antagonist propranalol. DSP₄ treatment did not alter the isoproterenol-induced stimulation of corticosterone secretion. The administration of a high dose of dexamethasone (100 g/kg, i.p.) significantly decreased the plasma corticosterone concentration of saline-treated controls, while an intermediate dose (25 g/kg, i.p.) did not suppress corticosterone release significantly. DSP₄-treatment did not influence dexamethasone-induced suppression of corticosterone secretion. These results show that significant decreases in noradrenaline content in the hippocampus, frontal cortex and hypothalamus appear to have no effect on the regulation of corticosterone secretion and that corticosterone secretion may be stimulated by catecholamines via -adrenoceptors.     

Developmental and Regional Alteration of κ-Opioid Receptors in Seizure-Susceptible EL Mouse Brain

The binding of [³H]ethylketocyclazocine ([³H]EKC) under the suppression of and sites in the brain of EL mice (seizure-susceptible) was examined to determine the relationship between seizures and the dynorphinergic system. The density of -opioid receptors in the cerebrum of adult EL mice during interictal periods significantly increased, without changes in apparent affinities, compared with that of adult ddY mice (seizure-nonsusceptible; the mother strain of EL). Subsequently, the binding of 0.8 nM [³H]EKC in 8 brain regions was examined in the 2 strains. The [³H]EKC binding in 25-day-old EL mice that had no seizures significantly increased in the hippocampus and amygdala. At the age of 50 days, EL mice displayed abortive seizures, and the number of sites in EL mice was significantly greater in the hippocampus, amygdala and cerebral cortex. It was further shown that the binding of [³H]EKC in 150-day-old adult EL mice during interictal periods was markedly increased in the hippocampus, amygdala, cerebral cortex and striatum, compared with the corresponding regions in ddY mice. The up-regulation of receptors in the EL mouse brain may result from deficits in endogenous dynorphins, which could be involved in the pathogenesis of seizure diathesis and seizure manifestations in the EL mouse.     

Hypothalamus and cytodifferentiation of the foetal pituitary gland

Summary To investigate whether the hypothalamus is involved in the cytodifferentiation of the anterior pituitary gland, rat foetuses were encephalectomized in utero on day 16 of pregnancy.Pituitary sections from encephalectomized and normal littermate foetuses were studied on day 21 with the immunofluorescence technique using antibodies and -MSH, anti -MSH, anti -(17–39) ACTH and anti -(1–24) ACTH. On day 16, only the anti -MSH revealed a few cells in the pars distalis but not in the pars intermedia. On the other hand, on day 21, the pituitary cells reacting with antibodies anti -MSH, anti -MSH and anti -(17–39) ACTH were as numerous in the encephalectomized foetuses as in the normal littermate foetuses. The cells revealed with the antibody anti -(1–24) ACTH were less numerous and less fluorescent in the pars distalis and intermedia of the hypophysis of the encephalectomized foetuses.On day 21, the adrenals of the encephalectomized foetuses were atrophied in comparison with those of the normal littermate foetuses but they were larger than on day 16.These data suggest that the cytodifferentiation of the corticotroph and melanotroph cells of the hypophysis occurs without the influence of the hypothalamus which is necessary for the normal release of ACTH.