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Distribution of neuronal nitric oxide synthase-immunoreactive neurons in the cerebral cortex and hippocampus during postnatal development.

Although many reports have argued a role for nitric oxide (NO) during postnatal development, there has been no combined demonstration in the cerebral cortex and hippocampus. We have investigated the distribution and morphology of neurons and fibers expressing neuronal NO synthase (nNOS) in the cerebral cortex and hippocampal formation of rats during the postnatal development, and correlated these findings with developmental events taking place in these regions. In the cerebral cortex, the nNOS-immunoreactive cells could be divided into two classes : heavily stained neurons and lightly stained neurons. For the lightly stained nNOS-positive neurons, only the cell bodies were observed, whereas for the heavily stained neurons, the cell bodies and their dendrites were visible. During the postnatal days, heavily stained neurons reached their typical morphology in the second week and appeared in all layers except for layer I. In the hippocampus, there was a transient expression of nNOS in the pyramidal cell layer at P3-P7, and this expression disappeared during following days. The adult pattern of staining developed gradually during the postnatal period. This study suggested that these alterations might reflect a region-specific role of NO and a potential developmental role in the postnatal cerebral cortex and hippocampus.

Animals↗

Early asymmetry of gene transcription in embryonic human left and right cerebral cortex.

The human left and right cerebral hemispheres are anatomically and functionally asymmetric. To test whether human cortical asymmetry has a molecular basis, we studied gene expression levels between the left and right embryonic hemispheres using serial analysis of gene expression (SAGE). We identified and verified 27 differentially expressed genes, which suggests that human cortical asymmetry is accompanied by early, marked transcriptional asymmetries. LMO4 is consistently more highly expressed in the right perisylvian human cerebral cortex than in the left and is essential for cortical development in mice, suggesting that human left-right specialization reflects asymmetric cortical development at early stages.

Adaptor Proteins, Signal Transducing↗

Ethanol induces Fas/Apo [apoptosis]-1 mRNA and cell suicide in the developing cerebral cortex.

INTRODUCTION: Animal studies modeling fetal alcohol syndrome have demonstrated that developmental exposure to alcohol is associated with decreased brain weight and significant neuronal loss in multiple regions of the developing brain. Our previous data suggest that the Fas/Apo [apoptosis]-1 receptor is transiently expressed in the developing cerebral cortex during the peak period of naturally occurring apoptotic cell death and maximum sensitivity to alcohol. Therefore, we hypothesized that ethanol increases the expression of suicide receptors such as Fas/Apo-1 in the developing fetal cerebral cortex and leads to an upregulation or extension of the normal period of apoptosis and consequent disorganization of the neural circuitry. METHODS: Ethanol was administered in one of four doses (120, 320, 630, and 950 mg/dl) to organotypic explant cultures of the developing cerebral cortex established from postnatal day 2 rats and maintained for 6 days in vitro. The number of cells expressing Fas/Apo-1 receptor mRNA was counted. Apoptosis was measured by the use of two independent assays; a cell death enzyme-linked immunosorbent assay for DNA fragmentation and flow cytometric analysis of Annexin-V binding to phosphatidylserine externalized to the outer leaflet of the plasma membrane. Necrosis was also estimated by two independent measures, the amount of lactate dehydrogenase released into culture medium and flow cytometric analysis of cells that were positive for both Annexin-V and propidium iodide. RESULTS: A significantly larger number of developing cortical cells expressed Fas/Apo-1 mRNA at the lower doses (120 and 320 mg/dl) than at the higher doses (630 and 950 mg/dl). Furthermore, ethanol induced apoptosis in a dose-related manner, with peak apoptosis observed at a dose of 630 mg/dl in the case of DNA fragmentation and at 630 and 950 mg/dl in the case of phosphatidylserine translocation to the outer leaflet of the plasma membrane. Ethanol did not induce necrosis at any of the administered doses of ethanol. CONCLUSIONS: Our data suggest that ethanol induces a susceptibility to apoptotic signals at low doses by upregulating the expression of mRNAs for cytotoxic receptors such as Fas/Apo-1 in the developing cerebral cortex. However, ethanol itself specifically induces apoptosis in the developing cerebral cortex only at higher doses.

Animals↗

Morphometric study of human cerebral cortex development.

Morphometric studies of immature cerebral cortex in humans show developmental changes extending up to the time of adolescence. Growth of dendrites and of synaptic connections occurs during infancy and early childhood. Excess synaptic connections are eliminated during later childhood years. The exuberant connections that occur during infancy may form the anatomical substrate for neural plasticity and for certain types of early learning.

Adolescent↗

Binding of [3H]-pilocarpine to membranes from rat cerebral cortex.

Binding of [3H]-pilocarpine to synaptic membranes in rat cerebral cortex was investigated, pilocarpine binding was also studied by competition of unlabelled pilocarpine with the [3H]-labelled muscarinic antagonist, [3H]-N-methyl-4-piperidinyl benzilate. 1. [3H]-pilocarpine binding sites are of protein nature, and the highest specific activity of binding is found in the synaptosomal fraction of all subcellular fractions. 2. Competition studies show that only muscarinic drugs inhibit [3H]-pilocarpine binding in their pharmacologically active concentration range. 3. Binding of [3H]-pilocarpine is influenced by GMPP-(NH)P (0.1 mM) similarly to the binding of other muscarinic agonists. 4. Examination of pilocarpine binding with [3H]-pilocarpine and the competition experiments with unlabelled pilocarpine indicate the presence of three sites with different affinities: 5 nM, 0.2 microM and 30 microM respectively. 5. Experiments with [3H]-4-NMPB and [3H]-pilocarpine indicate that there are more [3H]-pilocarpine binding sites than [3H]-4-NMPB binding sites in rat cerebral cortex.

Animals↗

Characterization of beta-adrenergic receptors in duck cerebral cortex.

beta-adrenoceptor binding sites were characterized in duck cerebral cortex by an in vitro binding technique, using [3H]dihydroalprenolol ([3H]DHA) as a receptor-specific radioligand. The specific binding of [3H]DHA to duck cerebral cortical membranes was found to be rapid, stable, saturable, reversible, and of high affinity. Saturation analysis resulted in a linear Scatchard plot suggesting binding to a single class of receptor binding sites with high affinity (Kd = 1.18 nM) and high capacity (Bmax = 162 fmol/mg protein). Competition studies showed the following relative rank order of potency of various compounds to inhibit the [3H]DHA binding: antagonists--ICI 118,551 > S(-)-propranolol >> betaxolol, yohimbine, WB-4101, prazosin, mianserine; agonists--isoprenaline approximately equal to fenoterol > salbutamol >> clonidine, phenylephrine. The obtained data suggest that in duck cerebral cortex beta-adrenergic receptors (like those described in brains of chick and pigeon) are of the beta 2 subtype. This is in contrast to what has been reported for the mammalian brain, where--among beta-adrenoceptors--the beta 1 subtype is predominant.

Adrenergic beta-Antagonists↗

Regionalization of the cerebral cortex: developmental mechanisms and models.

The cerebral cortex is composed of functionally specialized areas that have unique connections with other cortical targets and subcortical nuclei. The developmental mechanisms responsible for the formation of discrete regions must include the regulation of the expression of genes encoding proteins that control axon guidance and targeting. New data on patterns of gene expression demonstrate the early appearance of such guidance molecules, thus reflecting the early emergence of regional specification within the cortex. Transplant and cell culture studies suggest that the decisions made by neuronal progenitor cells to express region-appropriate phenotypes is controlled by the capacity of the cells to respond to and have access to specific signals. The key to understanding cortical specification may lie in determining the factors that control receptor diversity on progenitors and the temporal and spatial distribution of inductive signals within the forebrain.

Animals↗

[Morphometric studies on the effect of 5-hydroxytryptamine creatinine sulfate and noradrenaline bitartrate on the growth of cerebral cortex explants in vitro].

Explants of cerebral cortex of neonatal rats were cultivated up to 15 DIV being treated with 5-hydroxytryptamine and noradrenaline, resp. Effects of exogenous transmitters on the growth of explants were investigated by means of morphometrical methods. Different effects on the growth depending on the duration of cultivation and the type of transmitter administered were found. Stimulatory influences have been seen to occur up to 4 DIV (noradrenaline) and up to 2 DIV (serotonin).

Animals↗

gamma-Aminobutyric acid inhibition of histamine-induced inositol phosphate formation in guinea-pig cerebellum: comparison with guinea-pig and rat cerebral cortex.

1. gamma-Aminobutyric acid (GABA), 2 mM, inhibited basal accumulation of [3H]-inositol monophosphate ([3H]-IP1) in lithium-treated slices of guinea-pig cerebellum preincubated with [3H]-inositol. In contrast, 2 mM GABA stimulated the accumulation of [3H]-IP1 in rat cerebral cortical slices over a 60 min incubation period, but had no significant effect in slices of guinea-pig cerebral cortex. The estimated IC50 for the inhibitory action of GABA in guinea-pig cerebellar slices was 0.52 +/- 0.12 mM. 2. GABA inhibited histamine-induced [3H]-IP1 accumulation in guinea-pig cerebellar slices in a non-competitive manner. The best-fit value for the maximum level of inhibition was 74 +/- 6%. The estimated IC50 for GABA was 0.77 +/- 0.15 mM and was not significantly different from the IC50 for inhibition of the basal accumulation of [3H]-IP1. The response to histamine in guinea-pig and rat cerebral cortical slices was also inhibited by 2 mM GABA. 3. In guinea-pig cerebellar slices 2 mM GABA potentiated histamine-induced [3H]-inositol bisphosphate ([3H]-IP2) accumulation, whereas in both guinea-pig and rat cerebral cortex the effect was inhibition. 4. Isoguvacine and muscimol, GABAA-selective agonists, and (-)-baclofen, GABA(B)-selective, had no significant effect on basal or histamine-stimulated accumulation of [3H]-IPs in guinea-pig cerebellar slices. (-)-Baclofen had only a weak inhibitory effect on [3H]-IP1 accumulation in guinea-pig-cerebral cortex (16 +/- 6% inhibition with 10 microM (-)-baclofen), whereas in rat cerebral cortex (-)-baclofen mimicked the inhibitory effect of GABA. 5. Nipecotic acid (1 mM) had qualitatively similar effects to those of 2mm GABA in guinea-pig cerebellar slices. 6. The competitive GABA uptake inhibitors SK&F 89976-A, SK&F 100330-A and SK&F 100561-A were potent histamine H,-receptor antagonists, as indicated by the inhibition of [3H]-mepyramine binding to homogenates of guinea-pig cerebellum and cerebral cortex. 7. GABA (2 mM) caused a small inhibition (12 + 3%) of [3H]-inositol incorporation into total inositol phospholipids in guinea-pig cerebellar slices, as in rat cerebral cortical slices, whereas 0.2mm histamine caused a small stimulation (15 + 4%). In the presence of both GABA and histamine, [3H]-inositol incorporation was unchanged from basal (101 + 5%). 8. GABA also inhibited [3H]-IP1 formation induced by endothelin-1 in guinea-pig cerebellar slices and increased, but not significantly, the amount of [3H]-IP2 accumulated. This, taken with the inhibitory effect on basal and histamine-stimulated accumulation, suggests that the action of GABA in guinea-pig cerebellar slices may be non-selective and may not be exerted through a specific GABA receptor.

Animals↗

Cellular physiology of the neonatal rat cerebral cortex.

The early development of the cerebral cortex is characterized by neurogenesis, neuronal migration, cellular differentiation and programmed cell death. Cajal-Retzius cells, developing cortical plate neurons and subplate cells form a transient synaptic circuit which may serve as a template for the formation of cortical layers and columns. These three neuronal cell types show distinct electrophysiological properties and synaptic inputs. Endogenous or exogenous harmful disturbances during this developmental period may lead to the preservation of early cortical circuits, which may act as trigger zones for the initiation of pathophysiological activity.

Action Potentials↗

Ageing affects cytosolic Ca2+ binding proteins and synaptic markers in the retina but not in cerebral cortex neurons of the rat.

Two cerebral cortex areas (frontal and occipital) and the retina of rats varying in age from 0.4 to 30 months were investigated for the expression levels and distribution of two cytosolic high affinity Ca2+ binding proteins, calbindin-28 and calretinin, and of two presynaptic protein markers. Of these latter proteins, one is integral (synaptophysin) the other peripheral (synapsin I) to the synaptic vesicle membranes. In the cortex areas, no significant changes of the markers were observed, except for a drop of calretinin from 0.4 to 2 months, probably related to a stage of neuronal development. In the retina, calbindin-28 decreased progressively during ageing (-40% at 30 months) while calretinin remained unchanged. Concomitantly, the two synaptic vesicle proteins dropped, synaptophysin > 50% and synapsin I > 85%. The role of these changes in sustaining the functional alterations previously described in the retina of aged animals remains to be investigated.

Aging↗

Neuronal (Na+,K+)-ATPase and the release of purines from mouse and rat cerebral cortex.

Slices of mouse or rat cerebral cortex were incubated with [3H]adenine or [3H]adenosine, and [14C]GABA. Purines and GABA could subsequently be released by ouabain. The release of purines previously shown to occur on restoring elevated K+ levels to normal was not mimicked by noradrenaline at concentrations which activate (Na+,K+)-ATPase. Potassium-free solutions evoked no release of purine during the test period, but resulted in a large release when K+ was restored to normal. K+-free solutions evoked an immediate release of GABA. It is concluded that (Na+,K+)-ATPase is not involved in purine release.

Animals↗

Handedness is associated with asymmetries in gyrification of the cerebral cortex of chimpanzees.

Gyrification of the cerebral cortex reflects complexity in cortical folding during development of the brain. In this paper, we evaluated whether chimpanzees show asymmetries in gyrification and if variation in gyrification asymmetries were associated with handedness. Magnetic resonance images were obtained in a sample of 76 chimpanzees, and gyrification measures were obtained from 10 equally spaced slices of the cortex. Asymmetry quotients (AQs) in gyrification were compared for 4 measures of handedness including reaching, coordinated bimanual actions, manual gestures, and throwing. Overall, the chimpanzees showed significant differences between the right and left hemispheres that were region specific. Significant differences in AQ's were found in right- and nonright-handed chimpanzees for throwing and, to a lesser degree, for manual gestures. Increasing age was associated with increasing gyrification in the prefrontal regions, particularly in female chimpanzees. The results indicate that variation in gyrification between hemispheres is associated with functional measures of laterality in chimpanzees.

Aging↗

Androgen binding and metabolism in the cerebral cortex of the developing rhesus monkey.

The influence of gonadal steroids on the maturation of the cerebral cortex and their possible influence on cortical function are not well understood. The present study examines androgen binding and metabolism in regions of the cerebral cortex and several subcortical structures at different time points during the development of the rhesus monkey. Androgen binding and metabolism were assessed in selected cortical and subcortical regions from neonatal, juvenile, and adult rhesus monkeys. Specific high affinity androgen-binding sites (apparent equilibrium dissociation constants for [3H]R1881, 0.04-0.3 nM) were observed in many areas of the rhesus monkey brain. These areas included (but were not limited to) the hypothalamus, amygdala, dorsolateral prefrontal cortex, orbital prefrontal cortex, visual and somatosensory cortex, and corpus callosum. Although the highest level of androgen binding was observed in the hypothalamus (8-20 fmol/mg protein), cortical samples also had measurable levels of binding (1-5 fmol/mg protein). No apparent regional or developmental differences in the number or affinity of androgen-binding sites were detected in animals ranging from 1 week to 8 yr in age. Androgen metabolism via aromatization and 5 alpha-reduction was observed in all regions of the neocortex examined, although at lower levels than in the hypothalamus and amygdala. Overall production of estrogens and 5 alpha-reduced androgens declined approximately 10-fold from prenatal to early postnatal life in both cortical and subcortical structures. The presence of androgen binding and metabolism in the monkey cerebral cortex indicates that steroid hormones may have considerable impact on cortical function in primates at postnatal as well as prenatal ages.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Studies on the transport of carnitine in the brain using synaptosomes isolated from guinea-pig cerebral cortex.

Synaptosomes isolated from guinea pig cerebral cortex accumulate L-carnitine from the medium in an active process, dependent on the sodium gradient across the plasma membrane and on (Na+ + K+)-ATPase activity. L-Carnitine uptake is inhibited by oxidative phosphorylation uncouplers and by ouabain, a known inhibitor of (Na+ + K+)-ATPase. In addition, the omission of Na+ or its replacement by Li+ inhibited the transport, which was also competitively inhibited by gamma-aminobutyrate. The kinetics of carnitine uptake show that the overall process would consist of two components: a passive diffusion and a carrier-mediated transport which is saturated at 1-2 mM carnitine concentration.

Animals↗

Serotonergic axons form basket-like terminals in cerebral cortex.

Serotonergic axons in the posterior cerebral cortex of the cat were demonstrated immunohistochemically using a monoclonal antibody to serotonin (5-HT). This technique reveals the presence of a dense serotonergic innervation of single cortical neurons at the light microscopic level. 5-HT axons with large varicosities (1-6 microns in diameter) form distinct, basket-like arrays around counterstained somata principally in layer I. In each basket one or more axons encircle and make repeated contact with the soma. Some axons extend from the soma and apparently climb along the dendrites of the target neuron. The climbing 5-HT axons form a stellate or horizontal pattern suggesting that the target cells are non-pyramidal neurons of the supragranular layers.

Animals↗

Identification of a ubiquitin- and ATP-dependent protein degradation pathway in rat cerebral cortex.

To investigate the existence of a ubiquitin-dependent protein degradation system in the brain, the proteolytic activity of the cerebral cortex was examined. The soluble extract of rat cerebral cortex degraded 125I-radiolabeled lysozyme in an ATP-dependent manner. The ATP-dependent proteolysis was suppressed with iodoacetamide, which inhibits ubiquitin conjugation, and was abolished by blocking of the amino residues of lysozyme. These results suggest the participation of ubiquitination in the proteolytic activity. An ATP-dependent 125I-ubiquitin-conjugating activity was detected in fraction II from the cerebral cortex. The presence of ATP-dependent proteolytic activity which acted preferentially on ubiquitinated lysozyme was demonstrated, using ubiquitin-125I-lysozyme conjugates as a substrate. The proteinase had a molecular mass of 1500 kDa and displayed nucleotide dependence and sensitivity to various proteinase inhibitors similar to those of the 26S proteinase complex found in reticulocytes. Dialysis of the soluble fraction caused a decrease in the proteolytic activity of ATP-dependent and preferential for ubiquitin-lysozyme conjugates and a reciprocal increase in the ATP-independent free 125I-lysozyme-degrading activity which was scarcely detected before dialysis. The former ATP-dependent proteolytic activity may play a physiological role in the brain.

Adenosine Triphosphate↗

[The change of nerve growth factor and brain derived neurotrophic factor in neurons of cerebral cortex of adult rat following local ischemia].

OBJECTIVE: To acquire knowledge about the change of nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) in neurons of cerebral cortex of adult rat following local ischemia. METHODS: Using specific antiserums of NGF and BDNF by immunohistochemical ABC method. RESULTS: NGF-like and BDNF-like immunoreactions distributed mainly in the neurons of the third and fifth layers in cerebral cortex. After local ischemia, the average gray degrees of NGF and BDNF in neurons of cerebral cortex both decreased on the operated side more than on the un-operated side. CONCLUSION: This experiment demonstrated that the levels of NGF and BDNF in neurons of cerebral cortex following ischemia were upregulated apparently, suggesting that NGF and BDNF may play an important role in the process of neurons' reaction after ischemia.

Animals↗