Stimulation of RNA synthesis in brain cortex of rats after hypoxia by transplantation of embryonic nervous tissue

Our early studies (Polezhaev and Alexandrova, 1986) have shown that acute hypoxic hypoxia in rats causes mass (up to 36%) diffuse dystrophy of brain cortex neurons and that after transplantation of embryonic nervous tissue (ENT) into the brain of hypoxia-exposed rats the dystrophy and death of cortical neurons are reduced to 25% due to normalization of a part of dystrophic neurons. In the present work we studied changes in the RNA synthesis in neurons and in the total brain cortex of rats after hypoxia and subsequent transplantation of ENT into their brain by the autoradiographic and biochemical methods using 14C-adenine. It has been shown that under the action of hypoxia the RNA synthesis in neurons and in the total brain cortex of rats is reduced and after transplantation of ENT into the brain of these rats the RNA synthesis is stimulated and normalized both in neurons and in the total brain cortex.     

大鼠急性局灶性脑挫裂伤后神经细胞线粒体膜电位变化及其意义

目的 研究大鼠急性局灶性脑挫裂伤后神经细胞线粒体膜电位变化及其意义. 方法 70只SD大鼠采用随机数字表法分为假手术组(n=10)和脑损伤组,损伤组按伤后1、6、24、48、72和168 h观察时间点分为6个亚组(n=10).用Feeney氏自由落体撞击法制作重型颅脑损伤模型.利用激光扫描共聚焦显微镜观察损伤后不同时间荧光探剂JC-1标记的活体脑片细胞线粒体膜电位的动态变化,应用Hochest33342荧光染色和Tunel法观察神经细胞凋亡程度,透射电镜观察神经细胞超微结构. 结果 (1)与假手术组相比,损伤组脑皮质神经细胞线粒体膜电位在伤后1 h即开始显著降低.24h降至最低,之后一直无恢复(P<0.01).(2)Hochest33342荧光染色发现神经细胞呈现凋亡的特征.(3)Tunel染色显示损伤组在1 h可见少量阳性细胞,6h阳性细胞数明显增加.48h达峰值,与假手术组比较差异有统计学意义(P<0.05),与线粒体膜电位下降时间比较延迟24 h.(4)电镜发现脑损伤后6h神经细胞核发生染色质散在,核仁边集.细胞核核膜增宽等凋亡特征;24h出现核皱缩,核内染色质溶解. 结论 创伤性脑损伤早期(<1 h)神经细胞线粒体膜电位即下降.并引起神经细胞凋亡.     

Neuronal nitric oxide synthase expression in developing and adult human CNS

Neuronal nitric oxide synthase (nNOS) is constitutively expressed by subpopulations of neurons in the CNS and is involved in neurotransmission, learning and memory, and neuronal injury. While the distribution of nNOS neurons has been characterized in the rodent CNS, the expression in human brain has not been well documented. We determined the expression of nNOS in second trimester human fetal and adult brain. In second trimester fetal brain, the nNOS neurons are concentrated in the developing cerebral cortex at the subplate zone and in layer VI, the striatum, and in certain brainstem nuclei. The nNOS neurons are sparsely distributed in the hippocampus, and virtually absent in the cerebellar cortex. The nNOS neurons in the subplate zone extend their processes radially, suggesting a developmental role, perhaps in guidance. The number and distribution of NADPH diaphorase-positive neurons corresponds to that of the nNOS neurons. While the distribution of nNOS neurons in the adult brain is similar to that found in fetal brain, the overall density is lower in the adult. The highest density of nNOS neurons is found in the striatum followed by the neocortex. A region-specific role for nNOS neurons in human brain and a potential developmental role for nNOS in the cerebral cortex are suggested by these data.     

Space-time correlations of neuronal firing related to memory storage capacity

Most viable theories of memory require some form of synaptic modification dependent on the correlation of pre- and postsynaptic neuronal firing (which we will denote as the Hebb hypothesis). We show here that a possible consequence of this hypothesis is that the storage capacity of a network of highly interconnected neurons is related to the number of synapses and that this implies that the network can be excited into many different time sequence of firing patterns of assemblies of neurons. The important role played by the assembly (as defined by E. R. John) is discussed in detail. A modified Hebb hypothesis is proposed. The crucial experiments to test the model involve the use of two (or more) extracellular microelectrodes to record, simultaneously, the firing activity of several neurons and thus determine the spatial and temporal cross correlations after presenting a mature animal with a variety of stimuli.     

Cellular scaling rules for primate spinal cords

The spinal cord can be considered a major sensorimotor interface between the body and the brain. How does the spinal cord scale with body and brain mass, and how are its numbers of neurons related to the number of neurons in the brain across species of different body and brain sizes? Here we determine the cellular composition of the spinal cord in eight primate species and find that its number of neurons varies as a linear function of cord length, and accompanies body mass raised to an exponent close to 1/3. This relationship suggests that the extension, mass and number of neurons that compose the spinal cord are related to body length, rather than to body mass or surface. Moreover, we show that although brain mass increases linearly with cord mass, the number of neurons in the brain increases with the number of neurons in the spinal cord raised to the power of 1.7. This faster addition of neurons to the brain than to the spinal cord is consistent with current views on how larger brains add complexity to the processing of environmental and somatic information.     

Axonal regeneration of descending brain neurons in larval lamprey demonstrated by retrograde double labeling

In larval lamprey, the large, identified descending brain neurons (Müller and Mauthner cells) are capable of axonal regeneration. However, smaller, unidentified descending brain neurons, such as many of the reticulospinal (RS) neurons, probably initiate locomotion, and it is not known whether the majority of these neurons regenerate their axons after spinal cord transection. In the present study, this issue was addressed by using double labeling of descending brain neurons. In double-label control animals, in which Fluoro-Gold (FG) was applied to the spinal cord at 40% body length (BL; measured from anterior to posterior from tip of head) and Texas red dextran amine (TRDA) was applied later to the spinal cord at 20% BL, an average of 98% of descending brain neurons were double labeled. In double-label experimental animals, FG was applied to the spinal cord at 40% BL; two weeks later the spinal cord was transected at 10% BL; and, eight weeks or 16 weeks after spinal cord transection, TRDA was applied to the spinal cord at 20% BL. At eight weeks and 16 weeks after spinal cord transection, an average of 49% and 68%, respectively, of descending brain neurons, including many unidentified RS neurons, were double labeled. These results in larval lamprey are the first to demonstrate that the majority of descending brain neurons, including small, unidentified RS neurons, regenerate their axons after spinal cord transection. Therefore, in spinal cord-transected lamprey, axonal regeneration of descending brain neurons probably contributes significantly to the recovery of locomotor function.     

Survey of MeCP2 in the Rett syndrome and the non-Rett syndrome brain

The clinical and neuropathologic aspects of Rett syndrome suggest that an arrest of brain development produces the phenotype, but it is not understood how the gene implicated in Rett syndrome, methyl-CpG protein 2 (MeCP2), is regulated during brain development. In this study, the ontogeny of MeCP2 is examined in the developing human brain and in the female Rett syndrome brain to evaluate the relationship between MeCP2 expression and brain development in health and disease, respectively. Immunocytochemistry using an antibody to the C-terminal region of the protein was performed in paraffin sections of the developing brain to define the age and the sites of MeCP2 protein expression. In development, there is no MeCP2 expression in the germinal matrix or in the progenitor cells. At 10 to 14 weeks' gestation, the neurons of the brain stem and the Cajal-Retzius and subplate neurons of the cortex express MeCP2. By midgestation, some neurons of the basal ganglia express MeCP2, and at late gestation, the most mature cortical neurons in the lower cortical layers are positive. The postnatal cortex continues to increase its expression of neuronal MeCP2. In the Rett syndrome brain, fewer neurons express MeCP2 than in the normal brain. This reduction is most apparent in the brain stem and thalamus. The neurons of the cerebral cortex show the least reduction. We conclude that the regulation of MeCP2 abundance is related to human brain development, being expressed in neurons when they appear mature. In Rett syndrome, however, the expression pattern of MeCP2 does not completely resemble that of the normal immature brain, suggesting that the maintenance of MeCP2 might be determined in specific neurons by factors other than those controlling maturation. In the developing brain, synaptic activity and plasticity could be necessary to maintain MeCP2 in selected neuronal populations.     

IgG-immunostaining in the intact rabbit brain: variable but significant staining of hippocampal and cerebellar neurons with anti-IgG

A significant number of brain neurons in the rabbit brain were immunostained with anti-rabbit gamma-immunoglobulin (IgG). IgG-positive neurons were often found in the cerebellum, lower brainstem and motor nuclei. Similar IgG-positive neurons were occasionally found in the hippocampus, cerebral cortex and midbrain, but not in the striatum and thalamus. These neurons showed very clear Golgi-like staining of soma and dendrites but IgG staining was absent from the cell nuclei and axons. In particular, groups of Purkinje neurons in the rabbit cerebellum showed strong IgG-positive staining. To confirm whether the staining reflected the existence of IgG molecules in these neurons, staining specificity was carefully evaluated. Staining was specifically eliminated by pre-absorption of the antibodies with the purified rabbit IgG. An antibody to the neural cell adhesion molecule (NCAM or CD56), a member of the immunoglobulin superfamily, exhibited a completely different pattern of staining as that for IgG. To determine whether IgG-like immunoreactivity was a general feature of mammalian brain, brain sections of rabbits, rats, and mice were immunostained with antibodies to IgGs of each of the three species. Similar IgG-positive neurons were observed in all three species, although the distribution and frequency was characteristic for each species. In rabbit brain, anti-rabbit IgG stained-neurons were more abundant compared to rat and mouse brain. IgG-positive microglia-like cells were evident in mouse brain, but less frequent in rabbit and were hardly observed in rat brain. To evaluate whether stained neurons could synthesize IgG, in situ hybridization was carried out using an antisense oligonucleotide probe to rabbit IgG DNA. No significant label was observed in cerebellum. These results suggest that a significant number of neurons in the intact rabbit brain take up IgGs and concentrate them in their cytoplasm, although the molecular uptake mechanism is retained for future studies. Our results also suggest that the rabbit may be a suitable animal to study the function(s) of IgG in brain neurons.     

Distribution of parvalbumin immunoreactivity in the human brain

Summary The cellular distribution of parvalbumin-like immunoreactivity (PA-LI) in the human brain was investigated by peroxidase-antiperoxidase methods using antiserum to rat skeletal muscle parvalbumin. PA-LI was present in non-pyramidal neurons of the cerebral cortices, stellate cells, basket cells and Purkinje cells in cerebellar cortices, and neurons of some nuclei in human brain stem; the distribution of PA-LI in human brain was very similar to that in rat brain. These results indicate that PA-LI is widely distributed in a specific subpopulation of neurons of the human brain.     

Basolateral amygdala regulation of adult hippocampal neurogenesis and fear-related activation of newborn neurons

Impaired regulation of emotional memory is a feature of several affective disorders, including depression, anxiety and post-traumatic stress disorder. Such regulation occurs, in part, by interactions between the hippocampus and the basolateral amygdala (BLA). Recent studies have indicated that within the adult hippocampus, newborn neurons may contribute to support emotional memory, and that regulation of hippocampal neurogenesis is implicated in depressive disorders. How emotional information affects newborn neurons in adults is not clear. Given the role of the BLA in hippocampus-dependent emotional memory, we investigated whether hippocampal neurogenesis was sensitive to emotional stimuli from the BLA. We show that BLA lesions suppress adult neurogenesis, while lesions of the central nucleus of the amygdala do not. Similarly, we show that reducing BLA activity through viral vector-mediated overexpression of an outwardly rectifying potassium channel suppresses neurogenesis. We also show that BLA lesions prevent selective activation of immature newborn neurons in response to a fear-conditioning task. These results demonstrate that BLA activity regulates adult hippocampal neurogenesis and the fear context-specific activation of newborn neurons. Together, these findings denote functional implications for proliferation and recruitment of new neurons into emotional memory circuits.     

Equal numbers of neuronal and nonneuronal cells make the human brain an isometrically scaled‐up primate brain

The human brain is often considered to be the most cognitively capable among mammalian brains and to be much larger than expected for a mammal of our body size. Although the number of neurons is generally assumed to be a determinant of computational power, and despite the widespread quotes that the human brain contains 100 billion neurons and ten times more glial cells, the absolute number of neurons and glial cells in the human brain remains unknown. Here we determine these numbers by using the isotropic fractionator and compare them with the expected values for a human‐sized primate. We find that the adult male human brain contains on average 86.1 ± 8.1 billion NeuN‐positive cells (“neurons”) and 84.6 ± 9.8 billion NeuN‐negative (“nonneuronal”) cells. With only 19% of all neurons located in the cerebral cortex, greater cortical size (representing 82% of total brain mass) in humans compared with other primates does not reflect an increased relative number of cortical neurons. The ratios between glial cells and neurons in the human brain structures are similar to those found in other primates, and their numbers of cells match those expected for a primate of human proportions. These findings challenge the common view that humans stand out from other primates in their brain composition and indicate that, with regard to numbers of neuronal and nonneuronal cells, the human brain is an isometrically scaled‐up primate brain. J. Comp. Neurol. 513:532–541, 2009. © 2009 Wiley‐Liss, Inc.     

Clusterin (SGP-2): A multifunctional glycoprotein with regional expression in astrocytes and neurons of the adult rat brain

Clusterin (SGP-2) is a newly described glycoprotein associated with several putative functions including responses to brain injury. This study reports the regional and cell type expression of clusterin mRNA and its encoded glycoprotein in the rat brain; a limited comparison was also done with the human brain. Using in situ hybridization combined with immunocytochemistry, we found that astrocytes and neurons may express clusterin mRNA in the normal adult brain. While astrocytes throughout the brain contained clusterin mRNA, there was regional selectivity for neuronal clusterin expression. In the striatum, clusterin mRNA was not detected in neurons. Only a subset of substantia nigra dopaminergic neurons or locus ceruleus noradrenergic neurons (tyrosine hydroxylase immunopositive) contained clusterin mRNA. However, neuronal clusterin mRNA was prevaìent in pontine nuclei and in the red nucleus of the midbrain tegmentum. Similarly, clusterin mRNA was prevalent in both rat and human hippocampal neuron-specific enolase immunopositive pyramidal neurons, although rat CA1 neurons had less mRNA than CA2–CA3 neurons. Monotypic primary cell cultures from the neonatal rat showed clusterin mRNA in both neurons and astrocytes, but not in microglia. By immunocytochemistry, no clusterin immunopositive glia were observed in any region of the rat brain, confirming previous studies. However, clusterin immunopositive cells (putative neurons) were observed in the Purkinje cell layer of the cerebellum, medial and interposed cerebellar nuclei, trigeminal motor nucleus, and red nucleus. Finally, in vitro studies suggest that astrocytes, but not neurons, secrete clusterin, which is pertinent to clusterin immunodeposits found after experimental lesioning. © 1994 Wiley-Liss, Inc.     

Localization of neuregulin-1alpha (heregulin-alpha) and one of its receptors, ErbB-4 tyrosine kinase, in developing and adult human brain

Using immunohistochemistry, Western blot analysis, and RT-polymerase chain reaction, we studied the distribution of neuregulin-1 splice variant alpha (NRG-1alpha) and one of its putative receptors, ErbB-4 tyrosine kinase, in human brain. In the pre- and perinatal human brain immunoreactivity was confined to numerous neurons, with the highest cell density found in cortical gray matter, hypothalamus and cerebellum. In the adult brain, single cortical gray and white matter neurons showed NRG-1alpha immunoreactivity. Occasionally, immunoreactive oligodendrocytes were observed. NRG-1alpha-expressing neurons were also found in the hypothalamus, hippocampus, basal ganglia and brain stem. Application of two antibodies recognizing alpha and beta isoforms revealed a different distribution pattern in that many cortical and hippocampal pyramidal neurons were labeled. ErbB-4 immunoreactivity was expressed in both neurons and oligodendrocytes. Our data show that NRG-1alpha expression is lower in the adult human brain than in the developing brain, and, therefore, support a role for NRG-1alpha in brain development.     

Wandering neuronal migration in the postnatal vertebrate forebrain

Most non-mammalian vertebrate species add new neurons to existing brain circuits throughout life, a process thought to be essential for tissue maintenance, repair, and learning. How these new neurons migrate through the mature brain and which cues trigger their integration within a functioning circuit is not known. To address these questions, we used two-photon microscopy to image the addition of genetically labeled newly generated neurons into the brain of juvenile zebra finches. Time-lapse in vivo imaging revealed that the majority of migratory new neurons exhibited a multipolar morphology and moved in a nonlinear manner for hundreds of micrometers. Young neurons did not use radial glia or blood vessels as a migratory scaffold; instead, cells extended several motile processes in different directions and moved by somal translocation along an existing process. Neurons were observed migrating for ～2 weeks after labeling injection. New neurons were observed to integrate in close proximity to the soma of mature neurons, a behavior that may explain the emergence of clusters of neuronal cell bodies in the adult songbird brain. These results provide direct, in vivo evidence for a wandering form of neuronal migration involved in the addition of new neurons in the postnatal brain.     

Electroshock after discharges are related to neuronal oscillability: Neuronal models of Aplysia

Identifiable giant neurons of Aplysia explored intracellularly behave differently at the offset of an intracellular electroshock (IES) or after a synaptic 'tetanization', according to their functional type: neurons of the stable type depolarize and fire at the offset of the IES, anodal or cathodal, thus eliciting an afterdischarge (AD). The threshold of this AD is lowered if the neuron is destabilized, i.e. converted from the stable to an oscillatory type (for instance by decalcification). Neurons normally of the tonic type are more sensitive to an IES, eliciting a longer afterdischarge than the stable neurons. Extracellular electroshock (EES) anodal or cathodal, applied directly on desheathed somata of Helix give long-lasting afterdischarges at the offset. In addition, EES stimulating presynaptic terminals or axons leads to a high frequency synaptic input on remote neurons. At the offset of this input either prolonged synaptic afterdischarges or postsynaptic rebounds of the membrane potential sustaining bursts of decreasing amplitude denote apparent oscillatory properties of the synaptically activated neuron. Finally, any conversion by convulsants of tonic neurons to oscillators highly facilitates the elicitation of afterdischarges of axons simultaneous to paroxysmal depolarization shifts of the homologous somata. These results indicate that afterdischarge elicitation is highly facilitated (low threshold) in normal oscillatory neurons and/or chemically destabilized neurons.     

Primary culture of neurons derived from the adult mammalian brain.

A method is described for the isolation and culture of neurons in the adult mammalian brain. The cell could be maintained in primary culture for more than several weeks. Whereas the neurons freshly dissociated from the adult brain did not respond to any of the neurotransmitter substances applied, the neurons regained the ability to respond to a variety of neurotransmitters when cultured. The cultured neurons of the adult mammalian brain may be an excellent model for physiological as well as pharmacological investigations of the central nervous system.     

Descending brain neurons in larval lamprey: Spinal projection patterns and initiation of locomotion

In larval lamprey, partial lesions were made in the rostral spinal cord to determine which spinal tracts are important for descending activation of locomotion and to identify descending brain neurons that project in these tracts. In whole animals and in vitro brain/spinal cord preparations, brain-initiated spinal locomotor activity was present when the lateral or intermediate spinal tracts were spared but usually was abolished when the medial tracts were spared. We previously showed that descending brain neurons are located in eleven cell groups, including reticulospinal (RS) neurons in the mesenecephalic reticular nucleus (MRN) as well as the anterior (ARRN), middle (MRRN), and posterior (PRRN) rhombencephalic reticular nuclei. Other descending brain neurons are located in the diencephalic (Di) as well as the anterolateral (ALV), dorsolateral (DLV), and posterolateral (PLV) vagal groups. In the present study, the Mauthner and auxillary Mauthner cells, most neurons in the Di, ALV, DLV, and PLV cell groups, and some neurons in the ARRN and PRRN had crossed descending axons. The majority of neurons projecting in medial spinal tracts included large identified Müller cells and neurons in the Di, MRN, ALV, and DLV. Axons of individual descending brain neurons usually did not switch spinal tracts, have branches in multiple tracts, or cross the midline within the rostral cord. Most neurons that projected in the lateral/intermediate spinal tracts were in the ARRN, MRRN, and PRRN. Thus, output neurons of the locomotor command system are distributed in several reticular nuclei, whose neurons project in relatively wide areas of the cord.     

Morphological, biochemical and physiological changes in brain nervous tissue of adult intact and hypoxia-subjected rats after transplantation of embryonic nervous tissue

Data of morphological, biochemical and physiological investigations of brain nervous tissue in adult intact and hypoxia-subjected rats after transplantation of rat embryonic nervous tissue are presented. It has been established that transplants survive and develop successfully in the brain of both intact and hypoxia-subjected rats. The Golgi technique reveals that transplanted neurons form dendrites and axons. They also establish afferent and efferent connections with neurons in different brain regions of the recipient, as has been demonstrated by the horseradish peroxidase (HRP) tract tracing technique. Hypoxia induces reversible as well as irreversible dystrophy of host brain neurons, and transplantation of embryonic brain tissue leads to considerable normalization of the state of reversibly dystrophic neurons. Biochemical investigations have revealed that the spectrum of cytoplasmic proteins of neurons and proteins of the whole brain cortex tissue changed after hypoxia are significantly normalized after subsequent transplantation. It should be also pointed out that there is a considerable increase in the number of proteins in certain fractions which is correlated with regeneration of reversibly dystrophic neurons and other cortical cells. Electrophysiological experiments revealed distinct but short-term EEG changes induced by hypoxia. 1-2 days after hypoxia all EEG parameters were back to normal and displayed no changes during the 2-3-month registration period.     

Immunohistochemical localization of hyaluronic acid in rat and human brain

The immunohistochemical localization of hyaluronic acid (HA) was studied in rat and human brain using the monoclonal antibody NDOG1, which specifically recognizes HA. In both rat and human brain, HA-like immunoreactivity formed characteristic coats around neurons in highly selective areas. The staining was abolished by pretreatment of sections with testicular andStreptomyces hyaluronidases, indicating that the staining was specific for HA. In rat brain, positive neurons were located in the cerebral cortex, subiculum, amygdala, thalamic reticular nucleus, nuclei of the inferior colliculus, nuclei of the trapezoid body, and vestibular nuclei. They were also scattered in the hypothalamus, substantia nigra pars reticularis, red nucleus, parabrachial nuclei, brainstem reticular nuclear group, ventral cochlear nucleus, nuclei of lateral lemniscus, and deep cerebellar nuclei. Double immunohistochemical studies showed that many neurons staining for HA were positive for parvalbumin, with minor exceptions in the amygdala and piriform cortex, where some HA-positive neurons were also positive for calbindin-D28k. In the areas studied in human brain, the distribution of HA-positive neurons was virtually identical to that in rat brain. HA-positive neurons were not significantly altered in Alzheimer disease (AD) brain, suggesting that these neurons are resistant to the pathological process of AD.