NADPH-diaphorase activity in the superficial layers of the superior colliculus of rats during aging

Neurons in the superficial layers of the superior colliculus are key elements in the visual system of rodents since they receive extensive afferent projections from retinal ganglion cells. The NADPH-diaphorase histochemical technique was used to detect differences in neuronal nitric oxide synthase (nNOS) in the superficial layers of the superior colliculus (sSC) of young adult (3 months) and aged (24 and 26 months) rats. The orientation of the dendritic processes of NADPH-diaphorase-positive neurons, cross-sectional area, and number of neurons per mm2 were analyzed. NADPH-d histochemistry revealed a high number of NADPH-d-positive cells in the stratum zonale and stratum griseum superficiale in adult and aged animals. NADPH-d-positive neurons were classified into the following morphological types: marginal, horizontal, pyriform, narrow-field vertical, wide-field vertical, and stellate. During aging, narrow field vertical and wide field vertical neurons present somatic atrophy and an increase in dendritic processes with dorsoventral orientation, whereas wide field vertical neurons show a decrease in those with lateromedial orientation. Marginal neurons undergo somatic hypertrophy at 26 months when compared with those at 3 months. The remaining types of neurons do not undergo size changes. Finally, the number of NADPH-d-positive neurons per mm2 in the various types of morphology does not significantly change with age. It is suggested to be likely that the aging process in the nitrergic neurons of the sSC does not lead to significant changes in the synthesis of NO from the constitutive NOS isoforms.     

Distribution of nitric oxide synthase immunoreactivity in the mouse brain

Nitric oxide (NO) is a gaseous intercellular messenger with a wide range of neural functions. NO is synthesized by activation of different isoforms of nitric oxide synthases (NOS). At present NOS immunoreactivity has been described in mouse brain in restricted and definite areas and no detailed mapping studies have yet been reported for NOS immunoreactivity. We have studied the distribution of neuronal NOS-containing neurons in the brain of three months male mice, using a specific commercial polyclonal antibody against the neuronal isoform of nitric oxide synthase (nNOS). Neuronal cell bodies exhibiting nNOS immunoreactivity were found in several distinct nuclei throughout the brain. The neurons that were positively stained exhibited different intensities of reaction. In some brain areas (i.e., cortex, striatum, tegmental nuclei) neurons were intensely stained in a Golgi-like fashion. In other regions, immunoreactive cells are moderately stained (i.e., magnocellular nucleus of the posterior commissure, amygdaloid nucleus, interpeduncular nucleus, lateral periaqueductal gray) or weakly stained (i.e., vascular organ of the lamina terminalis, hippocampus, inferior colliculus, reticular nucleus). In the mouse, the NO-producing system appears well developed and widely diffused. In particular, nNOS immunoreactive neurons seem chiefly present in several sensory pathways like all the nuclei of the olfactory system, as well as in many regions of the lymbic system. These data suggest a widespread role for the NO system in the mouse nervous system.     

Functional neuroimaging of memory: implications for cognitive aging

Our understanding of the ways in which changes in specific neural systems mediate adult age differences in memory is rapidly increasing, due in no small part to the advent of functional neuroimaging techniques. This article reviews age-related changes in memory performance obtained with behavioral measures, describes models of the neural mechanisms of memory, and derives predictions from these models regarding age-related changes in brain activation patterns. The neuroimaging findings obtained to date support models emphasizing the role of prefrontal cortex in age-related changes in memory functioning, especially for episodic memory retrieval. In general, neural activation associated with episodic memory encoding is regionally similar for younger and older adults but relatively lower in magnitude for older adults. During retrieval, activation that is restricted to the right prefrontal cortex for younger adults is more likely to be bilateral for older adults. Prefrontal activation exhibits an age-related increase when working memory tasks require simple storage and an age-related decrease when working memory requires higher-level executive processes. Although the evidence is limited, behavioral performance and activation patterns appear to be similar among younger and older adults on tests of semantic (context-independent) and implicit memory. We conclude that several methodological issues, such as defining the relation between brain structure and function, and determining the relationship between performance and activation, are particularly important for understanding age-related changes. Future directions for aging research include further investigation of the relation between encoding and retrieval and the identification of both spared and impaired neural systems.     

基于听觉神经同步振荡网络的时频结构描述方法

人类听觉系统具有优良的非平稳信号分析能力,在听觉系统中,由耳蜗基底膜对信号进行类似于带通滤波的时频分解,并由内毛细胞、传入神经和听觉中枢的神经网络对时频分解结果逐步进行特征信息提取和压缩。鉴于此,参照Wang-Brown模型,建立一种可描述信号时频结构特征的听觉模型,该模型包括基底膜、内毛细胞、中级听觉和听觉中枢等子模型,听觉中枢模型由单层听神经振荡网络构成。略去Wang-Brown模型中随机项和侧抑制项,简化内毛细胞模型,设计听神经元的活跃准则和神经元间的联接方式。信号经基底膜、内毛细胞和中级听觉模型处理后,由听神经振荡网络进行信息综合,使得信号中时频结构相似的区域所对应的听神经元进行同步振荡,从而可利用同步振荡神经元的分布情况描述信号的时频结构。进行故障转子升降速试验和风力发电增速机稳速运行试验,试验所得信号的分析结果表明,所建模型能够有效描述信号的时频结构特征及其变化情况,对信号的瞬态变化较为敏感,且数据量相对较小,易于智能识别。     

Stereological methods in the analysis of neuronal parameters in the central nervous system*

The structure of the central nervous system (CNS) is briefly explained. It is compared with the electronic computer and the differences between the two are stated. The methods of measuring this structure and its function are reported. The surface of the cortex as well as the volume of different grey and white matters making up the brain can macroscopically be evaluated on serial slices with the aid of stereological procedures. There is an exponential correlation between volume of the cortex and volume of brain and body in mammals (Fig. 3). The density of neurons and the lengths of their processes can be examined by light microscopy. An improved procedure of counting neurons in the cortex is described. With regard to mammals it is interesting that the densities of neurons in larger brains are lower than in smaller ones. The correlation is negatively exponential. Man and primates have a higher density of cells than the rest of the mammals; their neurons, however, are small. This corresponds to a specialization that can be considered to represent a miniaturization of the neurons (Fig. 4). The topological analysis of dendritic trees is mentioned. The lengths, surfaces and volumes of processes of cells and the amount of synapses can be measured on electron micrographs. The principles and problems of stereology are described by an example of how to measure the myelinated fibres. Some results of the development of these fibres from birth on are reported upon (Fig. 6). In conclusion, some data of the probable dimensions of the human brain are given.     

Immunohistochemistry of neuronal nitric oxide synthase and protein nitration in the striatum of the aged rat

To ascertain the possible implications of the nitric oxide (NO*) producing system in striatal senescence, and by using immunohistochemistry and image-processing approaches, we describe the presence of the enzyme nitric oxide synthase (NOS), the NADPH-diaphorase (NADPH-d) histochemical marker, and nitrotyrosine-derived complexes (N-Tyr) in the striatum of adult and aged rats. The results showed neuronal NOS immunoreactive (nNOS-IR) aspiny medium-sized neurons and nervous fibres in both age groups, with no variation in the percentage of immunoreactive area but a significant decrease in the intensity and in the number of somata with age, which were not related to the observed increase with age of the striatal bundles of the white matter. In addition, NADPH-d activity was detected in neurons with morphology similar to that of the nNOS-IR cells; a decrease in the percentage of area per field and in the number of cells, but an increase in the intensity of staining for the NADPH-d histochemical marker, were detected with age. The number of neuronal NADPH-d somata was higher than for the nNOS-IR ones in both age groups. Moreover, N-Tyr-IR complexes were observed in cells (neurons and glia) and fibres, with a significant increase in the percentage of the area of immunoreaction, related to the increase of white matter, but a decrease in intensity for the aged group. On the other hand, we did not detect the inducible isoform (iNOS) either in adult or in aged rats. Taken together, these results support the contention that NADPH-d staining is not such an unambiguous marker for nNOS, and that increased protein nitration may participate in striatal aging.     

Microscopic study of certain age-related structural changes of maxillary sinus lining epithelium in albino rats

Mucociliary clearance is essential to maintain the defense function of the maxillary sinus; however, no literatures described the age changes in its lining epithelium. Therefore, the current work sought to describe the morphological postnatal age-related changes of maxillary sinus lining epithelium in rats using light, transmission, and scanning electron microscopes. Eighteen albino rats were divided into six groups according to their ages: 2-week-old, 1-month-old, 2-month-old, 3-month-old, adults, and senile rats. One-month-old-rats' group was the first to have recognizable maxillary sinus cavities that were lined by either single flat cellular layer or two distinct epithelial layers. These cells were devoid of microvilli and cilia, none of them showed evidence of differentiation into identifiable cell types. In 2- and 3-month-old rats, the mucosa of maxillary sinus started to be lined with pseudostratified epithelium with apparent increase in both microvilli and cilia. The first indication of goblet cell differentiation was observed in 3-month-old-rats. In the adult rats, the sinuses became completely lined by mature respiratory epithelium. However, in senile rats the epithelium exhibited polyps with clumped cilia and some areas of stratification and desquamation. Goblet cells were scanty and degenerating. The impaired mucociliary components (epithelium, cilia, and goblet cells' mucus) found in young and old ages of the current work might be correlated to human to explain predisposition of rhino-sinusitis in these age groups.     

Supraspinal connections of the ovary: structural and functional aspects

This review summarizes our recent studies using the viral transneuronal tracing technique to identify sites in the central nervous system (CNS) that are connected with the ovary. A neurotropic virus (pseudorabies virus) was injected into the ovary and various times after the inoculation the spinal cord and brain were examined for virus-infected neurons identified by immunocytochemistry. Such neurons could be detected in well-defined cell groups of the spinal cord (intermediolateral cell column), brain stem (vagal nuclei, area postrema, parapyramidal nucleus, caudal raphe nuclei, A1, A5, A7 noradrenergic cell groups, locus coeruleus, Barrington's nucleus, periaqueductal gray), hypothalamus (paraventricular nucleus, anterior hypothalamus, arcuate nucleus, zona incerta), and, at longer survival time, in some telencephalic structures (amygdala, bed nucleus of the stria terminalis). These findings provided the first neuromorphological evidence for the existence of a multisynaptic neuronal pathway between the brain and the ovary presumably involved in the neuronal control of the organ. The observations indicate that there is a significant overlap of CNS structures connected with the ovary, the testis, other organs and organ systems, suggesting similar neuronal circuitries of the autonomic nervous system innervating the different organs. The known descending neuronal connections between the CNS structures labeled from the ovary by the viral transneuronal tracing technique and the findings suggesting a pituitary independent interplay between certain cerebral structures such as the hypothalamus, the amygdala, and the ovary are also summarized in this review.     

Afferent projections of the superior olivary complex

Based on current literature, the afferents of the superior olivary complex (SOC) are described including those from the cochlear nucleus, inferior colliculus, thalamus, and auditory cortex. Intrinsic SOC afferents and non-auditory afferents from the serotoninergic and noradrenergic systems are also described. New data are provided that show a differential distribution of serotoninergic afferents within the SOC: serotoninergic fibers were relatively sparse in the lateral and medial superior olives and the medial nucleus of the trapezoid body and were most numerous in periolivary regions. There are variations in the density of serotoninergic fibers within periolivary regions themselves. New data is also provided on auditory and non-auditory afferents to SOC neurons, which have known targets. These include: cochlear nucleus afferents to periolivary (lateral nucleus of the trapezoid body, LNTB) cells that project to the inferior colliculus; cortical afferents to periolivary (ventral nucleus of the trapezoid body, VNTB) cells that project to the cochlear nucleus; and serotoninergic and noradrenergic afferents to periolivary (LNTB and VNTB) cells that project to the cochlear nucleus. The relationships between other types of afferents and SOC neurons with known projections are also described as functional circuits. The circuits include those that are part of the ascending auditory system (to the inferior and superior colliculi, lateral lemniscus, and medial geniculate nucleus), the descending auditory system (to the cochlea and cochlear nucleus), and the middle ear reflex circuits.     

Neurotrophins and their receptors in the primary olfactory neuraxis

The primary olfactory pathway is an elegant and simple system in which to study neurogenesis and neuronal plasticity because of the simple fact that olfactory receptor neurons (ORNs) are continually generated throughout the adult lifetimes of vertebrates. Thus, neuronal birth, differentiation, survival, axon pathfinding, target recognition, synapse formation, and cell death are developmental events that can be examined in the mature olfactory epithelium (OE). Neurotrophins (nerve growth factor, brain-derived neurotrophic factor, and neurotrophin 3, and 4/5) are a family of bioactive peptides that exert their effects by interacting with high- and low-affinity receptors on the surfaces of responsive cells, and have been implicated in several stages of neuronal development throughout the central and peripheral nervous system (CNS and PNS). There has been significant interest within the olfactory community as to how these multifunctional peptides might regulate the cycle of degeneration and regeneration of olfactory receptor neurons. The focus of this review is to highlight what is known about the actions of neurotrophins in the primary olfactory pathway, and to pinpoint future directions that will enable us to further understand their role in olfactory receptor neuron development and turnover.     

Neuroanatomical and cognitive correlates of adult age differences in acquisition of a perceptual-motor skill

The objective of this study was to examine age differences in procedural learning and performance in conjunction with differential aging of central nervous system (CNS) structures. Sixty-eight healthy volunteers (age 22-80) performed a pursuit rotor task (four blocks of 20 15-second trials each). Volumes of the cerebellar hemispheres, neostriatum, prefrontal cortex, and hippocampus were measured from Magnetic Resonance (MR) images. Improvement in pursuit rotor performance was indexed by increase in time on target (TOT). A general improvement trend was evident across the blocks of trials. Overall, younger participants showed significantly longer TOT. The rate of improvement was age-invariant during the initial stages of skill acquisition but became greater in middle-aged participants as the practice progressed. When the influences of regional brain volumes were taken into account, the direct age effect on mean TOT measured during the first day of practice disappeared. Instead, reduced volumes of the cerebellar hemispheres and the putamen and poorer performance on nonverbal working memory tasks predicted shorter TOT. In contrast, neither the volume of the caudate and the hippocampus, nor verbal working memory showed association with motor performance. Pursuit rotor performance at the later stages of practice was unrelated to the reduction in putamen volume and was affected directly by age, cerebellar volume, and nonverbal working memory proficiency. We conclude that in a healthy population showing no clinical signs of extrapyramidal disease, age-related declines in procedural learning are associated with reduced volume of the cerebellar hemispheres and lower nonverbal working memory scores. During initial stages of skill acquisition, reduced volume of the putamen is also predictive of poorer performance.     

Comparison of transsynaptic viral labeling of central nervous system structures from the uterine horn in virgin, pregnant, and lactating rats

Using the transneuronal viral tracing method, the central nervous system (CNS) connections of the uterine horn were studied in virgin, pregnant, and in lactating rats. The frequency of viral labeling in the brain and the distribution of virus-infected neurons from the uterine horn were compared among groups. There was a marked difference in the frequency of viral labeling in the brain stem. In virgin rats more than half of the brain stems (5 out of 9) were labeled. In contrast, in pregnant animals viral-labeled neurons were detected in only a few cases (3 out of 16) and almost each brain stem of the lactating group was labeled (12 out of 13). A similar, less marked difference was observed in the hypothalamus. The pattern of distribution of infected neurons was similar in each group. In the brain stem, the nucleus of the solitary tract, dorsal motor nucleus of the vagus, area postrema, gigantocellular and paragigantocellular nucleus, ventrolateral medulla, A5 cell group, and caudal raphe nuclei were the most frequently labeled structures. In the diencephalon, viral-infected neurons were detected primarily in the hypothalamic paraventricular nucleus. The telencephalon was devoid of infected cells. Data suggest that the CNS control of the uterine horn varies depending on reproductive status. The low frequency of brain labeling in pregnant rats may be related to the almost complete lack of sympathetic fibers in the uterus prior to parturition and the very high frequency of labeling in lactating animals to the postpartum hyperinnervation of the uterus.     

Adrenomedullin in the central nervous system

Adrenomedullin (AM) is a novel vasodilator peptide first purified from human pheochromocytoma by tracing its capacity to stimulate cAMP production in platelets. AM immunoreactivity is widely distributed in the central nervous system (CNS) and in the rat has been demonstrated by immunohistochemical techniques to be present in many neurons throughout the brain and spinal cord, as well as in some vascular endothelial cells and perivascular glial cells. Electron microscopy shows that the immunoreactivity is located mainly in the neuronal cytoplasm, but also occurs in the cell nucleus in some cells of the caudate putamen and olfactory tubercle. Biochemical analyses suggest that higher molecular forms, presumably precursor forms, may predominate over fully processed AM in some brain areas. The expression of AM immunoreactivity is increased in cortical neurons, endothelial cells, and perivascular processes after a simulation of ischemia by oxygen and glucose deprivation. Immunohistochemical, electrophysiological, and pharmacological studies suggest that AM in the CNS can act as a neurotransmitter, neuromodulator, or neurohormone, or as a cytoprotective factor in ischemic/hypoxic conditions, in addition to its vasodilator role.     

Distribution and projections of nitric oxide synthase neurons in the rodent superior olivary complex

The superior olivary complex (SOC), a group of interrelated brainstem nuclei, sends efferents to a variety of neuronal structures including the cochlea and the inferior colliculus. The present review describes data obtained from rodents providing evidence that the gaseous, short-living neuroactive substance nitric oxide (NO) is produced in the SOC. The NO-synthesizing enzyme neuronal NO-synthase (nNOS) has been localized by means of several methods including histochemistry and immunohistochemistry. Perikarya containing nNOS were found in several nuclei of the SOC. Their largest numbers and percentages of total cells were observed in the medial nucleus of the trapezoid body. Stained terminals were observed mainly in the lateral superior olivary nucleus and in the superior paraolivary nucleus. While retrograde neuronal tracing identified a considerable number of nNOS-immunoreactive neurons as to be part of the olivo-cochlear pathway, the projection patterns of other nNOS-immunoreactive SOC cell groups remain to be investigated. We also review other putative sources of cochlear NO, and discuss the possible role of NO in the lower auditory brainstem and organ of Corti with regard to physiological and pathophysiological mechanisms.     

Anterograde and retrograde tracing with high molecular weight biotinylated dextran amine through thalamocortical and corticothalamic pathways

Biotinylated dextran amine (BDA) has been used for neural pathway tracing in the central nervous system for many decades, in which high molecular weight BDA appeared to be transported predominantly in the anterograde direction and less in the retrograde direction. In the current study, we reexamined the properties of neural labeling with high molecular weight BDA through a reciprocal neural pathway between thalamus and somatosensory cortex. After injection of BDA into the ventral posteromedial nucleus of thalamus (VPM) in the rat, the BDA labeling was sequentially examined on somatosensory cortex at 3, 5, 7, 10, and 14 survival days. Both of anterogradely labeled axonal terminals and retrogradely labeled neuronal cell bodies were observed simultaneously on the somatosensory cortex. With the increasing of survival times after injection, morphological changes occurred on the labeled axonal arbors and neuronal dendrites, in which the high quality of BDA labeling appeared on the tenth survival day. These results indicate that high molecular weight BDA is not only a sensitive anterograde tracer but also an excellent retrograde marker to be used for tracing through thalamocortical and corticothalamic pathways. And the detailed structure of neural labeling with BDA similar to Golgi‐like resolution can be obtained at optimal survival times of animals after the injection of high molecular weight BDA.     

Distribution of histamine in the CNS of different spiders

Immunohistochemistry is used to demonstrate histamine-immunoreactivity in the CNS of spiders. We found histamine-immunoreactivity in the photoreceptors of different spiders. Therefore, we suggest that histamine is a neurotransmitter of photoreceptors in all arthropods, since it is also known to occur in the photoreceptors of the other main arthropod taxa (Merostomata, Crustacea, and Insecta). We also describe a system of only six omnisegmental histamine-immunoreactive neurons within the central nervous system. These histamine-immunoreactive neurons can be divided into two subgroups: a dorsal system with two cells per hemisphere and a ventral system with only one cell per hemisphere. All six cells have extended arborizations in both the motor and the sensory areas of all neuromeres in the suboesophageal ganglionic mass. In contrast to araneomorph spiders, two additional sets of histamine-immunoreactive neurons were detected in mygalomorph spiders. The first set consists of seventeen cells with their cell bodies located in the cheliceral ganglion and projecting to central areas of the protocerebrum. The second set contains many if not all sensory projections from the tarsal organs on all eight legs and the pedipalps to the Blumenthal neuropil.     

Involvement of the choroid plexus in central nervous system inflammation

During inflammatory conditions in the central nervous system (CNS), immune cells immigrate into the CNS and can be detected in the CNS parenchyma and in the cerebrospinal fluid (CSF). The most comprehensively investigated model for CNS inflammation is experimental autoimmune encephalomyelitis (EAE), which is considered the prototype model for the human disease multiple sclerosis (MS). In EAE autoagressive CD4(+), T cells gain access to the CNS and initiate the molecular and cellular events leading to edema, inflammation, and demyelination in the CNS. The endothelial blood-brain barrier (BBB) has been considered the obvious place of entry for the circulating immune cells into the CNS. A role of the choroid plexus in the pathogenesis of EAE or MS, i.e., as an alternative entry site for circulating lymphocytes directly into the CSF, has not been seriously considered before. However, during EAE, we observed massive ultrastructural changes within the choroid plexus, which are different from changes observed during hypoxia. Using immunohistochemistry and in situ hybridization, we observed expression of VCAM-1 and ICAM-1 in the choroid plexus and demonstrated their upregulation and also de novo expression of MAdCAM-1 during EAE. Ultrastructural studies revealed polar localization of ICAM-1, VCAM-1, and MAdCAM-1 on the apical surface of choroid plexus epithelial cells and their complete absence on the fenestrated endothelial cells within the choroid plexus parenchyme. Furthermore, ICAM-1, VCAM-1, and MAdCAM-1 expressed in choroid plexus epithelium mediated binding of lymphocytes via their known ligands. In vitro, choroid plexus epithelial cells can be induced to express ICAM-1, VCAM-1, MAdCAM-1, and, additionally, MHC class I and II molecules on their surface. Taken together, our observations imply a previously unappreciated function of the choroid plexus in the immunosurveillance of the CNS.     

Giant identified NO-releasing neurons and comparative histochemistry of putative nitrergic systems in gastropod molluscs

Gastropod molluscs provide attractive model systems for behavioral and cellular analyses of the action of nitric oxide (NO), specifically due to the presence of many relatively giant identified nitrergic neurons in their CNS. This paper reviews the data relating to the presence and distribution of NO as well as its synthetic enzyme NO synthase (NOS) in the CNS and peripheral tissues in ecologically and systematically different genera representing main groups of gastropod molluscs. Several species (Lymnaea, Pleurobranchaea, and Aplysia) have been analyzed in greater detail with respect to immunohistochemical, biochemical, biophysical, and physiological studies to further clarify the functional role of NO in these animals.     

Applications of confocal microscopy to the study of myelin development and neuron structure

Confocal laser scanning microscopy has been used to study the localization of myelin basic proteins expressed in nonglial cells, and to probe the three-dimensional structure of central auditory neurons in the lateral superior olive. The paper focuses on the techniques used to obtain the results. The key roles of confocal microscopy and computer image processing of the images obtained are emphasized as they relate to the discovery of essential structural information about these specimens.     

Expression and distribution of S100 protein in the nervous system of the adult zebrafish (Danio rerio)

S100 proteins are EF-hand calcium-binding protein highly preserved during evolution present in both neuronal and non-neuronal tissues of the higher vertebrates. Data about the expression of S100 protein in fishes are scarce, and no data are available on zebrafish, a common model used in biology to study development but also human diseases. In this study, we have investigated the expression of S100 protein in the central nervous system of adult zebrafish using PCR, Western blot, and immunohistochemistry. The central nervous system of the adult zebrafish express S100 protein mRNA, and contain a protein of approximately 10 kDa identified as S100 protein. S100 protein immunoreactivity was detected widespread distributed in the central nervous system, labeling the cytoplasm of both neuronal and non-neuronal cells. In fact, S100 protein immunoreactivity was primarily found in glial and ependymal cells, whereas the only neurons displaying S100 immunoreactivity were the Purkinje's neurons of the cerebellar cortex and those forming the deep cerebellar nuclei. Outside the central nervous system, S100 protein immunoreactivity was observed in a subpopulation of sensory and sympathetic neurons, and it was absent from the enteric nervous system. The functional role of S100 protein in both neurons and non-neuronal cells of the zebrafish central nervous system remains to be elucidated, but present results might serve as baseline for future experimental studies using this teleost as a model.