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Expression of cadherin-11 delineates boundaries, neuromeres, and nuclei in the developing mouse brain.

Cadherin-11 (cad11 or OB-cadherin) was previously identified as a mesenchymal cell-cell adhesion molecule. Here we studied the expression of cad11 transcripts in developing brains derived from E11.5 to E16.5 mouse embryos. In the brains at these stages, cad11 was expressed in various patterns, which could be grouped into three categories. First, cad11 expression occurred along boundaries between certain brain subdivisions, including those between the ventral and dorsal thalamus and between the mesencephalon and metencephalon. At these boundaries, cad11-positive cells were localized in a narrow, columnar compartment of the neuroepithelium. Second, cad11 expression delineated particular neuromeric compartments at the ventricular zone of the neuroepithelium. A typical example of this pattern was observed in the hypothalamus. Third, cad11 was expressed in differentiating or differentiated brain nuclei. The former included the thalamus, epithalamus, and pretectum; the latter included the mammillary, red, trigeminal motor, facial motor, prepositus hypoglossal, and inferior olive nuclei, as well as the substantia nigra. Furthermore, developing nuclei and the superficial zone of the cerebellum expressed cad11, and the cortical plate of the developing cerebrum also did so. We compared the expression pattern of cad11 with that of R-cadherin in the diencephalon and found that each cadherin delineated a unique set of diencephalic subdivisions. These findings suggest that cad11-mediated specific cell-cell adhesion plays roles in segmentation or compartmentalization of the developing brain in various ways. We also discussed the possibility that cad11 might be involved in neuronal connections between specific nuclei.

Animals↗

Marsupial retinocollicular system shows differential expression of messenger RNA encoding EphA receptors and their ligands during development.

The protracted development of the wallaby (Macropus eugenii) has allowed study of messenger RNAs encoding Eph receptors EphA3 and EphA7 and ligands ephrin-A2 and -A5 in the retina and superior colliculus at intervals throughout the development of the retinocollicular projection: from birth, before retinal innervation, to postnatal day 95, when the projection is mature. Reverse transcription-polymerase chain reaction showed messenger RNAs for both receptors and ligands were expressed at all ages. EphA7 was expressed more highly in the rostral superior colliculus. Ephrin-A2 and -A5 were expressed more highly in the caudal colliculus. EphA3 was expressed in a complementary manner, more highly in temporal than in nasal retina. There are higher levels of expression of the ligands when the projection is only coarsely topographically organised. This suggests a role for them and their receptor EphA3 in this stage, by repulsive interactions which restrict temporal axons to rostral superior colliculus. This is the first account in a marsupial mammal of the appearance of this molecular family, substantiating its ubiquitous role in topographically organised neuronal connections. Nevertheless, expression is not the same as in the mouse, suggesting differences in the details of topographic coding between species.

Animals↗

Walking and fatigue in multiple sclerosis: the role of the corticospinal system.

Transcranial magnetic stimulation was used to study motor evoked potentials (MEPs) of leg muscles in controls and patients with multiple sclerosis (MS) before and after walking. In controls, MEP areas were significantly reduced after walking. A similar or greater reduction was seen in most patients, although there was a wide range of values. The M waves were unchanged. We conclude that walking induces functional changes of the corticospinal system and/or connected neurons contributing to central fatigue, especially in patients with MS.

Adult↗

Developmentally regulated expression of a cell surface protein, neuropilin, in the mouse nervous system.

Neuropilin (previously A5) is a cell surface glycoprotein that was originally identified in Xenopus tadpole nervous tissues. In Xenopus, neuropilin is expressed on both the presynaptic and postsynaptic elements in the visual and general somatic sensory systems, suggesting a role in neuronal cell recognition. In this study, we identified a mouse homologue of neuropilin and examined its expression in developing mouse nervous tissues. cDNA cloning and sequencing revealed that the primary structure of the mouse neuropilin was highly similar to that of Xenopus and that the extracellular segment of the molecule possessed several motifs that were expected to be involved in cell-cell interaction. Immunohistochemistry and in situ hybridization analyses in mice indicated that the expression of neuropilin was restricted to particular neuron circuits. Neuropilin protein was localized on axons but not on the somata of neurons. The expression of neuropilin persisted through the time when axons were actively growing to form neuronal connections. These observations suggest that neuropilin is involved in growth, fasciculation, and targeting for a particular groups of axons.

Amino Acid Sequence↗

Addiction molecular genetics: 639,401 SNP whole genome association identifies many "cell adhesion" genes.

Addictions are substantially heritable complex disorders. We now report whole genome association studies that identify 89 genes likely to contain variants that contribute to addiction vulnerability, using previously- and newly-validated microarray based pooling assays. Each gene contains clustered single nucleotide polymorphisms (SNPs) that display significant allele frequency differences between abusers and controls in each of the two samples studied with 639,401 SNP arrays and confirmatory SNPs from each of two other abuser/control samples. These genes are implicated in interesting functions, including "cell adhesion" processes that help to establish and maintain neuronal connections of special relevance to addiction's memory-like features.

Adult↗

In situ injection of kainic acid: a new method for selectively lesioning neural cell bodies while sparing axons of passage.

The morphologic sequelae following stereotaxic injection into the rat striatum of kainic acid, a conformationally restricted analogue of glutamate, were examined by means of bright field, histofluorescence and electron microscopic techniques. The neuropathologic response to kainate injection occurs in two distinct phases. First, the intrinsic neurons of the striatum undergo a rapid degeneration during the first 48 hours after injection; this is characterized by the sequential loss of cytoplasmic Nissl-substance (chromatolysis), shrinkage of the perikarya, clumping of the nuclear chromatin and finally disruption of the nuclear membrane. Between one and three weeks after injection, a marked proliferation of astrocytes in the gray matter formerly populated by neurons characterizes the second phase. The region of several neuronal loss in the kainate injected striatum is approximately spherical in shape, and its extent is non-linearly related to the amount of kainate injected. The neuropil of the injected striatum is markedly disrupted due to the death of intrinsic neurons and loss of their processes. Yet, histofluorescence microscopy demonstrates that the dopaminergic axons projecting from the substantia nigra do not degenerate in the kainate injected striatum; electron microscope studies indicate that corticofugal fibers traversing the striatum are also unaffected by kainate. Many presynaptic boutons, presumably of extrinsic origin, are intact up to ten days after injection; osmophilic vestiges of postsynaptic elements remain adherent to these boutons. Numerous phagocytic astrocytes are observed throughout the lesioned area. These morphologic studies provide (direct) evidence that in situ injection of kainic acid in brain causes a selective degeneration of neurons with cell bodies in the area of the injection but spares axons that arise from perikarya outside the region but pass through or terminate in the injected area. Thus, in situ injection of kainic acid is a new technique for making selective brain lesions that will be useful for examining neuronal connectivity.

Animals↗

Morphological study of noradrenaline innervation in the caudal raphe nuclei with special reference to fine structure.

Previous histofluorescence studies have demonstrated that the caudal raphe nuclei (CRa) of the rat, particularly the ventral portion (VCRa), contains a very high density of noradrenaline (NA) terminals and in the present study we attempted to elucidate the origins and free structure of the NA terminals in this region. The majority of NA terminals found in the VCRa originated from A1 and A3 NA neurons and disclosed that a small number of very strongly fluorescent fibers located along the blood vessels arise from the superior cervical ganglion (SGC). Electron microscopic analysis after potassium permanganate fixation demonstrated that the NA terminals originated from A1 and A3 NA neurons are connected with neuronal elements, while NA fibers from the SGC were identified among the vascular elements. Axodendritic contacts were found to be predominant among the synapselike contacts of NA terminals in this area.

Animals↗

Postnatal changes in the number of astrocytes, oligodendrocytes, and microglia in the visual cortex (area 17) of the macaque monkey: a stereological analysis in normal and monocularly deprived animals.

The numerical densities (Nv) of astrocytes, oligodendrocytes, and microglia were measured in individual laminae of the striate cortex of macaque monkeys ranging in age from newborn to adult. Using measurements of cortical thickness and surface area, the total number of cells in the striate cortex of one hemisphere was derived for each glial cell type. Normal monkeys were compared at 3 months and 6 months of age to animals reared from birth with a monocular eyelid suture. No significant differences were observed between normal and monocularly deprived monkeys. The combined data from these groups, however, demonstrated several significant developmental changes. The Nv of astrocytes decreased from birth to 6 months of age and subsequently increased in the adult. The greatest changes were seen in the more superficial laminae. These changes, however, were only a response to a substantial overshoot in cortical volume at six months: the total number of astrocytes in the striate cortex did not change. There was a tenfold increase in both the Nv and the total number of oligodendrocytes from birth to maturity with a corresponding increase in the density of myelinated axons. The greatest changes were observed in the deeper laminae. The total number of microglia remained relatively constant from birth to 6 months of age. There was a 55% reduction in the number of microglia in the adult, although statistical analysis indicated that this decrease was only of borderline significance. The possible relationships between these postnatal changes in glial cell numbers and the development of neuronal connectivity are discussed.

Aging↗

Neurotensin in projection neurons of the striatum and nucleus accumbens, with reference to coexistence with enkephalin and GABA: an immunohistochemical study in the cat.

Neurotensin-like immunoreactivity (NT-LI) was demonstrated in projection neurons of the striatum and nucleus accumbens in the cat by combining immunohistochemistry and the fluorescent retrograde neuronal labeling method. In colchicine-treated cats, many neurons with NT-LI were found in the caudate nucleus, nucleus accumbens, and putamen. Most of these neurons were medium-sized neurons with spiny dendrites. NT-LI of neuronal elements in the caudate nucleus and nucleus accumbens formed dense aggregates with irregular figures, which appeared to correspond to the striosomes of Graybiel et al. (Proc. Natl. Acad. Sci. USA 75:5723-5726, '78; Exp. Brain Res. 34:189-195, '79; Neuroscience 6:377-397, '81). Fibers with NT-LI were distributed massively to the globus pallidus and ventral midbrain regions, but not to the entopeduncular nucleus. In the ventral midbrain regions, many fine varicose fibers with NT-LI were distributed to the pars compacta and pars lateralis of the substantia nigra, ventral tegmental area, and retrorubral area. In the pars reticulata of the substantia nigra, however, fibers with NT-LI were rather sparse. Examination of consecutive sections immunostained for NT, enkephalin (Enk), GABA, and substance P (SP) revealed that 50% of neurons with NT-LI in the caudate nucleus and nucleus accumbens exhibited Enk-LI, 15% showed GABA-LI, and 5% manifested both Enk-LI and GABA-LI; no NT-positive neurons in the striatum and nucleus accumbens showed SP-LI. No morphological differences were found between NT-positive neurons with Enk-LI and/or GABA-LI and those without Enk-LI and GABA-LI. Most neurons with NT-LI in the striatum and nucleus accumbens were retrogradely labeled with True Blue injected into the globus pallidus, pars compacta and pars lateralis of the substantia nigra, and ventral tegmental area. After hemitransection severing neuronal connections between the ventral midbrain regions and the forebrain structures, fibers with NT-LI and those with Enk-LI in the ventral midbrain regions were markedly reduced in number.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Organization of the projection from the superficial to the deep layers of the hamster's superior colliculus as demonstrated by the anterograde transport of Phaseolus vulgaris leucoagglutinin.

Anterograde tracing with Phaseolus vulgaris leucoagglutinin (PHA-L) was employed to describe the projection from the superficial to the deep layers of the hamster's superior colliculus (SC). Deposits of PHA-L in the stratum griseum superficiale (SGS) resulted in labelled terminal swellings in the stratum opticum and all of the deep laminae (the stratum griseum intermediate [SGI], stratum albumin intermedium [SAI], stratum griseum profundum [SGP], and stratum albumin profundum [SAP]). Labelled terminals were also visible in the periaqueductal gray (PAG). Reconstructions of individual axons showed that many collateral in the deep laminae arose from axons that projected to targets outside the colliculus. The projection from the superficial to the deep laminae had a loose topographic organization, and the trajectories of interlaminar axons were generally deflected laterally from "projection" lines that were orthogonal to the SC surface. Physiological recording and receptive field mapping were used to determine actual projection lines, which connect neurons in the superficial and deep layers that have receptive fields with the same elevation. These projection lines closely matched the trajectory of the pathway from the superficial to the deep laminae.

Animals↗

Deprived somatosensory-motor experience in stumptailed monkey neocortex: dendritic spine density and dendritic branching of layer IIIB pyramidal cells.

Infant macaque monkeys (Macaca arctoides) were individually raised to age 6 months in large clear cubes built into one wall of a control colony that allowed them visual access to it but not tactile contact. The two deprivation conditions (Cond 2 and Cond 3) were equal both in physical size and with respect to partial social isolation. They differed in the degree of somatosensory-motor opportunity available during development in that the Cond 2 chamber was empty, whereas Cond 3 contained ladders, a trapeze, and play objects. Four monkeys from each of these conditions were compared with four colony-reared (Cond CR) monkeys. Neuroanatomical changes were evaluated by using light microscopy in Golgi-Cox-stained neocortex. Dendritic spines on the apical shafts of layer IIIB pyramidal cells were counted in primary motor (MI), somatosensory (SI), and visual (area 17, V1) cortical regions. Layer IIIB pyramidal neurons with somas of medium size were selected from each cortical region and the density of apical dendritic spines determined. The basilar dendritic branches of these same neurons were traced, and the dendritic branching complexity was assessed in order to compare the sensitivity of the dendritic spine and branching measures consequent to deprived rearing. The number of apical dendritic spines was significantly reduced in Cond 2 when compared with either Cond 3 or Cond CR (which did not differ from each other). This occurred in both MI and SI cortex, but not in the visual cortex, the region used as a control for a global brain effect. Branching complexity measured on the same pyramidal neurons was reduced only in MI cortex of Cond 2. These results show spine density, a more direct measure of neuronal connectivity, to be the more sensitive measure of early environmental deprivation. Also, the enriched environment provided by Cond 3 relative to Cond 2 offset the effect of partial social isolation such that both morphometric measures had values comparable to Cond CR monkeys.

Animals↗

Immunohistochemical distribution and partial characterization of FLFQPQRFamidelike peptides in the central nervous system of rats.

Two peptides that are structurally related to the molluscan tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRF-NH2) were recently isolated from bovine brain extract (Yang et al.: Proc. Natl. Acad. Sci. USA 82:7757-7761, '85). These peptides have an attenuating effect on morphine-induced analgesia when injected intracerebroventricularly in rats. Antisera against the two peptides--an octapeptide, Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2 (F8F-NH2), and an octadecapeptide, Ala-Gly-Glu-Gly-Leu-Ser-Ser-Pro-Phe-Trp-Ser-Leu-Ala-Ala-Pro-Gln-Arg-Phe- NH2 (A18F-NH2)--were raised in rabbits and characterized with standard radioimmunoassay and immunohistochemical blocking controls. This study was aimed at localizing neurons in the rat brain that contain immunoreactivity for these peptides. Cryostat sections of normal and colchicine-treated Sprague-Dawley rat brains were incubated with the specific antisera and the immunoreactivity was visualized by the PAP or the FITC method. Immunoreactive neurons were detected in the hypothalamus and nucleus of the solitary tract. Relatively dense networks of fibers and terminals were observed in the lateral parabrachial nucleus and in the nucleus of the solitary tract. Fibers and terminals were also seen in the lateral septum, amygdala, hypothalamus, neurohypophysis, thalamus, periaqueductal gray, and several medullary nuclei. In these areas the immunoreactivity was abolished when the antisera were incubated with the corresponding synthetic peptides F8F-NH2 and A18F-NH2. Preincubation of the antisera with neuropeptide Y (NPY) did not affect the staining. The results suggest that there are neurons in the rat brain that contain F8F-NH2- and A18F-NH2-like peptides. The neuronal system described here may have a role in the regulation of autonomic functions, such as hypothalamic control and respiratory functions. The neuronal connections of the cells containing F8F-NH2- and A18F-NH2-like peptides remain to be studied.

Amino Acid Sequence↗

Genetic control of retinal projections in inbred strains of albino mice.

Mutations in the tyrosinase gene are often associated with a misrouting of retinal ganglion cell axons at the optic chiasm. In albinos, tyrosinase activity is lost and some ganglion cell axons that would normally project into the ipsilateral optic tract instead cross midline and project into the contralateral tract. The developmental mechanisms that cause this modification in neuronal connectivity are unknown. In this study, we screened six diverse strains of albino mice (strains 129, A, AKR, BALB/c, C57BL/6-c/c, and CD-1) to discover genetically determined variations and possible gene loci that might affect the severity of the albino decussation abnormality. Ganglion cells were retrogradely labeled with horseradish peroxidase, and the ipsilaterally and contralaterally projecting cells were counted. The average number of ipsilaterally projecting ganglion cells in the six albino strains varies from 1,000 to 1,300. Despite this variation, 1.8-1.9% of the total population projects ipsilaterally in each strain. In comparison, 2.8% project ipsilaterally in the pigmented strain, C57BL/6(-)+/+. However, the percentage of displaced, ipsilaterally projecting cells varies substantially among albino strains--from a low of 4% in strain CD-1 to a high of nearly 10% in C57BL/6-c/c. We conclude that even with large differences in genetic background and in absolute numbers of ganglion cells, there is no appreciable variation in the magnitude of decussation error among albino mice. The consistent effect of null alleles at tyrosinase suggests a comparably tight linkage between the biochemical activity of this enzyme and the mechanisms that control decussation phenotype.

Alleles↗

Semaphorin/neuropilin signaling influences the positioning of migratory neural crest cells within the hindbrain region of the chick.

Within the hindbrain region, neural crest cell migration is organized into three streams that follow the segmentation of the neuroepithelium into distinct rhombomeric compartments. Although the streaming of neural crest cells is known to involve signals derived from the neuroepithelium, the molecular properties underlying this process are poorly understood. Here, we have mapped the expression of the signaling component of two secreted class III Semaphorins, Semaphorin (Sema) 3A and Sema 3F, at time points that correspond to neural crest cell migration within the hindbrain region of the chick. Both Semaphorins are expressed within rhombomeres at levels adjacent to crest-free mesenchyme and expression of the receptor components essential for Semaphorin activity by neural crest cells suggests a function in restricting neural crest cell migration. By using bead implantation and electroporation in ovo, we define a role for both Semaphorins in the maintenance of neural crest cell streams in proximity to the neural tube. Attenuation of Semaphorin signaling by expression of soluble Neuropilin-Fc resulted in neural crest cells invading adjacent mesenchymal territories that are normally crest-free. The loss or misguidance of specific neural crest cell populations after changes in Semaphorin signaling also affects the integration of the cranial sensory ganglia. Thus, Sema 3A and 3F, expressed and secreted by the hindbrain neuroepithelium contributes to the appropriate positioning of neural crest cells in proximity to the neural tube, a process crucial for the subsequent establishment of neuronal connectivity within the hindbrain region.

Animals↗

Maturation of the GFAP-immunopositive glial network is delayed in the olfactory bulbs of rats subjected to early, low-dose X-irradiation.

Young rats were subjected to low-level, repeated doses of x-irradiation from a Co60 source and examined for the presence and arrangement of their GFAP-immunopositive system at the light and electron microscopic levels. In 2-week-old control rats, labelled cells were already distributed homogeneously throughout the bulb, while in the experimental animals, a (newborn- and early postnatal-type) radial arrangement of immunoreactive fibres continued to persist even until the end of the fourth postnatal week. This delay in the maturation of the GFAP-immunopositive glial system may contribute to the hindered development of the bulbar neurones, neuronal connections, and field potentials, as found in earlier studies.

Animals↗

Macrophages: important accessory cells for reproductive function.

Macrophages are found throughout reproductive tissues. To determine their role(s), we have studied mice homozygous for a null mutation (Csfm(op)) in the gene encoding the major macrophage growth factor, colony-stimulating factor-1 (CSF-1). Both male and female Csfm(op)/Csfm(op) mice have fertility defects. Males have low sperm number and libido as a consequence of dramatically reduced circulating testosterone. Females have extended estrous cycles and poor ovulation rates. CSF-1 is the principal growth factor regulating macrophage populations in the testis, male accessory glands, ovary, and uterus. However, analyses of CSF-1 nullizygous mice suggest that the primary reproductive defect is in the development of feedback regulation of the hypothalamic-pituitary axis. Although not correlating with deficiencies of microglia populations, electrophysiological investigations indicate an impairment of neuronal responses. This suggests that microglia, under the influence of CSF-1, act to organize neuronal connectivity during development and that the absence of this function results in a perturbation of the hypothalamic-pituitary-gonadal axis. Macrophages also appear to have functions in the differentiated tissues of the reproductive system, including having a positive influence on steroidogenic cells. These data suggest that macrophages, through their trophic functions, can be considered as essential accessory cells for normal reproductive functioning.

Animals↗

Expression of brain-derived neurotrophic factor mRNA in rat hippocampus after treatment with antipsychotic drugs.

Typical and atypical antipsychotic drugs, though both effective, act on different neurotransmitter receptors and are dissimilar in some clinical effects and side effects. The typical antipsychotic drug haloperidol has been shown to cause a decrease in the expression of brain-derived neurotrophic factor (BDNF), which plays an important role in neuronal cell survival, differentiation, and neuronal connectivity. However, it is still unknown whether atypical antipsychotic drugs similarly regulate BDNF expression. We examined the effects of chronic (28 days) administration of typical and atypical antipsychotic drugs on BDNF mRNA expression in the rat hippocampus using in situ hybridization. Quantitative analysis revealed that the typical antipsychotic drug haloperidol (1 mg/kg) down-regulated BDNF mRNA expression in both CA1 (P < 0.05) and dentate gyrus (P < 0.01) regions compared with vehicle control. In contrast, the atypical antipsychotic agents clozapine (10 mg/kg) and olanzapine (2.7 mg/kg) up-regulated BDNF mRNA expression in CA1, CA3, and dentate gyrus regions of the rat hippocampus compared with their respective controls (P < 0.01). These findings demonstrate that the typical and atypical antipsychotic drugs differentially regulate BDNF mRNA expression in rat hippocampus.

Analysis of Variance↗

Allopregnanolone treatment, both as a single injection or repetitively, delays demyelination and enhances survival of Niemann-Pick C mice.

Niemann-Pick C disease (NPC) is an irreversible neurodegenerative disorder without current treatment. It is thought to result from deficient intracellular cholesterol and/or ganglioside trafficking. We have investigated the effects of allopregnanolone treatments on survival, weight loss, motor function, magnetic resonance imaging (MRI), and neuropathology in the mouse model of NPC (Npc1(-/-) mice). We confirmed previous results showing that a single injection of 250 microg of allopregnanolone on postnatal day 7 significantly extended the life span of Npc1(-/-) mice. This caused a marked difference in the weight curves of the treated mice but no statistical difference in the Rota-Rod performance. T2-weighted MRI and diffusion tensor imaging (DTI) of treated mice showed values of signal intensity and fractional anisotropy closer to those of wild-type mice than those of untreated Npc1(-/-) mice. Neuropathology showed that day-7 treatment markedly suppressed astrocyte reaction and significantly reduced microglial activation. Furthermore, the steroid treatment also increased myelination in brains of Npc1(-/-) mice. Similar effects of allopregnanolone treatment were observed in Npc1(-/-), mdr1a(-/-) double-mutant mice, which have a deficient blood-brain barrier, resulting in increased steroid uptake. The effects on survival and weight loss of a single injection on day 7 followed by injections every 2 weeks were also evaluated in Npc1(-/-) mice, and the beneficial effects were found to be greater than with the single injection at day 7. We conclude that allopregnanolone treatment significantly ameliorates several symptoms of NPC in Npc1(-/-) mice, presumably by effects on myelination or neuronal connectivity.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗