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C Redies

Publications and source records attributed to C Redies.

At least 55 records · Page 3Linked to original sources
PubMed1993

Expression of N-cadherin mRNA during development of the mouse brain.

The expression of N-cadherin mRNA was mapped in the brain of mice between embryonic day 12 (E12) and the adult stage by in situ hybridization of digoxigenin-labeled riboprobe. Two phases of N-cadherin expression can be distinguished. During the first phase (about E12 to E16), expression is ubiquitous throughout the brain and most prominent in the proliferative neuroepithelium. During the second phase (about E16 to postnatal day 6), N-cadherin expression is restricted to particular nuclei or laminae that share common functional features and neuroanatomical connections. Several of the N-cadherin-positive structures receive direct afferents from retinal ganglion cells or from the superior colliculus. Others belong to the reticular system and to the limbic system of the brain. In neocortex, N-cadherin is expressed by deeper layer cells. In the adult brain, only low levels of N-cadherin expression remain in very few types of cells, for example in the Purkinje cells of the cerebellum. These results are similar to data from chicken brain and suggest that the generalized expression of N-cadherin during the early phase and the restriction expression of this molecule in particular functional systems during the later phase is, at least in part, phylogenetically conserved between chicken and mouse. Moreover, the results show that N-cadherin expression extends to phylogenetically newer structures, e.g., the mammalian neocortex.

Animals↗
PubMed1993

N- and R-cadherin expression in the optic nerve of the chicken embryo.

Cadherins are a family of molecules mediating Ca(2+)-dependent cell-cell adhesion in various tissues. N- and R-cadherin are expressed in the chick embryonic CNS and differ in their expression pattern during development. Here we focus on the differential expression of N- and R-cadherin in the early optic nerve. N-cadherin is expressed by the retinal neurites growing through the optic nerve. R-cadherin is expressed by the early optic nerve glia, which derives from the optic stalk neuroepithelium and corresponds to an immature form of the type-1 astrocyte described in rat optic nerve. The close contact between the plasma membranes of the retinal neurites and the optic nerve glia is believed to be important in guiding retinal axons through the optic nerve. Using neuroblastoma cell lines transfected with R-cadherin, we demonstrate that the N-cadherin-positive retinal axons can use R-cadherin as a substrate for axon elongation. These results suggest that the R-cadherin expressed by the early optic nerve glia might provide a molecular substrate for the growth of N-cadherin-positive retinal axons through the optic nerve.

Animals↗
PubMed1992

Restricted expression of N- and R-cadherin on neurites of the developing chicken CNS.

The expression of two cadherins, N- and R-cadherin, was mapped in the CNS of chicken embryos of 6-11 d incubation, focusing on the sensory and motor fiber systems. In the spinal cord, the laterally located fibers of the dorsal funiculus express N-cadherin while the medially located fibers do not. These two fiber systems have a different course within the CNS but associate to form the spinal dorsal roots. In the hindbrain, N-cadherin is expressed by the descending trigeminal (general somatic sensory) tract, which is contiguous with the N-cadherin-positive zone of the dorsal funiculus of the spinal cord. R-cadherin is not expressed by sensory fibers, but is expressed by the visceral motor system of the vagus and glossopharyngeal nerves, which are N-cadherin negative. The motor neurites expressing R-cadherin have a different course within the brain than the sensory neurites expressing N-cadherin, although they form the common sensory/motor roots of the vagus nerve at the surface of the brain. The possibility that N-cadherin provides a guidance cue for sensory axon migration within the CNS by a homophilic adhesion mechanism was investigated in vitro. Explants from sensory spinal ganglia expressing N-cadherin were placed on N-cadherin-transfected neuroblastoma cells, and axon outgrowth was visualized. Results showed that the sensory axons defasciculate and closely follow the cell-cell boundaries between transfected cells where high levels of N-cadherin are expressed. These results show that the two cadherins, like members of the immunoglobulin superfamily of molecules, are expressed in a topographically restricted fashion during chick brain development. They furthermore suggest that N-cadherin expression by neurites may play a role in guiding these neurites along CNS paths that express the same molecule.

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PubMed1991

Differentiation and heterogeneity in T-antigen immortalized precursor cell lines from mouse cerebellum.

Recently, various techniques have been developed to transfer oncogenes into brain cells in order to generate immortalized neural cell lines. It is of interest to establish how well such cell lines reflect their cellular origin. Here we report the characterization of sixteen cell lines from mouse cerebellum and, as a control, six cell lines from skin. Lines were established by immortalizing postnatal primary cell cultures with a retrovirus carrying a modified temperature-sensitive variant of SV40 large T antigen. The cell lines reflect many properties of the cell type from which they were derived. All of the sixteen cerebellar lines expressed one or more markers of the neural precursor cells, namely, nestin and epitopes for NG2 and A2B5. In contrast, none of the six skin lines expressed neural precursor markers. Both types of cell lines expressed vimentin and fibronectin. Differentiation occurred in some of the cerebellar lines and was enhanced in defined medium. A small percentage of cerebellar cells, usually less than 5%, was positive for a marker of differentiation, e.g., glial fibrillary acidic protein (GFAP), galactocerebroside (GalC), or L1. Expression of GFAP colocalized with that of nestin at varying levels of intensity, indicating a gradual replacement of nestin by GFAP in the cytoskeleton. Both the cells positive for precursor markers and those positive for differentiation markers tended to be located in clusters, suggesting that stochastic processes or cell-cell interactions are important for the determination of the fate of cells within a clonal cell line in vitro. The degree of differentiation seemed to correlate with a shift from serum-containing to defined medium, but not with a shift from the permissive to the nonpermissive temperature for T antigen expression. The immortalization approach described here thus allows the establishment of cell lines which are "captured" in the precursor state of the developing mouse neuroepithelium.

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PubMed1991

Differential expression of R- and N-cadherin in neural and mesodermal tissues during early chicken development.

R-cadherin is a newly identified member of the cadherin family of cell adhesion receptors. The expression of R-cadherin in early chicken embryos was studied using affinity-purified antibodies to this molecule, comparing it with that of N-cadherin. Immunoblot analysis of various organs of 10.5-day embryos showed that R-cadherin is most abundantly expressed in the retina and brain. Immunostaining of the cervical and thoracic regions of embryos revealed that R- and N-cadherin are expressed in all neural tissues. In the neural tube, R-cadherin appears at around stage 21, although N-cadherin expression begins at a much earlier stage. The distribution of R-cadherin in the neural tube differs from that of N-cadherin; for example, some regions of the tube express only R-cadherin, and other regions only N-cadherin. In the peripheral ganglia, these two cadherins are also expressed in different patterns which change during development. Some mesenchymal tissues including the notochord, the myotome, myotubes and perichondria also express these cadherins, again in different patterns. Thus, R- and N-cadherin are differentially expressed in all the tissues examined, and they may contribute to the spatial segregation of heterogeneous cells in a tissue.

Animals↗
PubMed1990

Functional organization in the ferret visual cortex: a double-label 2-deoxyglucose study.

The functional organization of visual cortical area 17/18 of the ferret was studied using a double-label 2-deoxyglucose (2-DG) method. Animals were stimulated sequentially with moving gratings of 2 different orientations. Elongated orientational patches running roughly at a right angle into the area 17/18 border were seen. The orientation maps were similar in areas 17 and 18. When animals were stimulated with 2 orthogonally oriented gratings, activation patterns were not fully complementary but did not overlap. A complex pattern of partial overlap was observed, with orientations differing by 45 degrees. More gradual and more abrupt changes alternated frequently on a scale smaller than the average periodicity in the patterns. A cortical patch representing a given orientation was regularly surrounded by both neighboring and orthogonal orientations. The direction of the orientational changes reversed frequently in the immediate vicinity of any orientational patch. Orientation maps were compared with complete maps of retino-cortical projections obtained by transneuronal labeling with intravitreally injected tritiated proline. Ocular dominance in the binocular segment of both areas 17 and 18 was found to be organized into elongated ipsilateral islands in an almost contiguous contralateral projection. In area 18, the patches were wider than in area 17, and the ipsilateral islands were smaller in the regions representing the upper and central visual field than in those representing the lower visual field.

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PubMed1989

Discontinuities along lines: psychophysics and neurophysiology.

If a segment of a line differs in luminance or color from the rest of the line, three illusory phenomena may be perceived: a reduction in contrast of the line segment relative to the background, subjective contours running perpendicularly to the ends of the line segment, and spread of color or brightness surrounding the line segment. All three phenomena cause a perceptual enhancement of the line discontinuity. Their psychophysical properties and possible neurophysiological correlates in the visual cortex are discussed. Contrast reduction has characteristics similar to end-stopped neuronal responses. Subjective contours elicit orientation-specific neuronal responses in areas 17 and 18. Neon color and brightness spreading may be mediated by "dot-responsive" cells in area 19.

Animals↗
PubMed1989

The deoxyglucose method in the ferret brain. I. Methodological considerations.

In the brain of the anesthetized ferret, the 2-deoxyglucose (2-DG) transfer rate constants required to determine cerebral glucose utilization by the deoxyglucose method were calculated from regional gray matter time-radioactivity curves measured for 180 min after tracer injection. Results suggest that loss of metabolized tracer from brain occurs at a rate of about 1%/min for the first 180 min after injection if the rate constant of the rate-limiting step for loss of metabolized tracer (k4*) represents a first-order kinetic process. A simulation experiment shows that, whether k4* is assumed to be 0 or 0.01 min-1, has a negligible influence on glucose utilization rates obtained in conventional 45 min autoradiographic experiments provided that the entire analysis, including lumped constant determination, is carried out in a consistent way. The 2-DG lumped constant for k4* = 0 is 0.54, and 0.68 for k4* = 0.01 min-1.

Animals↗
PubMed1989

The deoxyglucose method in the ferret brain. II. Glucose utilization images and normal values.

To measure cerebral glucose utilization with the autoradiographic deoxyglucose method, the tracer transfer rate constants and lumped constants must be known. 2-Deoxyglucose (2-DG) and fluorodeoxyglucose (FDG) constants were determined in 18 gray and white matter brain structures of the anesthetized ferret. The ferret is a domestic carnivore particularly suitable for deoxyglucose studies because of its small brain size and low body weight. The average gray matter rate constants for tracer transfer across the blood-brain barrier are similar for 2-DG and FDG in the ferret brain (K*1 = 0.21 ml/g/min and k*2 = 0.39 min-1). The rate constant for the rate-limiting step of tracer phosphorylation, k*3, is 1.6 times higher for FDG than for 2-DG (0.21 vs. 0.13 min-1). Loss of metabolized tracer is about 1-1.5%/min throughout the ferret brain for both tracers as estimated for a 180 min experimental period. Taking into account this loss, the lumped constant is 0.92 for FDG and 0.68 for 2-DG. Glucose utilization values in the brain of the anesthesized ferret range from 33 mumol/100 g/min in the corpus callosum to 104 mumol/100 g/min in the caudate nucleus. Representative glucose utilization images of coronal sections of the ferret brain are shown. Brain structures are identified on the same slices counterstained with thionin.

Animals↗
PubMed1989

Generalized decrease in brain glucose metabolism during fasting in humans studied by PET.

In prolonged fasting, the brain derives a large portion of its oxidative energy from the ketone bodies, beta-hydroxybutyrate and acetoacetate, thereby reducing whole body glucose consumption. Energy substrate utilization differs regionally in the brain of fasting rat, but comparable information has hitherto been unavailable in humans. We used positron emission tomography (PET) to study regional brain glucose and oxygen metabolism, blood flow, and blood volume in four obese subjects before and after a 3-wk total fast. Whole brain glucose utilization fell to 54% of control (postabsorptive) values (P less than 0.002). The whole brain rate constant for glucose tracer phosphorylation fell to 51% of control values (P less than 0.002). Both parameters decreased uniformly throughout the brain. The 2-fluoro-2-deoxy-D-glucose lumped constant decreased from a control value of 0.57 to 0.43 (P less than 0.01). Regional blood-brain barrier transfer coefficients for glucose tracer, regional oxygen utilization, blood flow, and blood volume were unchanged.

Adult↗
PubMed1989

Double-label and conventional deoxyglucose methods: a practical guide for the user.

The autoradiographic deoxyglucose method is widely used to map functional activity in mammalian brain. Whereas the method is simple to use, the underlying kinetic model is complex. This paper reviews the deoxyglucose kinetic model and the relevant implications for the user who does not have extensive knowledge of tracer kinetics. In generally understandable terms, single-label and double-label deoxyglucose approaches are discussed. Experimental procedures are described in detail. The calculations required for qualitative and quantitative experiments are explained. The deoxyglucose method is compared to other methods that map functional activity in mammalian brain.

Animals↗
PubMed1988

Influx of a choline analog to dog brain measured by positron emission tomography.

The influx of the 11C-labeled choline analog pyrrolidinocholine into tissue was measured in the brain of three dogs by positron emission tomography (PET). During the first 90 s after the intravenous bolus injection of the tracer, transfer of tracer from plasma to tissue was unidirectional. The influx constant for pyrrolidinocholine into intracranial tissue, Kin, was 0.017 ml/g/min (0.008 SD), and the initial volume of distribution, V0, was 0.08 ml/g (0.03 SD). The influx constant was at least five times larger than the value expected if simple diffusion were to account for tissue uptake. The method presented in this paper can be used to investigate the availability of plasma choline and its analogs to the living human brain and other tissue in degenerative diseases affecting the cholinergic system, and to provide in vivo information on a choline transport system.

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PubMed1987

In vivo measurement of [18F]fluorodeoxyglucose rate constants in rat brain by external coincidence counting.

The operational equation for the double-label deoxyglucose method described in the following paper requires the knowledge of the rate constants for transfer of fluorodeoxyglucose across the blood-brain barrier (K1* and K2*), and those for phosphorylation of fluorodeoxyglucose (K3*) and dephosphorylation of fluorodeoxyglucose-6-phosphate (k4*). These rate constants were determined in anesthetized rats by external coincidence counting. Radioactivity in parietal brain was measured for a 110 min experimental period after a bolus injection of 18F-labeled fluorodeoxyglucose. Apparent rate constants were obtained by fitting the resulting tissue radioactivity curves to the tissue radioactivity function of the deoxyglucose model modified to take into account the dephosphorylation of fluorodeoxyglucose-6-phosphate. The apparent fluorodeoxyglucose rate constants in rat brain are K1* = 0.195 ml g-1 min-1, k2* = 0.379 min-1, k3* = 0.088 min-1, and k4* = 0.009 min-1.

Animals↗
PubMed1987

Double-label autoradiographic deoxyglucose method for sequential measurement of regional cerebral glucose utilization.

A new double-label autoradiographic glucose analog method for the sequential measurement of altered regional cerebral metabolic rates for glucose in the same animal is presented. This method is based on the sequential injection of two boluses of glucose tracer labeled with two different isotopes (short-lived 18F and long-lived 3H, respectively). An operational equation is derived which allows the determination of glucose utilization for the time period before the injection of the second tracer; this equation corrects for accumulation and loss of the first tracer from the metabolic pool occurring after the injection of the second tracer. An error analysis of this operational equation is performed. The double-label deoxyglucose method is validated in the primary somatosensory ("barrel") cortex of the anesthetized rat. Two different rows of whiskers were stimulated sequentially in each rat; the two periods of stimulation were each preceded by an injection of glucose tracer. After decapitation, dried brain slices were first exposed, in direct contact, to standard X-ray film and then to uncoated, "tritium-sensitive" film. Results show that the double-label deoxyglucose method proposed in this paper allows the quantification and complete separation of glucose utilization patterns elicited by two different stimulations sequentially applied in the same animal. The double-label deoxyglucose is of potential usefulness in sensory physiology since it makes possible the separate mapping of regional cerebral glucose utilization patterns elicited by two sequentially applied sensory stimulations in the same animal. The method allows the quantification of a step-like change in regional cerebral glucose utilization in the same animal. It could be used to study the cerebral metabolic effects induced by neuropharmacological agents or surgical interventions applied during the experiment. Using each animal as its own control eliminates intersubject variability. Thus experimental cost and effort can be saved, and the reliability of the results obtained can be increased.

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