PubMed Health⌕ Search

Biomedical subjects

J Storm-Mathisen

Publications and source records attributed to J Storm-Mathisen.

At least 55 records · Page 3Linked to original sources
PubMed1987

The early development of neurons with GABA immunoreactivity in the CNS of Xenopus laevis embryos.

We have used an antibody to glutaraldehyde fixation complexes of gamma-amino butyric acid (GABA) to stain the developing central nervous system of Xenopus laevis embryos. Neuronal somata, growth cones, axons, and dendrites were found with GABA-like immunoreactivity. Transmission electron microscope (TEM) observations were made of axons and synapses. By observation of the earliest stages of differentiation of neurons, seven classes of putative GABAergic interneurons were discerned. 1) Ascending neurons are first stained in the hindbrain at stage 26 and later extend caudally in the spinal cord. They have ascending ipsilateral axons. 2) Midhindbrain reticulospinal neurons are first stained at stage 25 and develop as a compact group with descending ipsilateral and contralateral axons. 3) Vestibular complex commissural neurons are first stained at stage 29/30 in a dorsal position near the entry of the seventh and eighth cranial nerves. They have ventral commissural axons that descend contralaterally and their somata form a compact mass. 4) Rostral hindbrain commissural neurons are first stained at stage 33/34 just rostral to the entry of the trigeminal nerve. They each have a decussating projection. 5) Rostral midbrain neurons are first stained in the midbrain at stage 29/30 and are later associated with prominent dorsal and ventral commissures. 6) Optic tract and 7) rostral forebrain neurons are found in the forebrain associated with strongly stained axon tracts. The direction of axonal growth from its earliest stages was distinct for each class of hindbrain and spinal cord neuron.

Animals↗
PubMed1987

Postnatal development of neurons containing both catecholaminergic and GABAergic traits in the rat main olfactory bulb.

Postnatal development of catecholaminergic and gamma-aminobutyric acid (GABA)-ergic neurons in the periglomerular region of the rat main olfactory bulb was studied immunohistochemically using antisera against tyrosine hydroxylase (TH), glutamic acid decarboxylase and GABA. TH-like immunoreactive neurons almost always contained GABA-like immunoreactivity in the first postnatal week, but about 10% of them did not contain GABA-like immunoreactivity in older animals.

Aging↗
PubMed1987

Immunocytochemical localization of GABA in cat myenteric plexus.

Transverse sections through various levels of the feline gastrointestinal tract (antrum pyloricum, duodenum, jejunum, ileum and colon) were obtained from animals perfused with a mixture of glutaraldehyde and paraformaldehyde, and incubated with an antiserum specific for glutaraldehyde-fixed GABA. This antiserum has previously been characterized and shown to react selectively with presumed GABAergic neurons in the CNS. At all levels of the gastrointestinal tract, staining was observed in varicose fibers in the myenteric plexus and the circular muscle layer. Staining was abolished by glutaraldehyde complexes of GABA previously added to the antiserum, but not by similar complexes of related amino acids. The findings strongly suggest that GABA is concentrated in a subpopulation of gastrointestinal nerve fibers and add to previous evidence favoring a transmitter role for GABA in the enteric nervous system.

Animals↗
PubMed1987

Immunocytochemical demonstration of taurine.

Amino acid immunocytochemistry represents a new and powerful tool in neuroscience. Antisera are now available to a large number of amino acids, including those that are thought to serve a transmitter role. The antiserum described here against conjugated taurine seems to be of good specificity and can be used to demonstrate taurine in the CNS as well as in other organs; in normal tissue and in experimental and pathological conditions.

Animals↗
PubMed1987

Catecholaminergic neurons containing GABA-like and/or glutamic acid decarboxylase-like immunoreactivities in various brain regions of the rat.

The coexistence of immunoreactivities for tyrosine hydroxylase (TH) and glutamic acid decarboxylase (GAD) and/or gamma-aminobutyric acid (GABA) was revealed in various brain regions in colchicine-injected and untreated rats, using the peroxidase-antiperoxidase method. Consecutive 40 micron thick Vibratome sections were incubated in different antisera and those cells which were bisected by the plane of sectioning so as to be included at the paired surfaces of two adjacent sections were identified. The coexistence of the immunoreactivities for TH and GAD or GABA in the same cell could thus be determined by observing the immunoreactivity of the two halves of the cell incubated in two different antisera. In the olfactory bulb, retina, diencephalon, mesencephalic central grey and cerebral cortex, many TH-like immunoreactive neurons also showed GAD-like or GABA-like immunoreactivity, whereas in the substantia nigra, ventral tegmental area and locus ceruleus none of TH-like immunoreactive neurons showed either GAD-like or GABA-like immunoreactivity. In the olfactory bulb, retina and cerebral cortex, the majority of the TH-like immunoreactive neurons were also GAD-like or GABA-like immunoreactive. In the diencephalon of colchicine-injected rats, at least one-third of the TH-like immunoreactive neurons were GAD-like immunoreactive. Using serial 0.5 micron thick plastic-embedded sections, it was shown that immunoreactivities for three antigens, GAD, GABA and TH could occur in the same neurons in the olfactory bulb. These observations indicate the possible coexistence of two classical transmitters. GABA and catecholamine, in various brain regions of the rat.

Animals↗
PubMed1987

Glycine-like immunoreactivity in the cerebellum of rat and Senegalese baboon, Papio papio: a comparison with the distribution of GABA-like immunoreactivity and with [3H]glycine and [3H]GABA uptake.

An antiserum against conjugated glycine was characterized and applied to cerebellar sections of rats and baboons that had been perfusion-fixed with glutaraldehyde. After immunosorbent purification the serum reacted with brain protein-glutaraldehyde-glycine conjugates, but did not stain similar test conjugates prepared from other amino acids, including GABA and beta-alanine. In the rat cerebellum the glycine antiserum selectively labelled a subpopulation of Golgi neurons. Adjacent Vibratome sections treated with an antiserum against conjugated GABA revealed an about equally large subpopulation of immunopositive Golgi cells. A proportion of the Golgi cells that were cleaved by the plane of section contained both immunoreactivities. Additional evidence for a colocalization of glycine and GABA was obtained by postembedding staining of alternate semithin sections with the GABA antiserum and glycine antiserum, respectively. The ability of the antisera to distinguish between fixed glycine and GABA was corroborated by preincubation of the antisera with glutaraldehyde-amino acid fixation complexes: glycine complexes abolished staining with the glycine antiserum but had no effect on the GABA antiserum. The opposite effects were obtained with the GABA complexes. Matching the distributions of the respective immunoreactivities, [3H]glycine uptake was restricted to glomerulus-like structures in the granule cell layer whereas [3H]GABA uptake also occurred in punctate and fibrous profiles in the molecular layer. The baboon showed a distribution of glycine-like immunoreactivity similar to that in the rat, except that a few immunopositive neurons occurred in the molecular layer. The latter neurons were interpreted as outlying Golgi neurons; however, the possibility that they represent a subpopulation of basket cells could not be excluded. The Purkinje cells were negative in both species. Glial cells were weakly stained with the glycine antiserum but were strongly immunopositive after incubation with an antiserum raised against conjugates of the structurally similar amino acid beta-alanine. The present data suggest that glycine and GABA occur in about equally large subpopulations of Golgi neurons. A subpopulation of the Golgi neurons appears to contain both glycine and GABA.

Animals↗
PubMed1987

Gamma-aminobutyrate-like immunoreactivity in the thalamus of the cat.

Serial sections of the cat's thalamus were incubated with a purified antiserum raised against gamma-aminobutyric acid conjugated to bovine serum albumin by distilled glutaraldehyde. This serum has been extensively characterized and appears to react selectively with fixed gamma-aminobutyric acid in brain tissue treated with glutaraldehyde. Adjoining sections were stained with thionin and served as invaluable guides for a correct evaluation of the immunolabelling pattern. In the neuropil the intensity of the immunostaining varies considerably between thalamic nuclei and even between nuclear subdivisions. The neuropil staining appears particularly dense in the nuclei parataenialis, periventricularis, centralis medialis, reuniens, rhomboideus, habenularis lateralis, centrum medianum, parafascicularis, subparafascicularis, submedius, dorsal and ventral parts of the lateral geniculate body, the dorsal part of the medial geniculate body, the posterior complex, suprageniculate nucleus, pulvinar and parts of the lateral posterior nucleus. The pulvinar/lateralis posterior complex shows a particularly well-differentiated staining pattern which closely matches Updyke's [Updyke (1983) J. comp. Neurol. 219, 143-181] parcellation of this region. In several thalamic nuclei or subareas--and notably in those relay nuclei which are known to project upon non-primary sensory cortical areas--the immunostained neuropil is characterized by many puncta encircling an unstained profile. With few exceptions all thalamic nuclei displayed immunoreactive nerve cell bodies. Several examples were found of a mismatch between the number of such cells and the staining intensity of the neuropil. Thus the nuclei periventricularis, parafascicularis, subparafascicularis, parataenialis, limitans and centrum medianum although being very rich in neuropil staining have practically no immunostained perikarya. Rough estimates were made of the size and the proportion of gamma-aminobutyric acid labelled neurons in all major--and some minor--thalamic nuclei and their subdivisions. In some thalamic nuclei, notably the nuclei reticularis, anterior dorsalis, lateralis dorsalis, centralis lateralis, ventralis posterior and the dorsal lateral geniculate body, the population of immunoreactive neurons is distinctly heterogeneous with regard to soma size. The findings are discussed with regard to previous immunocytochemical studies of the distribution of gamma-aminobutyric acid and its synthesizing enzyme in the thalamus. Particular emphasis is put on the great species differences which appear to exist in this respect.

Animals↗
PubMed1987

Redistribution of transmitter amino acids in rat hippocampus and cerebellum during seizures induced by L-allylglycine and bicuculline: an immunocytochemical study with antisera against conjugated GABA, glutamate and aspartate.

The effects of the convulsants L-allylglycine and bicuculline on the distribution of gamma-amino-butyric acid (GABA), glutamate and aspartate in rat brains were assessed immunocytochemically, using antisera raised against glutaraldehyde-protein conjugates of the respective amino acids. In accord with previous biochemical studies of GABA content, L-allylglycine treatment was followed by a decreased immunoreactivity for GABA in the hippocampus and cerebellum, whereas treatment with bicuculline led to an increased immunoreactivity in the hippocampus, but not in the cerebellum. Different cells and zones were affected differentially. With both convulsants the hippocampus showed the most pronounced changes in the neuropil of the pyramidal and granular cell layers. L-Allylglycine treatment led to a substantial decrease in the concentration of detectable GABA-immunoreactive bouton-like dots in the stratum oriens, radiatum and lacunosum-moleculare and in the deep hilar region, but did not produce statistically significant changes in this parameter in the outer and intermediate zones of the dentate molecular layer. In the cerebellum, the decrease in GABA immunoreactivity after L-allylglycine treatment was less in the basket cell terminals than in other GABA-containing elements. Neither convulsant altered the average staining intensity for aspartate or glutamate in the two regions studied, but L-allylglycine reduced the level of aspartate-like immunoreactivity in hippocampal hilar cells. All the changes described were evident after 20 min of seizure activity and were qualitatively similar after 60 min of seizure (animals paralysed and ventilated). Our results indicate that L-allylglycine or bicuculline given intravenously exerts specific effects on cerebral amino acid metabolism. The nature and magnitude of these effects show inter-regional variations and also differ among cellular compartments within each region. Amino acid immunocytochemistry may prove to be a valuable tool for the investigation of metabolic changes associated with epileptic seizures and should be particularly useful in regions showing heterogeneous changes that would tend to cancel each other in biochemical analyses.

Allylglycine↗
PubMed1986

Implantation of D-[3H]aspartate loaded gel particles permits restricted uptake sites for transmitter-selective axonal transport.

Intracerebral pressure injections of small molecular tracers such as D-[3H]aspartate are usually followed by considerable spread which makes it difficult to assess the effective uptake area and precludes analysis of short axonal connections. As an attempt to circumvent these problems, D-[3H]aspartate was adsorbed onto gel beads that were subsequently packed into glass capillaries and implanted in the brain. Model experiments suggested that the loaded beads gradually release the tracer to the nerve tissue. The implantations resulted in small and sharply circumscribed tracer deposits which enabled us to study long axonal projections, as well as short intrahippocampal and intraamygdaloid pathways that are not easily resolved after pressure injections.

Amygdala↗
PubMed1986

Quantification of immunogold labelling reveals enrichment of glutamate in mossy and parallel fibre terminals in cat cerebellum.

The glutamate immunoreactivity of different cell populations was compared quantitatively in the cerebellar cortex of cat, using an antiserum raised against glutamate coupled to bovine serum albumin by glutaraldehyde. Neuronal and glial processes were identified on serial electron microscopic sections which were processed by a postembedding immunogold procedure. The surface density of colloidal gold particles was used for statistical comparison of the relative levels of glutamate in cell populations, or in different parts of the same population. The terminals of mossy and parallel fibres had significantly higher levels of glutamate immunoreactivity than Golgi cell terminals, granule cell dendritic digits, Purkinje cell dendrites or dendritic spines. Golgi cell terminals were identified by their position and GABA immunoreactivity as revealed by immunogold in serial sections. The dendritic digits of the putative glutamatergic granule cells had significantly higher glutamate immunoreactivity than did Purkinje cell dendrites and dendritic spines. Glial cell processes in the molecular layer had lower level of glutamate immunoreactivity than any of the neuronal processes. The results demonstrate that the highest levels of glutamate immunoreactivity occur in mossy and parallel fibre presynaptic terminals that are known to have an excitatory effect. This supports previous suggestions that glutamate may be a transmitter at these synapses. The measurement of the levels of putative amino acid transmitters in identified neuronal populations, or in different parts of the same population, could have wide applications in studies on the chemical neuroanatomy of the nervous system.

Animals↗
PubMed1986

Na+-dependent "binding" of D-aspartate in brain membranes is largely due to uptake into membrane-bounded saccules.

Na+-dependent "binding" of acidic amino acids in brain plasma membranes was examined by procedures similar to those employed in earlier studies, using the metabolically inert D-[3H]aspartate as a probe. The "binding" showed characteristics similar to those described before in terms of affinity (KD, 400 nM), density of sites (Bmax, 300 pmol/mg protein), sensitivity to D,L-threo-3-hydroxyaspartate, and requirement for Na+. It turned out that the "binding" represents uptake into membrane-bounded saccules (which according to the inulin and H2O spaces constituted 3.4 microliters/mg protein and comprised about 50% of the volume of the sedimented membranes), rather than binding to the transport carrier. This conclusion is based on the observations that the "binding" of D-aspartate was released by osmotic shock; was abolished by thorough washing of membranes in H2O prior to assay, which removed endogenous contents of amino acids, and could be recovered by loading the washed membranes with glutamate; was reduced by prior freezing and thawing; was low on incubation at 0 degree C; had a bell-shaped time course similar to that reported for uptake; and had a slow rate of reversal compared to the apparent KD. True binding would have considerably lower apparent Bmax than the carrier-mediated uptake. This and its likely rapid rate of dissociation would make binding to the carrier difficult to detect by the methods used up to now.

Animals↗
PubMed1986

Inhibition by K+ of Na+-dependent D-aspartate uptake into brain membrane saccules.

Na+-dependent uptake of dicarboxylic amino acids in membrane saccules, due to exchange diffusion and independent of ion gradients, was highly sensitive to inhibition by K+. The IC50 was 1-2 mM under a variety of conditions (i.e., whole tissue or synaptic membranes, frozen/thawed or fresh, D-[3H]aspartate (10-1000 nM) or L-[3H]glutamate (100 nM), phosphate or Tris buffer, NaCl or Na acetate, presence or absence of Ca2+ and Mg2+). The degree of inhibition by K+ was also not affected on removal of ion gradients by ionophores, or by extensive washing with H2O and reloading of membrane saccules with glutamate and incubation medium in the presence or absence of K+ (3 mM, i.e., IC70). Rb+, NH4+, and, to a lesser degree Cs+, but not Li+, could substitute for K+. [K+] showed a competitive relationship to [Na+]2. Incubation with K+ before or after uptake suggested that the ion acts in part by allowing net efflux, thus reducing the internal pool of amino acid against which D-[3H]aspartate exchanges, and in part by inhibiting the interaction of Na+ and D-[3H]aspartate with the transporter. The current model of the Na+-dependent high-affinity acidic amino acid transport carrier allows the observations to be explained and reconciled with previous seemingly conflicting reports on stimulation of acidic amino acid uptake by low concentrations of K+. The findings correct the interpretation of recent reports on a K+-induced inhibition of Na+-dependent "binding" of glutamate and aspartate, and partly elucidate the mechanism of action.

Animals↗
PubMed1986

Metabolism and transport of amino acids studied by immunocytochemistry.

The immunocytochemical method for demonstrating amino acids makes it possible to study accumulation and depletion of amino acids in individual tissue compartments resulting from experimental manipulations. We have incubated hippocampal slices in oxygenated Krebs solution, containing various additives, under basal conditions and during synaptic release of transmitters evoked by elevated K+ concentrations or by veratrine. Immunoreactivities for glutamate (Glu-LI), aspartate (Asp-LI), glutamine (Gln-LI), gamma-amino-butyrate (GABA-LI) and taurine (Tau-LI) have been demonstrated by specific antibodies after fixation of the slices in glutaraldehyde. Prolonged depolarisation depleted Glu-LI, Asp-LI and Gln-LI from nerve-ending-like structures. GABA-LI was less affected and Tau-LI not affected at all. The depletion of immunoreactivities could be prevented by metabolic precursors of transmitter amino acids, notably glutamine. This effect of glutamine was abolished by inhibiting glutaminase with diazooxonorleucine. Glu-LI, Asp-LI, GABA-LI and Gln-LI accumulated in astroglial cells during conditions of prolonged depolarization-induced release. The accumulation of GABA-LI in glia was strongly increased by inhibition of aminotransferases by aminooxyacetic acid. The described changes in Glu-LI were prevented by low Ca2+/high Mg2+, and promoted when the glial enzyme glutamine synthetase was inhibited by methionine sulfoximine. D-Aspartate, a metabolically inert competitive inhibitor/substrate for high affinity uptake of glutamate, inhibited the accumulation of Glu-LI in glia. The results confirm the biochemically derived theories on metabolic compartmentation in nervous tissue, and add knowledge on the dynamics of the cellular distribution of amino acids. They also indicate the possibilities offered by the present approach for studying metabolism and pharmacology at the cellular level.

Amino Acids↗
PubMed1986

Evaluation of the immunocytochemical method for amino acids.

Free amino acids can be coupled to proteins by glutaraldehyde. Rabbits immunised with a bovine serum albumin-glutaraldehyde-amino acid conjugate form antibodies that recognise similar conjugates with brain proteins in glutaraldehyde-fixed tissue. Antisera raised against conjugated GABA (gamma-aminobutyrate), glutamate, aspartate, taurine, glutamine, or glycine were tested against a variety of small molecular compounds that had been fixed by glutaraldehyde to brain protein and immobilised on cellulose ester filters for processing together with the brain sections. This system permitted closely similar conditions for testing and immunocytochemistry. After removing antibodies against the carrier used for immunisation and against cross reacting amino acid conjugates the antisera showed a high specificity. The specific nature of the antisera was corroborated by solid phase adsorption to the homologous antigens and by inhibition experiments with free amino acids and amino acid-glutaraldehyde fixation complexes. After transection of the striatonigral pathway the ipsilateral substantia nigra was almost depleted of GABA-like immunoreactivity; this observation lends additional support to the selectivity of the GABA antiserum. A semiquantitative relation was established between the concentration of amino acid before fixation in a model system and the subsequent intensity of immunostaining. Similar model experiments suggested that the conjugation of an amino acid to brain protein with glutaraldehyde, and the immunoreactivity of the conjugates, may be significantly inhibited in the presence of high concentrations of other amino compounds.

Amino Acids↗