Search PubMed⌕ Search

Biomedical subjects

J Storm-Mathisen

Publications and source records attributed to J Storm-Mathisen.

At least 127 records · Page 7Linked to original sources

Taurine in the hippocampal formation of the Senegalese baboon, Papio papio: an immunocytochemical study with an antiserum against conjugated taurine.

An antiserum raised against taurine conjugated to bovine serum albumin by glutaraldehyde produced intense staining of hippocampal pyramidal neurons at the CA1/CA3 transition (including CA2) and of a small proportion of the granule cells. Strongly immunoreactive neurons were also found in a zone overlapping the second reflected blade in the hilus. Most glial cells were unlabeled.

Animals↗

Different neuronal localization of aspartate-like and glutamate-like immunoreactivities in the hippocampus of rat, guinea-pig and Senegalese baboon (Papio papio), with a note on the distribution of gamma-aminobutyrate.

Antisera were raised in rabbits against aspartate or glutamate conjugated to bovine serum albumin by glutaraldehyde. After immunosorbent purification the antisera reacted selectively with brain protein-glutaraldehyde conjugates of the respective amino acids. These results from model systems encouraged us to employ the antisera to study the distribution of free aspartate and glutamate in brain tissue. The aspartate antiserum produced intense staining of interneurons and deep hilar neurons and modest labelling of pyramidal and granular cells in the hippocampal formation of rats, guinea-pigs and baboons perfusion-fixed with glutaraldehyde. In contrast, glutamate-like immunoreactivity was generally high in pyramidal and granular cells and low in interneurons. In hippocampal slices immersion fixed in glutaraldehyde after being soaked in Krebs' solution aspartate-like and glutamate-like immunoreactivities were lost from perikarya and dendrites. The staining that remained occurred in nerve terminal-like dots and matched the distribution of the major excitatory fiber systems, except that only glutamate-like immunoreactivity, and not aspartate-like immunoreactivity, was concentrated at the site of the mossy fiber terminals, and that aspartate-like but not glutamate-like immunoreactivity occurred between the granular and pyramidal cell bodies. The present technique specifically demonstrates aspartate and glutamate in glutaraldehyde-fixed tissue. We suggest that in perfusion-fixed material the staining intensities reflect the total concentrations of the amino acids (i.e. the "metabolic pool" plus the "transmitter pool"). In immersion-fixed hippocampal slices the "transmitter pool" may be preferentially visualized.

Animals↗

Glutamate- and GABA-containing neurons in the mouse and rat brain, as demonstrated with a new immunocytochemical technique.

Antisera were raised against gamma-aminobutyric acid (GABA) or glutamate (Glu) conjugated to bovine serum albumin with glutaraldehyde. After purification, these antisera reacted strongly with fixed GABA or Glu, but not significantly with other amino acids fixed with glutaraldehyde to brain macromolecules. The antisera were used to demonstrate the distributions of Glu-like and GABA-like immunoreactivities (Glu-LI and GABA-LI) in parts of the perfusion-fixed mouse and rat brain, including the olfactory bulb, cerebral neocortex, thalamus, basal ganglia, lower brain stem, and cerebellum. The level of GABA-LI varied widely among brain regions, thus it was very high in the globus pallidus and substantia nigra and low in the bulk of the thalamus. The GABA antisera labeled nonpyramidal neurons of the neocortex, most cells of the reticular nucleus of the thalamus, medium-sized cells of the caudatoputamen, and stellate, basket, Golgi, and Purkinje cells of the cerebellum. The distribution of GABA-LI closely matched that of the GABA-synthesizing enzyme, glutamic acid decarboxylase (GAD), as revealed in immunocytochemical studies by others. However, the GABA antisera seem to be better suited than GAD antisera for demonstrating putative GABA-ergic axons. The results suggest that GABA-LI, as displayed by the present method, is a good marker of neurons thought to use GABA as a transmitter. Glutamate-like immunoreactivity was much more evenly distributed among regions than GABA-LI, but was particularly low in globus pallidus and substantia nigra and high in the cerebral cortex. Mitral cells of the olfactory bulb, pyramidal neocortical cells, and other cells assumed to use Glu or aspartate as transmitter were stained for Glu-LI, but so also were neurons that are thought to use other transmitters, such as cells in the substantia nigra pars compacta, in the dorsal raphe nucleus, and in the brain stem motor nuclei. The Glu antisera seem to reveal the "transmitter pool" as well as the "metabolic pool" of Glu in perfusion-fixed material. This report shows that it is possible by means of immunocytochemistry to display reliably the tissue contents of GABA and Glu in material that has been fixed by perfusion with glutaraldehyde.

Animals↗

GABA-containing neurons in the thalamus and pretectum of the rodent. An immunocytochemical study.

Antisera produced by immunizing rabbits with GABA conjugated to bovine serum albumin reacted, after purification, strongly with GABA fixed with glutaraldehyde to rat brain macromolecules, but insignificantly with other fixed amino acids (Storm-Mathisen et al. 1983). Sections through the diencephalon of perfusion-fixed mouse and rat brains showed a highly selective labeling pattern after incubation with these antisera. All cells of the reticular nucleus appeared to be stained. Smaller proportions of stained perikarya occurred in the dorsal and ventral subdivisions of the lateral geniculate body, in the medial geniculate body, in the lateroposterior nucleus, and in all nuclei of the pretectum. Labeled cell bodies were only rarely encountered in the ventrobasal complex, and were not found in the anterior and medial groups of thalamic nuclei. Stained axons were particularly concentrated in the ventrobasal complex, and in the stria medullaris, stria terminalis and inferior thalamic peduncle. The arrangement and density of labeled bouton-like dots varied markedly among nuclei, the highest densities occurring in the paraventricular and parataenial nuclei, and in the ventral subdivision of the lateral geniculate body. The mean staining intensity of the thalamic neuropil was lower than that of nearby structures, such as the hypothalamus and zona incerta. The present results on direct immunocytochemical detection of GABA are consistent with, and extend, data from immunocytochemical studies of the GABA-synthetizing enzyme, glutamic acid decarboxylase.

Animals↗

Uptake of D-aspartate and L-glutamate in excitatory axon terminals in hippocampus: autoradiographic and biochemical comparison with gamma-aminobutyrate and other amino acids in normal rats and in rats with lesions.

High affinity uptake sites for 3H-labelled amino acids were studied in synaptosome-containing homogenates processed biochemically or in surface autoradiograms of incubated slices of hippocampus. D-aspartate and L-glutamate had apparently identical distributions. In normal rat hippocampus the highest uptake was in the terminal fields of axons from the pyramidal cells of regio inferior and hilus fasciae dentatae, while there was a moderate uptake in the terminal fields of the medial and lateral perforant paths, slight uptake in the mossy fibre layer and no uptake in the terminal fields of the basket cells. Uptake sites for gamma-aminobutyrate were concentrated in the latter fields, and in the most superficial cortical layers. The present method shows no uptake in cell bodies. The uptake activities were strongly inhibited by recognized blockers of (neuronal) high affinity uptake of glutamate or gamma-aminobutyrate. Autoradiographically, several other amino acids showed negligible uptake. The uptake of D-aspartate was reduced by 80% in stratum oriens and stratum radiatum of regio superior 4-14 days (70% at 3 days) after transection of the afferent pyramidal cell axons from the ipsi-and contralateral regio inferior. The reduction was in the number of uptake sites, not in their affinity. Uptake of gamma-aminobutyrate was not reduced. Lesions affecting regio superior caused a loss of D-aspartate uptake in subiculum at a site known to receive hippocampal afferents. Autoradiographically, the uptake of D-aspartate was strongly reduced in the inner zone (i.e. the target zone), but increased in the middle zone of the dentate molecular layer after lesions of the hilus fasciae dentatae. At 4 days and longer after transection of the entorhinal afferents, there was a conspicuous reduction of D-aspartate and L-glutamate uptake in the target zones of both the medial and lateral contingent of these fibres. In the same animals, the terminal zone of afferents from hilus fasciae dentatae had an increased radioactivity and was slightly wider than normally. Concomitantly, the gamma-aminobutyrate uptake was increased in the target zones of the degenerating perforant path fibres. The results demonstrate that uptake sites for D-aspartate and L-glutamate are highly localized in axon terminals of regio inferior pyramidal cells and in perforant path afferents. The latter category of terminals has a lower density of acidic amino acid uptake sites than the former. Uptake sites for gamma-aminobutyrate are localized in terminals of intrinsic neurones, including the axosomatic terminals of basket cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Absorption↗

Lesions of Schaffer's collaterals in the rat hippocampus affecting glutamate dehydrogenase and succinate dehydrogenase activity in the stratum radiatum of CA 1. A study with special reference to the glutamate transmitter metabolism.

Lesions of CA 3 derived axons, comprising Schaffer's collaterals, were carried out in order to destroy their presumably glutamatergic nerve endings within CA 1. After a survival time of 20 days part of the stratum radiatum of CA 1 displayed statistically significant reduction of histochemically demonstrable glutamate dehydrogenase and succinate dehydrogenase activity by about 19 and 25 per cent, respectively, whereas alpha-glycerophosphate dehydrogenase was not affected. These findings are consistent with current biochemical and histochemical results on the relation between several glutamate producing enzymes and glutamatergic structures suggesting that glutamate dehydrogenase plays a major role in glutamate transmitter metabolism.

Animals↗

Retrograde transport of D-[3H]aspartate in thalamocortical neurones.

D-[3H]aspartate (D-Asp) injected into the sensorimotor cortex of the rat resulted in retrograde labelling of thalamic neurones, mainly in the intralaminar and ventromedial nuclei, and the posterior complex. Few cells in the ventrolateral nucleus and ventrobasal complex were labelled. The results show that certain thalamocortical neurones can take up and axonally transport the false transmitter D-Asp, indicating that they might use glutamate or aspartate as transmitter(s).

Animals↗

First visualization of glutamate and GABA in neurones by immunocytochemistry.

Immunocytochemical methods for peptides and serotonin have greatly advanced the study of neurones in which these substances are likely to be transmitters. Such direct techniques have not so far been available for the amino acid transmitter candidates. We report here the selective immunocytochemical visualization of the putative transmitters glutamate (Glu) and gamma-aminobutyrate (GABA) by the use of antibodies raised against the amino acids coupled to bovine serum albumin (BSA) with glutaraldehyde (GA). The tissue localizations of Glu-like and GABA-like immunoreactivities (Glu-LI and GABA-LI) matched those of specific uptake sites for Glu and GABA, and, in the case of GABA-LI, also that of the specific marker enzyme glutamic acid decarboxylase (GAD). Thus, GABA-LI was located in what are believed to be GABAergic inhibitory neurones, whereas Glu-LI was concentrated in excitatory, possibly glutamatergic neurones. Preliminary electron microscopic observations suggest that the transmitter amino acids are significantly concentrated in synaptic vesicles.

Animals↗

The corticopontine projection: axotomy-induced loss of high affinity L-glutamate and D-aspartate uptake, but not of gamma-aminobutyrate uptake, glutamate decarboxylase or choline acetyltransferase, in the pontine nuclei.

The corticopontine fibres were severed in the crus cerebri in rats and mice by a stereotaxically operated retractable wire-knife. The pontine nuclei were microscopically dissected from fresh slices of rats and synaptosome-containing homogenates were prepared. The high affinity uptake of radiolabelled L-glutamate (L-Glu) and D-aspartate (D-Asp) was heavily reduced five days after the lesions. The uptake was further reduced after bilateral (-75% for D-Asp and -65% for L-Glu) than after unilateral lesions (-55% for D-Asp and -45 to 50% for L-Glu on the lesioned side.) The molar ratio of the uptakes of D-Asp and L-Glu was consistently lower in pons after transection of the cortical afferents than normally (-28% after bilateral lesions). gamma-Aminobutyrate uptake and glutamic acid decarboxylase were not changed. Choline acetyltransferase was increased (+53%) after unilateral lesions, but not altered after bilateral lesions. Autoradiograms of slices from mice, incubated with tritium-labelled amino acids and fixed in glutaraldehyde, showed high affinity uptake sites for D-Asp to be enriched in the pontine nuclei, compared to neighbouring structures. After partial lesion of the crus cerebri the uptake was reduced in the area with degenerated corticopontine afferents. gamma-Aminobutyrate uptake sites were relatively less concentrated in the pontine nuclei than D-Asp uptake sites. The results indicate, along with the previous demonstration of Ca-dependent K-induced release of D-[3H]aspartate from the corticopontine terminals, that glutamate and/or aspartate may be transmitters in this pathway. The results also suggest that acidic amino acid uptake sites may differ in their relative transport rates for aspartate and glutamate.

Animals↗

In vivo high-affinity uptake and axonal transport of D-[2,3-3H]aspartate in excitatory neurons.

D-[2,3-3H]aspartate ([3H]D-Asp) at microM concentrations in Krebs' solution was infused intracerebrally in rats, mice and hamsters. Neuropil sites in the hippocampal formation, septum and neostriatum, known to receive excitatory nerve inputs with glutamate and aspartate as putative transmitters, showed strong autoradiographic labeling after intraventricular infusions. There was evidence for retrograde axonal transport to pyramidal cell bodies in hippocampus CA3 and neocortex. Infusions into the hilus fasciae dentatae led to anterograde axonal transport of [3H]D-Asp in the mossy fibers.

Animals↗

K+-evoked Caa+-dependent release of D-[3H]aspartate from terminals of the cortico-pontine pathway.

Pontine nuclei dissected from rat brain slices released previously accumulated D-[3H]aspartate (D-Asp) and [14C]gamma-aminobutyrate (GABA) Ca-dependently when exposed to 50 mM K. These efflux rates were substantially increased by including 0.5 mg/ml bovine serum albumin in the superfusion fluid. Degeneration of the cortico-pontine fibres 5 days after cutting the crus cerebri caused an 80% reduction in the fractional rate of Ca-dependent D-Asp release and a 60% reduction in uptake. The fractional rate of GABA efflux was significantly less reduced than that of D-Asp efflux, and GABA uptake was nearly unchanged.

Animals↗

Glutamate in hippocampal pathways.

The three consecutive main excitatory neurone projections in the hippocampal formation (the perforant path, the mossy fibres and the pyramidal cell axons) all show acidic amino acid uptake in their terminals. This is in accordance with other data pointing to the transmitter function of glutamate and/or aspartate in the perforant path and pyramidal cell axons. In the mossy fibres the role of these amino acids has so far not been determined. The localization of glutamate and aspartate uptake in the excitatory axon systems contrasts with the localization of GABA uptake in the axons of the inhibitory basket cells and other short axon neurones.

Animals↗

Tentative localization of glutamergic and aspartergic nerve endings in brain.

1. The locations of the high affinity uptakes of glutamate, aspartate and GABA were studied autoradiographically and microchemically in slices of hippocampus and septum in vitro. 2. In hippocampus the distributions of the uptake sites for glutamate and aspartate were very similar, with much higher uptake in zones containing pyramidal cell terminals than in other zones. A reciprocal distribution was found for GABA uptake, which was in agreement with that of GAD. 3. Cutting pyramidal cell axons to CAl reduced the uptake of aspartate and glutamate in the target area in CAl by 80%. 4. Autoradiographically the uptake of aspartate was very high in the dorsal part of the lateral septum, moderately high in nucleus accumbens septi and neostriatum, and very low in the medial septum. GABA uptake was lower in the medial than in the lateral septum, but very high in a narrow transitional zone and in the insula Cajella magna. 5. Transecting the axons from hippocampus and subiculum to septum, gave a 70% reduction in the uptakes of aspartate and glutamate in the lateral septum, but no reduction in the medial septum. 6. Literature data on uptake, content and release of glutamate and aspartate in nerve endings in brain are briefly reviewed.

Animals↗

Localization of tissue thromboplastin in the human brain.

Monospecific antisera against the purified protein component of tissue thromboplastin (apoprotein-III) from human brain have been raised in goats and rabbits. The antisera neutralized tissue thromboplastin prepared from brain, thyroid gland and pulmonary tissue, indicating that apoproteins in the various preparations cross-reacted immunologically and therefore were similar or identical. Comparison of the activities of tissue thromboplastin preparations from 34 different areas of the brain demonstrated a characteristic distribution pattern and a wide range of activities. White and grey matter from the same areas had similar activities. Bulbus and tractus olfactorius, medulla oblongata, corpus pineale, hippocampus and hypothalamus contained 160-270% of the average activity, whereas cerebellum globus pallidus, nucleus ruber and substantia nigra contained 30-60%. The distinct distribution pattern was unrelated to tissue vascularization, and may suggest that apoprotein-III could serve other functions, apart from the coagulation of blood. The predominance in phylogenetically older brain regions would suggest that it represents a primitive or fundamental feature.

Animals↗