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Biomedical subjects

A Hamberger

Publications and source records attributed to A Hamberger.

At least 109 records · Page 6Linked to original sources

S-100 alpha-like immunoreactivity in tubules of rat kidney. A clue to the function of a "brain-specific" protein.

The localization of the alpha and beta subunits of S-100 protein was studied in normal tissue where the identification of three subclasses of S-100 containing cells was derived: i) cells that contain both alpha and beta subunits; ii) cells that contain only the alpha subunit; and iii) cells that contain only the beta subunit. In this study monospecific antibodies against the S-100 alpha and beta subunits were used to characterize the S-100-like immunoreactivity in the rat kidney: Certain cells in the distal nephron, i.e., the connecting piece, collecting ducts, and the thin limb of Henle's loop, contained S-100 alpha immunoreactivity. Proximal tubules, the thick ascending limb of Henle's loop, the distal tubules, and the juxtaglomerular apparatus were negative. No S-100 beta immunoreactivity was found in kidney tubules. However, Schwann cells of renal pelvic nerves contained S-100 beta immunoreactivity. The presence of S-100 alpha antigen in certain cells of the kidney gives further support to the assumption that the alpha subunit of S-100a is related to cells that are highly involved in pH, electrolyte, and water regulation.

Animals↗

A role for taurine in the maintenance of homeostasis in the central nervous system during hyperexcitation?

Employing the brain dialysis technique, we demonstrate that the aspartate congener N-methyl-D,L-aspartic acid (NMA) stimulates the release of endogenous taurine in vivo in a Ca2+-dependent manner. Furthermore, exogenous taurine (1-10 mM) inhibits the NMA-induced Ca2+ influx into intracellular compartments. This suggests that the extracellular taurine concentration may control Ca2+ movement and thereby provide a homeostatic mechanism in situations of excessive excitation.

Animals↗

Veratridine-induced release in vivo and in vitro of amino acids in the rabbit olfactory bulb.

Free amino acids were studied in the olfactory bulb of the rabbit during basal conditions and veratridine-induced depolarization, in vitro with a tissue slice preparation and in vivo with a perfusion-dialysis technique. In vivo, basal extracellular concentrations of GABA, beta-alanine and aspartate were low, while glutamine showed the highest level. The basal steady-state concentration ratio between the total tissue pool of free amino acids and amino acids in the extracellular fluid was high for GABA, aspartate and glutamate, while low for glutamine and other 'non-transmitter' amino acids. Veratridine induced a marked TTX-sensitive release of GABA (40-50 times the control) both in vivo and in vitro. In vivo, the GABA release showed a peak during the first minutes of veratridine perfusion. The TTX-sensitive release of aspartate and glutamate, on the other hand, was approximately 5 times higher in vitro than in vivo. Furthermore, a prolonged response to veratridine was seen for glutamate and aspartate in vivo consisting of an early peak, followed by a sustained release. Taurine showed a time-delayed veratridine response, both in vivo and in vitro, whereas glutamine displayed a slow, TTX-sensitive decrease. No effect of veratridine was seen on beta-alanine or carnosine-threonine levels.

Amino Acids↗

Glial contribution to amino acid content and metabolism of the deafferented dentate gyrus.

The time course of tissue content and evoked release of endogenous amino acids was analyzed in the partially deafferented dentate gyrus of the rat hippocampus 2-24 days following unilateral lesion of the perforant path. Amino acids in tissue extracts and perfusates were determined after precolumn derivatization and hplc separation. The astrocytic glial cell reaction was monitored with immunohistochemistry of S-100. The tissue content of glutamate decreased significantly on the lesioned side, whereas only a moderate reduction in taurine, aspartate, and alanine occurred. Glutamine was significantly elevated at 7 days. The evoked efflux of glutamate was reduced at 2 and 7 days, whereas no change was seen at longer survival periods. The evoked release of GABA and aspartate increased on the denervated side after 12 and 24 days. The rate of carbon utilization into amino acid pools was followed with 14C-glucose and 14C-acetate. The incorporation of acetate showed a peak 2-9 days following lesion, which paralleled in time the hypertrophic glial cells. The incorporation of glucose decreased during this period. The metabolic events are discussed in relation to the morphological changes in synapses and glial cells.

Acetates↗

Extra- and intracellular amino acids in the hippocampus during development of hepatic encephalopathy.

Fulminant hepatic failure was induced in rabbits by intravenous administration of galactosamine hydrochloride. The animals were sacrificed after 45 h and the hippocampus analyzed for free amino acids. In addition, free amino acids were measured in plasma and in the extracellular fluid of the hippocampus 20, 30 and 45 h after galactosamine injection. The extracellular fluid compartment was analyzed by slow perfusion of a thin dialysis tube which was implanted in the hippocampus one day prior to galactosamine administration. The amino acid concentration in the extracellular fluid agreed fairly well with that of the cerebrospinal fluid in the control situation. During development of hepatic failure, the plasma concentration of all amino acids increased. The changes in extracellular amino acids were smaller, except for phosphoethanolamine and glutamate. The concentration ratio intra/extracellular amino acids decreased in the hippocampus for amino acids with a normally high concentration gradient.

Amino Acids↗

Inhibition by verapamil of ischemic Ca2+ uptake in the rabbit hippocampus.

In the study described here a perfusion-dialysis system was used for recording ionic changes in the hippocampus during ischemia. Local perfusion with a Ca2+ entry blocker (verapamil) inhibited the fall in extracellular Ca2+ concentration which occurs in ischemia. The data consequently suggest that intracellular Ca2+ accumulation during ischemia can be blocked. A Ca2+ antagonist such as verapamil may thus improve post-ischemic neuronal recovery via a direct effect on parenchymal cells in addition to the previously suggested vascular effects.

Animals↗

A possible interrelationship between extracellular taurine and phosphoethanolamine in the hippocampus.

The effect of guanidinoethane sulfonic acid (GES), an inhibitor of taurine uptake, was examined with respect to endogenous amino acids in the hippocampus of the freely moving rabbit. GES increased the extracellular levels of both taurine and phosphoethanolamine (PEA), other amino acids being unaffected. However, long-term oral administration of GES selectively reduced endogenous taurine levels. The effect of GES on PEA appeared to be a consequence of the elevated extracellular taurine as exogenously administered taurine per se increased PEA levels in the extracellular space. The findings are discussed in conjunction with the proposed membrane-stabilizing effects of taurine.

Animals↗

Effect of ambient temperature on rectal temperature in normal and malnourished rats during early postnatal development.

Information on body temperature is frequently lacking in metabolic studies on normal and malnourished suckling rats although differences in their thermoregulatory efficiency may be expected. The aim of the present study was to evaluate the effect of different ambient temperatures on the rectal temperature of litters of normal and protein deprived rats to establish the ambient temperatures at which a normal rectal temperature can be expected at various postnatal ages. The offspring of normal rats and rats subjected to a dietary reduction of 50% of the protein intake were examined 10 days to 30 days postnatally. The pups were transferred from the nest cage to an incubator: the rectal temperature was measured immediately after transfer to the incubator and 30 and 60 min later at ambient temperatures of 35, 30 and 25 degrees C. Up to 15 days of age control pups showed a fall in rectal temperature after the short exposure to room temperature (22 degrees C) needed for transfer. A similar fall of rectal temperature was recorded in the malnourished pups up to 25 days age. The effects of the ambient temperatures on the malnourished pups indicated a delay in the control of body temperature of some 10 days. This is in agreement with a previous report on semi-starved rats (Heim & Szelenyi 1965). The conditions for the maintenance of a normal rectal temperature in litters of normal and malnourished suckling rats separated from their mother are out-lined.

Age Factors↗

Inhibition of cholinergic response by taurine in frog isolated skeletal muscle.

Carbamylcholine-stimulated contractions of isolated frog gastrocnemius muscle were inhibited in a non-competitive fashion by 5-25 mM taurine. Taurine had no effect on the resting length of the muscle. Caffeine-induced contractures were unaffected by taurine which indicates that the sarcoplasmic reticulum is not an important site of action for taurine. A possible functional role for taurine in skeletal muscle is discussed.

Animals↗

Differential localization of "brain-specific" S-100 and its subunits in rat salivary glands.

In the rat, the S-100 antigens in the submandibular gland were found to be immunochemically identical with those in the brain (glial cells) when compared using crossed immunoelectrophoresis. Specific antibodies against the S-100a non-beta and against the S-100 beta subunit were prepared from antibodies against crude S-100 protein and from S-100 components (S-100a and b) by affinity chromatography. In the rat salivary glands a differential distribution of subunit immunoreactivity was clearly evidenced using indirect immunofluorescence. Certain intercalated duct cells of the submandibular gland as well as Schwann cells contained the S-100 beta subunit immunoreactivity exclusively, while other duct cells in parotid, submandibular, and sublingual glands contained S-100a non-beta subunit immunoreactivity. Both subunits were present in astrocytes and ependymal cells. The immunocytochemical localization of alpha and beta subunits is a promising technique for the classification of various types of S-100-containing cells.

Animals↗

Dihydrokainic acid affects extracellular taurine and phosphoethanolamine levels in the hippocampus.

The specificity of the inhibitory effect of dihydrokainic acid on the high-affinity glutamate uptake was tested in an in vivo system. The rabbit hippocampus was perfused with Krebs--Ringer bicarbonate medium to which dihydrokainic acid was added (final concentrations 1 and 5 mM) with or without depolarizing potassium concentrations. Dihydrokainate elevated the levels of glutamate, taurine and phosphoethanolamine. This response was partly similar to that observed after high potassium administration. The selectivity of dihydrokainic acid is thus not restricted to glutamate, and its value as a tool in the study of glutamatergic transmission is discussed.

Animals↗

Effects of in vivo administration of kainic acid on the extracellular amino acid pool in the rabbit hippocampus.

The effect of local administration of kainic acid in the rabbit hippocampus was studied; the hippocampus was perfused continuously in the freely moving animal with an implanted 0.3-mm dialysis fiber. The pattern of endogenous amino acids in the perfusate, reflecting extracellular amino acids, was monitored with liquid chromatography separation and fluorimetric detection of amino acid derivatives. Kainic acid was included in the perfusion medium for up to 70 min at 0.1-1.0 mM and, with time, induced epileptiform activity. Endogenous glutamic acid, taurine, and phosphoethanolamine levels were increased selectively at the lower perfusion concentrations of kainic acid. Long perfusion periods with higher concentrations increased the levels of virtually all amino acids. Perfusion of the hippocampus with depolarizing concentrations of potassium gave an amino acid response partly similar to that seen with kainic acid treatment. However, one notable difference between the two responses was that the extracellular concentration of glutamine, although not influenced by kainic acid, was significantly decreased after high potassium concentrations. These results confirm previous notions that kainic acid has a primarily excitatory effect, one manifestation of this effect being the release of glutamic acid.

Amino Acids↗

Regulation of glutamate biosynthesis and release in vitro by low levels of ammonium ions.

The content and release of endogenous amino acids from isolated rat hippocampal slices were measured. The tissue was perfused with control media and pulsed with high potassium media in order to induce synaptic release. Pathophysiological concentrations of ammonium ions (3--5 mM) were added to the control medium for 60 min prior to the induced release. Amino acids belonging to the putative transmitter group were released extensively during potassium perfusion and, except for glutamate, even after ammonium ion perfusion. The spontaneous secretion of glutamate increased, however, slowly after the addition of ammonia. The incorporation of 14C from radiolabelled glucose and acetate into the amino acid fraction was studied in the presence of ammonia-containing media. Glucose was utilized to a moderately increasing extent, but acetate-derived radioactivity was strikingly decreased in the amino acid fraction during ammonia perfusion. The decreased acetate incorporation into amino acids was mainly due to an inhibition by ammonia of the accumulation of acetate by the CNS tissue.

Acetates↗

Effect of undernutrition on DNA and RNA synthesis in subcellular fractions from different regions of the developing rat brain.

The effect of undernutrition on the incorporation of [methyl-3H] thymidine into DNA and of 5-[3H] uridine into RNA of cerebral hemispheres, cerebellum, and brain stem was studied in vivo and in vitro in rats. The labeling of DNA from nuclei and mitochondria and of RNA from nuclei, mitochondria, microsomes, and soluble fractions, was also measured in vitro. The results demonstrate that nucleic acid synthesis is impaired and delayed during undernutrition. Specific effects were observed for the different brain regions and subcellular fractions: at 10 days nuclear and mitochondrial DNA and RNA synthesis was impaired, whereas at 30 days only the mitochondrial nucleic acid synthesis was affected. The delay of DNA and RNA labeling, caused by undernutrition, was most evident in the cerebellum, probably due to its intense cell proliferation during postnatal development. The specific sensitivity of mitochondria as compared to other subcellular fractions, may by due to the intense biogenesis and/or turnover of nucleic acids in brain mitochondria not only during postnatal development, but also in the adult animal.

Animals↗

The influence of protein deprivation during development on amino acid incorporation into cerebral proteins.

The incorporation of 3H-valine into protein of cerebrum, liver, and muscle was studied in rats fed a low protein diet. Animals, 15-50 days old, were used to cover the period during and after the most rapid phase of cerebral protein synthesis. The precursor was administered in doses resulting in brain concentrations of free valine in the 0.1-0.3 mM range. Two series of experiments were run and the injected dose varied for comparison of protein synthesis at different concentrations of precursor valine. By concomitant amino-acid analysis it was shown that the concentration of other amino-acids in the brain was not changed by the injected valine. The changes in the amino-acid pool with age in control rats were in agreement with previous findings. Only small alterations were seen in the amino acid pool as an effect of protein deprivation, the results being consistent with those of some previous reports. Uptake of valine into the brain appeared to be decreased only in 15-17-day-old protein-deprived rats. Valine incorporation into cerebral protein was decreased in the youngest age group but no effect was seen in older animals. Liver protein incorporation was slightly decreased by low protein diet but a marked reduction was observed for muscle protein. The changes in brain protein synthesis during normal development were in agreement with previous studies and differed qualitatively from those in liver and muscle.

Aging↗

Effects of low ammonia levels on NAD(P)H levels and glutamate secretion during calcium-dependent depolarization of CNS slices.

Simultaneously with an evoked release of endogenous glutamate, redox changes in NAD(P)H levels occur as a response to electrical or chemical stimulation of the isolated CNS tissue. While electrical-field stimulation induces a transient increase in tissue NAD(P)H, KCl stimulation produces a decrease in NAD(P)H. One possible interpretation for this difference is that elevated KCl induced glutamate release from larger cell populations, including glia, while electrical stimulation might have a more neuro-specific action. In the present report, it is shown that the electrically or biochemically evoked release of endogenous glutamate is strongly inhibited by ammonium ions at 3--5 mM in the hippocampus and frontal cortex. At the same time, ammonium ions inverted the NAD(P)H response of the tissue during electrical stimulation, making both electrical and KCl depolarization induce a decrease in NAD(P)H. There was no effect of ammonium ions per se on the NAD(P)H levels, and to obtain the effects on both NAD(P)H and glutamate release, the tissue had to be exposed for 40--60 minutes to ammonium ions. The results are interpreted to indicate that ammonium ions influence regulatory control mechanisms in cell populations in the tissue slice that secretes glutamate in response to depolarization.

Ammonia↗