Britta Nyström

Ischemia-Induced Shift of Inhibitory and Excitatory Amino Acids from Intra- to Extracellular Compartments

Brain ischemia was induced for 10 or 30 min by clamping the common carotid arteries in rabbits whose vertebral arteries had previously been electrocauterized. EEG and tissue content of high energy phosphates were used to verify the ischemic state and to evaluate the degree of postischemic recovery. Extracellular levels and total contents of amino acids were followed in the hippocampus during ischemia and 4 h of recirculation. At the end of a 30-min ischemic period, GABA had increased 250 times, glutamate 160 times, and aspartate and taurine 30 times in the extracellular phase. The levels returned to normal within 30 min of reflow. A delayed increase of extracellular phosphoethanolamine and ethanolamine peaked after 1–2 h of reflow. Ten minutes of ischemia elicited considerably smaller but similar effects. With respect to total amino acids in the hippocampus, glutamate and aspartate decreased to 30–50% of control while GABA appeared unaffected after 4 h of reflow. Alanine, valine, phenylalanine, leucine, and isoleucine increased severalfold. The importance of toxic extracellular levels of excitatory amino acids, as well as of high extracellular levels of inhibitory amino acids, are considered in relation to the pathophysiology of neuronal cell loss during cerebral ischemia.

Concentrations of amino acids in extracellular fluid after opening of the blood-brain barrier by intracarotid infusion of protamine sulfate

Abstract: This article evaluates the influence of an opening of the blood‐brain barrier (BBB) on compounds in brain extracellular fluid. The concentrations of amino acids and some other primary amines were determined in dialysates sampled from the right parietal cortex of rats before and after an intracarotid infusion of protamine sulfate. Extravasated plasma proteins were visualized by Evans blue/albumin and immunohistochemistry. CSF albumin— an indicator of blood‐CSF barrier opening—was quantified with immunoelectrophoresis. The brains were macroscopically edematous after 10 mg but not after 5 mg of protamine sulfate. The higher dose led to a 50% death rate. The concentrations of amino acids did not change 10 min after the BBB opening. No significant alterations in the amino acid concentrations were observed after the lower dose. The concentrations of glutamate, aspartate, GABA, glycine, taurine, and phosphoethanolamine increased significantly within 50–80 min after the infusion of 10 mg of protamine sulfate. CSF albumin levels were significantly increased 1 h after infusion. We conclude that a dysfunction of the BBB, of a degree known to induce brain edema (10 mg of protamine sulfate), significantly increases the extracellular concentration of excitatory amino acids, GABA, taurine, and phosphoethanolamine in the extracellular space.

Ammonium ion inhibition of evoked release of endogenous glutamate from hippocampal slices.

Abstract— The effect of pathophysiological levels (2‐5 mm) of ammonium chloride on the efflux of endogenous and exogenous [14C]glutamate from hippocampal slices was studied. The evoked release of glutamate which occurs dring tissue depolarization with 56 mm‐KCl was greatly reduced when the tissue had been exposed to NH4Cl for 40–80 min. This effect was seen whether or not glutamine (0.5 mm) was present in the incubation medium. The effect was completely reversible. The spontaneous efflux and the evoked release of [14C]glutamate was, on the contrary, completely unaltered after exposure of the slice to ammonium ions. Nigher (20–36 mm) amounts of NH4Cl evoked a release of [14C]glutamate from the crude mitochondrial fraction, as did high concentrations of KCl. The results are discussed in relation to the compartmentation of glutamate metabolism and the pathogenesis of hepatic coma.

Extracellular Amino Acids as Markers of Myocardial Ischemia during Cardioplegic Heart Arrest

Extracellular levels of amino acids in the myocardial interstitium are sensitive indicators of myocyte function. Lowered ATP leads to a rapid extracellular appearance of amino acids with a high intra- to extracellular concentration ratio, such as taurine and glutamate. Nitrogen fluxes are reflected by glutamine, while alanine, glycine, serine and leucine are markers of proteolysis. In addition, degradation of membrane phospholipids is reflected by other primary amines, such as phosphoethanolamine. The time course of these changes was determined before, during and after cardioplegic heart arrest. Two regions of the heart were monitored in 20 patients by means of microdialysis sampling. After only 20 min of heart arrest, extracellular taurine, glutamate and phosphoethanolamine increased transiently up to 25 times the basal level. Ten–20 min later, glutamine increased by 6 times. A doubling of alanine, glycine, serine and leucine levels took place 30 min after release of the aortic cross-clamp. After 2 h, all were at levels similar to those recorded 15–30 h later. Levels of taurine and glutamate in the anterior wall of the heart correlated significantly with those of its lateral wall. The response to surgery and heart arrest was studied in a group of patients with ischemic heart disease as well as in another group of patients, who underwent heart surgery for nonischemic reasons. The response of taurine and glutamine was significantly higher for the patients with ischemic heart disease, in spite of a shorter mean time of heart arrest. No sex differences were recorded. High levels of amino acids coincided frequently with clinical events, which were suggestive of ischemia, but were also recorded in a few patients without diagnosed events. We conclude that monitoring of extracellular amino acids is valuable for evaluation and development of cardioprotective strategies.

The neuronal environment after subarachnoid haemorrhage – correlation of amino acid and nucleoside levels with post-operative recovery

The study explores biochemically the neuronal environment adjacent to a subarachnoid haemorrhage in 11 patients after neurosurgical clipping of an arterial aneurysm. Extracellular fluid (ECF) from the rectus gyrus and subarachnoid fluid (SAF) were sampled with microdialysis probes. The concentrations of amino acids and nucleosides were monitored in 60 min samples collected over 2-4 days. The patients were 33-67 years of age. Surgery was performed 0-5 days after rupture of the aneurysm in 8 patients. One patient was operated on after 15 months. Clipping of aneurysms without prior haemorrhage was performed in two cases. Markedly elevated concentrations of the excitatory amino acid glutamate was observed in the ECF of only one patient who underwent surgery within 8 hours after the haemorrhage. Moderate glutamate elevations were seen in two patients and of aspartate in another patient. Five patients displayed periods of varying length of specifically elevated taurine concentrations in ECF or SAF. Transient periods of high concentrations of glycine and serine were seen in two patients. Even though average concentrations of all amino acids were fairly similar in the ECF and SAF, the pattern of changes vs. time differed markedly in the two compartments. Presently, we conclude that the level of consciousness in the post-operative phase was inversely related to total amino acid concentration in the ECF. Furthermore, while the ECF concentrations of taurine and glycine increased both specifically and transiently in several patients, excitatory amino acid levels were not appreciably elevated subsequent to the neurosurgical intervention.(ABSTRACT TRUNCATED AT 250 WORDS)

The contribution from the choroid plexus and the periventricular CNS to amino acids and proteins in the human CSF

During neurosurgery the freshly secreted extracellular fluid (ECF) from the choroid plexus was sampled with small pieces of application paper in three patients with intractable epilepsy. The samples were analyzed for free amino acids and for soluble proteins. The results were compared with corresponding data on extracellular fluid from the brain surface obtained with dialysis-perfusion as well as with the cerebrospinal fluid (CSF) acquired by lumbar punction. The dialysis data were calibrated against the paper results. The choroid plexus secretion had a high concentration of transthyretin as well as of an unidentified protein with an isoelectric point of 7.4. The cortical ECF exhibited high concentrations of tau-globulin and gamma-trace protein. Among the amino acids, glutamine had lower concentration in the choroid plexus secretion and higher concentrations in the ECF of the brain compared to the CSF. The amino acid derivative ethanolamine exhibited a similar pattern. This was interpreted to demonstrate that these compounds enter the CSF from the brain tissue. In contrast, alanine, serine, and taurine had a lower concentration in the CSF than in the plexus secretion which suggests that they are removed from the CSF by brain tissue.

High Levels of Glycine and Serine as a Cause of the Seizure Symptoms of Cavernous Angiomas

Abstract: Cavernous angiomas are vascular malformations that cause neurodegeneration and symptoms including epileptiform seizures, headache, and motor deficits. Following neurosurgical removal of the angiomas, patients mostly recover well and become seizure‐free. This study reports on the levels of certain amino acids in angiomas, obtained from 13 patients. Distinct zones of the angiomas were analyzed, from the thrombotic core, via gliotic, hemosiderin‐infiltrated intermediate zones, to a periphery without macroscopic abnormalities. The neurotransmitter amino acids glutamate, aspartate, and GABA as well as phosphoethanolamine displayed decreasing levels from the periphery to the core, reflecting the gradual neuronal loss. Compared with normal brain tissue, there was a marked increase in the levels of serine (fivefold), glycine (10‐fold), and ethanolamine (20‐fold) in the peripheral zone of the cavernous angiomas. The results are discussed in relation to seizures and NMDA receptor activation, neuron‐glia interactions, membrane phospholipids, and blood‐brain barrier function.

Extracellular glial fibrillary acidic protein and amino acids in brain regions of patients with subarachnoid hemorrhage-correlation with level of consciousness and site of bleeding

The extracellular fluid of two cortical regions was investigated in eight patients who underwent surgery for subarachnoid hemorrhage within 7-96 h after the rupture of an arterial aneurysm. Microdialysis samples, collected from the cortex of the ipsilateral gyrus rectus and temporal lobe for 52-127 h, were analyzed with respect to amino acids and the glial fibrillary acidic protein. In agreement with a previous study, an inverse relation was observed between total amino acid concentration and the level of consciousness. We also found that the concentration of glial fibrillary acidic protein was higher close to the bleeding site. Furthermorel we observed specific changes in the concentration of the protein and certain amino acids which coincided with clinical events such as increased intracranial pressure, vasospasm, ischemia and infarction. Finally, the concentrations of taurine may corroborate its function as an osmoequivalent. [Neural Res 1997; 19: 361-368].

Co-variation of free amino acids in human epileptogenic cortex

The concentration of free amino acids was measured in 41 surgically removed samples of human epileptogenic brain and in 7 specimens of non-epileptic brain tissue, removed during surgery for meningiomas, etc. The material was subdivided according to the neuropathological diagnosis: mild cortical dysplasia (MCD), gliosis astrocytoma infiltration and a histologically heterogeneous group. The non-tumoral epileptogenic samples had five times higher than normal concentration of ethanolamine and 50% elevated concentration of glycine. The concentration of other neurotransmitter amino acids did not differ markedly between epileptogenic and non-epileptic samples. The concentration of neurotransmitter amino acids showed a strong correlation with the enzyme neuron specific enolase (NSE) and were low in most samples with astrocytoma infiltration. On the other hand, tyrosine and leucine had higher concentrations in samples with lower NSE concentration. Factor analysis of the amino acids revealed four groups of covarying compounds in the brain samples, first, a neurotransmitter group, including aspartate, glutamate, GABA and phosphoethanolamine. Another group contained ethanolamine, glutamine, glycine and taurine. Factor analysis on corresponding extracellular amino acids showed two groups, the first being a “neurotransmitter” group, containing serine, taurine phosphoethanolamine and ethanolamine in addition to aspartate and glutamate. The other group consisted of asparagine, glycine, alanine, tyrosine, valine, phenylalanine, isoleucine and leucine.

Effect of serine and ethanolamine administration on phospholipid-related compounds and neurotransmitter amino acids in the rabbit hippocampus.

Abstract: The report concerns mechanisms for the increase of extracellular levels of ethanolamine and phosphoethanolamine in CNS regions, such as the hippocampus, in transient brain ischemia, hypoglycemia, seizures, etc. l‐Serine (2.5–10 mM), d‐serine (10 mM), or ethanolamine (10 mM) was administered for 20 min via a microdialysis tubing to the hippocampus of unanesthetized rabbits. The concentrations of primary amines were determined in the dialysates. When levels were elevated 10–100 times in the extracellular fluid, l‐serine caused a dose‐dependent increase of the concentration of extracellular ethanolamine. Ethanolamine caused a corresponding, although somewhat smaller, increase in serine levels. Furthermore, l‐serine also induced an increased concentration of phosphoethanolamine that was delayed in time relative to the peak of ethanolamine. d‐Serine was as effective as l‐serine in raising ethanolamine levels but had no effect on phosphoethanolamine. Ethanolamine, but not l‐serine, also increased extracellular glutamate/aspartate levels in an MK‐801‐dependent fashion. A similar effect, but delayed in time, was observed with d‐serine. These effects were inhibited by MK‐801. The concentrations of other amino acids were not significantly affected. The characteristics of the effects are suggestive of base exchange reactions between serine and ethanolamine and between ethanolamine and serine glycerophospholipids, respectively, in neuronal plasma membranes.