Anke M. Mans

Regional Blood‐Brain Barrier Permeability to Amino Acids After Portacaval Anastomosis

Abstract: The influx of phenylalanine, tryptophan, leucine, and lysine across the blood‐brain barrier of individual brain structures was studied in rats 7–8 weeks after a portacaval shunt or sham operation. The method involved a brief infusion of labeled amino acid in tracer quantity and quantitative autoradiography. The clearance rates of phenylalanine, tryptophan, and leucine were increased in proportion to each other in every region examined, but not by the same factor. Tryptophan clearance increased the most (about 200%) and leucine the least (about 30%), compared with phenylalanine (about 80%). This was unexpected, as all three amino acids are believed to be transported by the same mechanism. The changes were most marked in several limbic structures and the reticular formation, whereas the hypothalamus was least affected. Plasma clearance of lysine was decreased in all areas by about 70%. Since the circulating lysine concentration was decreased by 13%, the actual rate of lysine influx was even more reduced. The results demonstrate specific alterations in two different amino acid transport systems. The resulting excess brain neutral amino acids, some of which are neurotransmitter precursors, as well as reduced basic amino acid availability, may be of etiological significance in hepatic encephalopathy.

Glucose Availability to Individual Cerebral Structures Is Correlated to Glucose Metabolism

Regional cerebral glucose influx was measured using quantitative autoradiography after the intravenous infusion of [2‐14C]glucose for a period of 10 or 20 s. Glucose influx varied considerably among structures over an almost threefold range. When compared with rates of regional glucose utilization, a significant correlation by region was found between glucose influx and utilization, demonstrating that the glucose supply to individual cere bral structures is closely matched to their metabolic needs.

TRYPTOPHAN TRANSPORT ACROSS THE BLOOD-BRAIN BARRIER DURING ACUTE HEPATIC FAILURE

Abstract— Tryptophan transport across the blood‐brain barrier was studied using a single injection dual isotope label technique, in the following three conditions: normal rats, rats with portacaval shunts, and rats with portacaval shunts followed 65 h later by hepatic artery ligation. In both normal rats and those with acute hepatic failure the tryptophan transport system was found to be comprised of two kinetically distinct components. One component was saturable and obeyed Michaelis‐Menten kinetics (normal: Vmax= 19.5 nmol.min−1.g−1. Km= 113 μM; hepatic failure: Vmax, = 33.8 nmol.min−1.g−1, Km= 108 μM), and the second was a high capacity system which transported tryptophan in direct proportion to concentration over the range tested (normal: K= 0.026 ml.min−1.g−1; hepatic failure: K= 0.067 ml.min−1.g−1). Since the saturable low capacity component transports several neutral amino acids, and their collective plasma concentration is high in relation to the individual Kms, tryptophan transport by this component is reduced by competitive inhibition under physiological conditions. Thus it was calculated that in normal rats approx 40% of tryptophan influx occurs via the high capacity system. During acute hepatic failure transport via both components was increased substantially, approximately doubling the rate of tryptophan penetration of the blood‐brain barrier at all concentrations tested. The contribution by the high capacity component became even more significant than in normal rats, accounting for about 75% of all tryptophan passage from plasma to brain. Brain tryptophan content was 29.9 nmol/g in normal rats and rose to 45.2 nmol/g in rats with portacaval shunts and 50.5 nmol/g in those with acute hepatic failure, correlating with the increased rate of tryptophan transport. In a previous study we found that plasma competing amino acids were greatly increased during acute hepatic failure. Calculations predict that these increased concentrations would cause a reduction in tryptophan transport by the low capacity system. However, because of the increase in the rate of transport by the high capacity component, net tryptophan entry across the blood‐brain barrier was actually increased. This increased rate of transport clearly contributes to the increased content of brain tryptophan found during hepatic failure.

Portacaval Anastomosis: Brain and Plasma Metabolite Abnormalities and the Effect of Nutritional Therapy

Abstract: Rats with portacaval shunts were used as a model of hepatic encephalopathy and compared to sham‐operated controls. First, the changes in intermediary metabolites and amino acids in blood and whole brain were characterized and found to be similar at 4 and 7 weeks after shunting. Second, the effects of nutritional therapy on selected metabolites and tryptophan transport into brain were assessed in rats 5 weeks after surgery. Ordinary food was removed and the rats were treated with glucose given either by mouth or intravenously, or intravenous glucose plus branched chain amino acids. Several abnormalities in plasma amino acid concentrations were reversed by treatment. The abnormally high brain uptake index of tryptophan, a consequence of portacaval shunting, was not lowered by any of the treatment regimens; it was even higher in the groups given glucose by mouth and glucose plus amino acids. Calculated competition for entry of tryptophan, phenylalanine, and tyrosine into brain was unchanged (glucose plus amino aicds), or reduced (glucose alone). Brain glutamine content was brought to near normal by all treatments. Infusion of glucose plus branched chain amino acids normalized brain content of tryptophan, phenylalanine, and tyrosine, even though the brain uptake index of tryptophan was higher in this group. Thus, partial or complete reversal of several abnormalities found after portacaval shunting was achieved by removal of oral food and administration of glucose. The addition of branched chain amino acids to the glucose infusion restored brain content of three aromatic amino acids to near normal, by a mechanism which appeared to be unrelated to transport across the blood‐brain barrier.

Intermediary Metabolism of Carbohydrates and Other Fuels

The central nervous system (CNS) is composed of many cell types, each with different functions and metabolic requirements, These are combined into a variety of structures which may be just as different metabolically as they are anatomically and functionally. Although considerable progress has been made in the separation and study of these different cell types (see Volume 1 of this Handbook), results from such studies may not necessarily reflect cerebral metabolism in vivo. It is, after all, the interaction of the various cell types that determines the unique character of cerebral function and metabolism. When j the cells are separated, this aspect is lost to a greater degree in brain than in other organs which do not rely as heavily on intercellular communication. In view of these considerations, the emphasis of this chapter is on studies conducted in vivo.

The influence of ketamine on regional brain glucose use

The purpose of this study was to determine the effect of different doses of ketamine on cerebral function at the level of individual brain structures as reflected by glucose use. Rats received either 5 or 30 mg/kg ketamine intravenously as a loading dose, followed by an infusion to maintain a steady-state level of the drug. An additional group received 30 mg/kg as a single injection only, and was studied 20 min later, by which time they were recovering consciousness (withdrawal group). Regional brain energy metabolism was evaluated with (6-14C) glucose and quantitative autoradiography during a 5-min experimental period. A subhypnotic, steady-state dose (5 mg/kg) of ketamine caused a stimulation of glucose use in most brain areas, with an average increase of 20%. At the larger steady-state dose (30 mg/kg, which is sufficient to cause anesthesia), there was no significant effect on most brain regions; some sensory nuclei were depressed (inferior colliculus, –29%; cerebellar dentate nucleus, –18%; vestibular nucleus, –16%), but glucose use in the ventral posterior hippocampus was increased by 33%. In contrast, during withdrawal from a 30-mg/kg bolus, there was a stimulation of glucose use throughout the brain (21–78%), at a time when plasma ketamine levels were similar to the levels in the 5 mg/kg group. At each steady-state dose, as well as during withdrawal, ketamine caused a notable stimulation of glucose use by the hippocampus.

Regional Cerebral Glucose Utilization in Rats with Portacaval Anastomosis

Regional cerebral glucose utilization was measured using [2‐14C]glucose in rats with an end‐to‐side portacaval anastomosis. The experiments were conducted in two groups of rats 4 to 8 weeks after portacaval shunting was established. One group was paralyzed and given N2O:O2 (70:30), whereas the other was conscious, unstressed, and unaware of the experiment. In both groups the rate of glucose utilization was decreased in almost all brain structures by an average of 20% after portacaval shunting. The results showed definitively that cerebral energy metabolism was reduced at a time when there were no obvious neurological abnormalities.

Brain monoamines after portacaval anastomosis

Norepinephrine, dopamine, and serotonin, as well as the serotonin metabolite, 5-hydroxy-indoleacetic acid, were measured in whole-brain extracts from rats with a portacaval shunt or sham operation. Norepinephrine, serotonin, and 5-hydroxyindoleacetic acid were significantly higher after shunting. There was no difference in dopamine. The results support the idea that brain indole metabolism is increased during chronic hepatic encephalopathy. However, they provide evidence against suggestions that hepatic encephalopathy in general is accompanied by a shortage in the whole-brain content of the catecholamines norepinephrine and dopamine.

Regional brain glucose utilization in rats during etomidate anesthesia.

The influence of etomidate on regional cerebral function as reflected by regional cerebral glucose utilization (rCMRGlc) was studied. Three experiments were performed. In the first, rats had both left femoral vessels cannulated and were placed in restraining cages. Etomidate was infused intravenously (12 mg/kg) at a rate of 6 mg · kg−1 · min−1. This large dose had a modest effect on blood pressure and heart rate, which could be explained by the elimination of stress in restrained rats, and no effect on body temperature, PaO2, PaCO2, or pH. A second group of rats were used to determine the effect of etomidate on the ratio of brain glucose to plasma glucose, which is necessary for calculating rCMRGlc. In the third experiment rCMRGlc was measured in unstressed rats. The rats were anesthetized with an intravenous dose of 1, 2, 6, or 12 mg/kg etomidate infused at a rate of 6 mg · kg−1 · min−1. Etomidate had a marked effect on glucose consumption in many, but not all, cerebral structures. The forebrain (telencephalon and diencephalon) was most affected (–25% to –35%) while the hindbrain was minimally affected. There was no demonstrable dose dependency; 1 mg/kg depressed rCMRGlc as much as 12 mg/kg. The pattern of rCMRGlc depression is in accord with the minimal effects observed on physiologic variables and similar to that caused by the steroid anesthetic Althesin,® although the depression seen was not as severe. The pattern of metabolic depression produced by etomidate differs markedly from that produced by barbiturates, which affect all brain regions to a similar degree. The possibility is discussed that the anesthetic effect of etomidate may be mediated by receptors.

Regional Cerebral Glucose Utilization during Althesin® Anesthesia

The effect of Althesin®, an anesthetic comprising two steroids, on regional cerebral function was determined by measurement of regional cerebral glucose utilization. Rats were anesthetized with an intravenous dose of 4, 8, or 20 mg total steroid/kg. These doses produced anesthesia for 12, 18, and 37 min, respectively. There were no physiologically significant effects of Althesin® (20 mg/kg) on body temperature, blood pH, or blood gases. Blood pressure and heart rate decreased slightly after administration of Althesin®. Althesin® had a profound effect on glucose consumption in many, but not all, cerebral structures. The forebrain (especially cerebral cortex) was affected most, while the hindbrain was much less so or not at all. This pattern of functional depression is in accord with the minimal effects observed on physiologic variables. The effects of Althesin® differ from those of other known anesthetics and suggest a unique mechanism. The possibility of action through naturally occurring steroid receptors is considered.