Bruce S. Glaeser

Oral dyskinesia in rats following brain lesions and neuroleptic drug administration

After 10–12 weeks of chronic haloperidol administration rats with frontal cortex ablations or lesions induced by intracerebroventricular injection of 6-hydroxydopamine developed vacuous chewing behavior at a fairly stable frequency (bifrontal ablations had 15–20, 6-hydroxy-dopamine lesioned rats 7–12 chewing movements/min). This behavior persisted for 10 weeks after the last injection of haloperidol decanoate. However, rats with frontal cortex lesions developed a low rate of vacuous chewings (4–8 chewings/min) even without haloperidol administration. Bilateral intrastriatal injections of kainic acid in combination with chronic haloperidol administration did not cause chewing movements in excess of unlesioned haloperidol-treated controls.Pharmacological tests of this animal model for tardive dyskinesia (TD) revealed similarities to human TD, but also differences. Dopamine agonists (apomorphine) and antagonists (haloperidol) both lowered chewing behavior analogous to reported effects on TD and so did gabaculine. The cholinergic drugs physostigmine and pilocarpine, however, increased chewing in rats, while anticholinergics (atropine) reduced it, in contrast to reported effects on human TD.

Low cerebrospinal fluid γ‐aminobutyric acid content in seizure patients

γ-Aminobutyric acid (GABA) has been implicated in the neurochemistry of epilepsy. Lumbar cerebrospinal fluid (CSF) GABA concentrations determined using an ion-exchange fluorometric assay reflect brain GABA content. The mean lumbar CSF GABA concentration among 21 medicated patients with intractable seizures was significantly lower (p < 0.001) than that of 20 urimedicated normal volunteers. Patients with generalized tonic-clonk (grand mal) and complex partial (psychomotor) seizures had significantly lower (p < 0.05) CSF GABA concentrations than those with simple partial (focal sensory/motor) seizures. Although lumbar CSF GABA levels in our seizure patients did not significantly correlate with serum concentrations of phenytoin, phenobarbital, or primidone, additional study of medication-free epileptic patients may be required to evaluate the possibility of anticonvulsant-drug-induced CSF GABA alterations.

Changes in Brain Levels of Acidic, Basic, and Neutral Amino Acids After Consumption of Single Meals Containing Various Proportions of Protein

Abstract: Rats fasted overnight were allowed to consume single meals containing 0, 18, or 40% protein or continued to fast; after 2 h, brains and sera were taken and assayed for various amino acids. In general, serum levels of most amino acids were reduced by the 0% protein meal and elevated by the high‐protein meal when compared with those associated with fasting conditions. Exceptions were those not diminished by the 0% protein meal (tryptophan, methionine, proline) and those increased (alanine) or decreased (glycine) by all of the test meals. Amino acids exhibiting the broadest normal ranges (estimated by comparing their serum levels after 40% protein with those after 0% protein) were tyrosine, leucine, valine, isoleucine, and proline; serum lysine and histidine, two basic amino acids, also varied more than threefold. Brain levels of lysine, histidine, and some of the large neutral amino acids (LNAAs) also exhibited clear relationships to the protein content of the test meal: those of valine, leucine, and isoleucine were depressed by the 0% protein but increased (compared with 0% protein) when protein was added to the meal: brain tyrosine was increased by all of the test meals in proportion to their protein contents; tryptophan, phenylalanine, and glutamate were increased after the 0% protein meal but not by protein‐containing meals; brain lysine, histidine, and methionine were increased after the high‐protein meal, and brain alanine was increased slightly by all of the meals. For each of the LNAAs, significant correlations were observed between its brain level in any animal and the ratio of its serum concentration to the sum of the concentrations of its LNAA competitors (for blood‐brain barrier transport). For valine, tyrosine, lysine, and histidine, significant correlations were obtained between their brain and serum levels.

Effect of various oral glucose doses on plasma neutral amino acid levels

Six healthy, nonobese, fasting subjects each received, on different days 0, 6, 12.5, 25, or 50 g of glucose (Glucola) in a total volume of 100 ml. Blood was taken at intervals and assayed for plasma levels of the branched-chain amino acids (valine, isoleucine and leucine); the other major large neutral amino acids (LNAA) (methionine, phenylalanine, tyrosine and tryptophan); and, in some cases, insulin and glucose. Insulin levels were significantly elevated 30 min after consumption of 12.5, 25, or 50 g of glucose, and were higher after the 50 g dose than after 12.5 g. Changes in plasma glucose concentrations were small and did not correlate with glucose dose. Mean percent reductions of LNAA tended to exhibit dose-dependence, most clearly observed after 120 min. In some subjects as little as 6 g of glucose transiently decreased LNAA concentrations. Branched-chain amino acids were most sensitive, decreasing by 35%-41% after 50 g of glucose. Plasma tryptophan concentrations fell only by 23%, hence the ratio of plasma tryptophan to other plasma LNAA (which affects brain serotonin synthesis) increased significantly.

Elevation of plasma tyrosine after a single oral dose of L-tyrosine

Abstract Plasma tyrosine concentrations in twelve normal, fasting human subjects were significantly elevated 2–8 hours after they ingested 100 mg/kg or 150 mg/kg tyrosine. Mean plasma tyrosine levels were maximal after 2 hours, rising from 69 ± 3.9 to 154 ± 9.5 nmols/ml ( X ± SEM ) after the 100 mg/kg dose and to 203 ± 31.5 nmols/ml after the 150 mg/kg dose (p ≤ 0.001 for both doses). The mean tyrosine ratio (defined as the ratio of plasma tyrosine concentration to the sum of the concentrations of six other neutral amino acids that compete for the same blood-brain barrier uptake system) increased from 0.10 ± 0.02 to 0.28 ± 0.04 ( X ± SEM ) 2 hours after the 100 mg/kg dose (p ≤ 0.001) and to 0.35 ± 0.05 2 hours after the 150 mg/kg dose (p ≤ 0.005). No side effects of orally-administered L-tyrosine were noted.

Plasma tyrosine in normal humans: Effects of oral tyrosine and protein-containing meals

To test the effects of tyrosine ingestion and concurrent food consumption on plasma tyrosine levels and on the plasma tyrosine ratio, we measured plasma neutral amino acid levels in 11 subjects who consumed a diet containing 113 g protein and who also took 100 mg/kg/day of L-tyrosine (in three equally divided doses) before meals. Plasma tyrosine levels rose significantly (p<0.025) during the day when subjects consumed the diet alone; they increased markedly after tyrosine ingestion (p<0.005). Tyrosine administration did not affect plasma concentrations of the other neutral amino acids that compete with tyrosine for entry into the brain. Thus, the plasma tyrosine ratio increased from 0.13 to 0.21 (p<0.001) on the day fed subjects received the tyrosine. These observations indicate that tyrosine administration might increase brain tyrosine levels and perhaps accelerate catecholamine synthesis in humans with diseases in which catecholamine synthesis or release is deficient.

Recombinant human ciliary neurotrophic factor stimulates the metabolic activity of SH-SY5Y cells as measured by a cytosensor microphysiometer

Information on the transmembrane signaling events and subsequent biochemical processes initiated by ciliary neurotrophic factor (CNTF) receptor activation in neurons is lacking. SH-SY5Y cells, a human neuroblastoma cell line expressing CNTF receptors, were used to study metabolic changes associated with functional ligand-receptor interactions. Real-time measurements quantifying the rate of extracellular acidification by SH-SY5Y cells (a measure of metabolic activity) were made using a silicon-based cytosensor. Application of recombinant human CNTF (rhCNTF) to resting SH-SY5Y cells increased their acidification rate in a concentration and time-dependent manner with an apparent EC50 of 60 ng/ml. Pretreatment of cells with phosphatidylinositol-specific phospholipase C (PI-PLC) prevented the CNTF, but not an NGF-stimulated increase in acidification rate. Collectively, these results demonstrate that: (1) SH-SY5Y cells express functional CNTF receptors; and (2) the initial signal transduction mechanism activated by the CNTF receptor in SH-SY5Y cells is distinct from that activated by the NGF receptor; however, both may ultimately stimulate the same downstream biochemical messengers to increase cellular metabolism.