Effects of acetyl-L-carnitine and myo-inositol on high-energy phosphate and membrane phospholipid metabolism in zebra fish: a 31P-NMR-spectroscopy study.

Acetyl-L-carnitine (ALCAR) and myo-inositol are reported to enhance motor activity in animal models; modulate membrane phospholipid metabolism (ALCAR and myo-inositol) and high-energy phosphate metabolism (ALCAR) back to normal; and be effective treatments of major depression in humans. Fish in general and zebra fish in particular present unique animal models for the in vivo study of high-energy phosphate and membrane phospholipid metabolism by noninvasive in vivo 31P NMR. This 31P NMR study of free-swimming zebra fish showed that both ALCAR and myo-inositol decreased levels of phosphodiesters and inorganic orthophosphate and increased levels of PCr in the fish. These findings demonstrate both ALCAR and myo-inositol modulate membrane phospholipid and high-energy phosphate metabolism in free-swimming zebra fish.

Quantitative 1H and 31P MRS of PCA extracts of postmortem Alzheimer's disease brain.

Several previous studies have shown metabolic abnormalities in perchloric acid extracts of postmortem Alzheimer's disease (AD) brain by both proton (1H) and phosphorus-31 (31P) magnetic resonance spectroscopy (MRS). In all of these studies the results were expressed in relative terms, in units of mol percent. The results of this study, expressed in the absolute units of mumol/g wet weight, verify the previous 1H and 31P MRS studies. Absolute increases were found for myo-inositol, aspartate, L-glutamate, alanine, phosphocholine, and the phosphodiesters,. Absolute decreases were found for phosphoethanolamine and N-acetyl-l-aspartate. Many of these changes also were observed in non-AD dementia brain extracts, but changes in myo-inositol, inositol-l-phosphate, aspartate, and L-glutamate appeared to be more specific for AD in extracts of many brain areas. These results suggest that compounds related to membrane degradation and excitatory neuro-transmission increase in Alzheimer's disease while compounds related to neuronal integrity and inhibitory neurotransmission are decreased.

Endogenous inhibitors of human choline acetyltransferase present in Alzheimer's brain: preliminary observation.

We have previously demonstrated the presence in Alzheimer's disease brain of an endogenous inhibitor of choline acetyltransferase activity. Selected properties of these compounds were investigated. There appear to be two distinct classes of inhibitor present, both phosphomonoesters and nonphosphorylated substances. They are not proteins, pass through 500 mm dialyses membranes and are not lipoidal. There are both different sensitivities of individual control cytosotic activity to inhibition and differences in intrinsic inhibitory activity present in individual Alzheimer's disease brain samples. There is a competitive type of inhibition with respect to acetyl CoA as substrate and a noncompetitive type with respect to choline as substrate.

A preliminary 31P MRS study of autism: evidence for undersynthesis and increased degradation of brain membranes.

In this pilot study, brain high energy phosphate and membrane phospholipid metabolism were investigated in the dorsal prefrontal cortex of 11 high-functioning autistic adolescent and young adult men (the age range is 12-36 years) and 11 age-, gender-, IQ, race- and socioeconomic status-matched normal controls using in vivo 31P nuclear magnetic resonance spectroscopy (MRS). The autistic group had decreased levels of phosphocreatine and esterified ends (alpha ATP + alpha ADP + dinucleotides + diphosphosugars) compared to the controls. When the metabolite levels were compared within each subject group with neuropsychologic and language test scores, a common pattern of correlations was observed across measures in the autistic group, but not in the control group. As test performance declined in the autistic subjects, levels of the most labile high energy phosphate compound and of membrane building blocks decreased, and levels of membrane breakdown products increased. No significant correlations were present with age in either group or with IQ in the control group, suggesting that these findings were not the consequence of age or IQ effects. This pilot study provides tentative evidence of alterations in brain energy and phospholipid metabolism in autism that correlate with the neuropsychologic and language deficits. The findings are consistent with a hypermetabolic energy state and undersynthesis of brain membranes and may relate to the neurophysiologic and neuropathologic abnormalities in autism.

Alterations in brain high-energy phosphate and membrane phospholipid metabolism in first-episode, drug-naive schizophrenics. A pilot study of the dorsal prefrontal cortex by in vivo phosphorus 31 nuclear magnetic resonance spectroscopy.

In this pilot study, membrane phospholipid and high-energy phosphate metabolism were studied in the dorsal prefrontal cortex of 11 drug-naive, first-episode schizophrenic patients and compared with those of 10 healthy control volunteers comparable in age, education, and parental education. The schizophrenic patients had significantly reduced levels of phosphomonoesters and inorganic orthophosphate and significantly increased levels of phosphodiesters and adenosine triphosphate compared with the controls. The levels of phosphocreatine and adenosine diphosphate did not differ in the two subject groups. The adenosine triphosphate and inorganic orthophosphate findings suggest functional hypoactivity of the dorsal prefrontal cortex. The phosphomonoester and phosphodiester findings are compatible with either premature aging or an exaggeration of normal programmed regressive events occurring in the neural systems sampled.

Changes in brain energy and phospholipid metabolism during development and aging in the Fischer 344 rat.

The effects of brain development and aging on high-energy phosphate and membrane phospholipid metabolism were studied from birth to senescence in the Fischer 344 rat using 31P nuclear magnetic resonance spectroscopy. Marked developmental and smaller aging-related changes were observed in brain high-energy phosphates, phospholipid precursors and phospholipid breakdown products. The biochemical changes correlate with known histological and electrophysiological changes occurring in the brain during development (neuritic sprouting and onset of brain electrical activity) and aging (loss of dendritic processes). These findings provide a framework for interpreting the effects of physiological insults during different developmental and aging periods.

In vivo 31P NMR profiles of Alzheimer's disease and multiple subcortical infarct dementia.

We used in vivo phosphorus 31 nuclear magnetic resonance (31P NMR) spectroscopy to study regional high-energy phosphate and phospholipid metabolism in brains of patients with dementia associated with probable Alzheimer's disease (AD) and multiple subcortical cerebral infarctions (MSID). The MSID patients demonstrated elevations of the phosphocreatine (PCr)/inorganic orthophosphate (Pi) ratio in both the temporoparietal and frontal regions. Phosphomonoesters (PME) and the ratio of PME to phosphodiesters were elevated in the temporoparietal region of AD. Pi was also elevated in the frontal and temporoparietal regions of AD. Findings from 31P NMR were accurate in distinguishing MSID from AD. Values of PCr/Pi accurately classified 100% of the MSID patients and 92% of AD. Pi and PME, considered jointly, also accurately classified all MSID and all but 1 AD. Findings from in vivo 31P NMR spectroscopy appear to yield metabolic profiles useful in distinguishing AD from MSID.

Correlation of phosphorus-31 magnetic resonance spectroscopy and morphologic findings in Alzheimer's disease.

Senile plaques (SPs), especially, and neurofibrillary tangles are important pathologic markers for the diagnosis of Alzheimer's disease (AD), but neither is pathognomonic for AD. We hypothesize that elevations in levels of phosphomonoesters, precursors of membrane phospholipids, occur early in the pathogenesis of AD and precede the appearance of SPs. In contrast, elevations in levels of phosphodiesters, breakdown products of phospholipids, reflect degeneration of neural membranes and will correlate with the appearance of SPs. Correlative phosphorus-31 magnetic resonance spectroscopy and morphologic studies conducted to test this hypothesis disclosed that elevations in levels of phosphomonoesters had a negative correlation with the numbers of SPs, and elevations in levels of phosphodiesters had a positive correlation with the numbers of SPs. No correlations were observed for either membrane parameter and neurofibrillary tangles. These findings support our hypothesis and suggest that aberrations in the synthesis of membrane phospholipids are early metabolic events in the pathogenesis of AD.

31P nuclear magnetic resonance study of the brain in Alzheimer's disease.

The histopathological hallmarks of Alzheimer's disease have long been considered to be neurofibrillary tangles (NFT) and neuritic (senile) plaques (SP). Neither of these structures, however, are unique to Alzheimer's disease, and both probably represent end-stage markers of the disorder. NFT have been demonstrated in many disorders; SP occur in small numbers with normal aging. Evidence is presented for elevation of phosphomonoesters (PME) in Alzheimer's brain compared to non-Alzheimer's diseased controls and normal controls. The PME detected by 31P nuclear magnetic resonance (NMR) spectroscopy of autopsy brain are predominantly anabolic precursors of membrane phospholipids. Elevated PME could be secondary to a metabolic block at the rate-limiting enzyme in membrane phospholipid synthesis, which is cytidine triphosphate (CTP): phosphocholine (or phosphoethanolamine) cytidyltransferase (EC 2.7.7.15). Elevated PME could also be secondary to decreased breakdown of PME by phospholipase D activity. Since CTP: phosphocholine cytidyltransferase is inactivated by phosphorylation and since there is independent evidence for hyperphosphorylation of tau and MAP-2 proteins in AD brain, enhanced protein kinase activity could be a common factor. Preliminary evidence suggests that PME could interact with N-methyl-D-aspartate receptors and potentially act as false neurotransmitters. Further studies will be needed to investigate these possibilities.

P-31 nuclear magnetic resonance analysis of brain: normoxic and anoxic brain slices.

Perchloric acid extracts were prepared from liquid-N2-frozen gerbil and guinea pig brain slices studied under one of three conditions: O2-incubated, N2-incubated or O2-incubated recovery following N2 incubation. Mole percentages of the various phosphatic components contained in the extracts were determined by phosphorus-31 nuclear magnetic resonance spectroscopy. The brain slice extract spectrum revealed a previously unreported group of brain phosphodiesters at -0.73 delta relative to 85% orthophosphoric acid. Although the phosphatic profiles from O2-incubated slices from gerbils and guinea pigs revealed only minor species variations, which differed quantitatively rather than qualitatively, species-specific differences were made readily apparent and amplified by incubating brain slices under oxygen-deficient conditions. Despite these differences which were most prevalent during the recovery phase, the overall metabolic changes described herein in response to N2-incubation were in accord with the results obtained by other analytical techniques. Inorganic orthophosphate (2.63 delta) was increased, while nucleoside (principally, adenosine) triphosphate (alpha-, -10.92 delta, beta-, -21.45 delta, and gamma-, -5.80 delta) and phosphocreatine (-3.12 delta) levels were decreased in response to N2 incubation. In addition, inosine monophosphate (3.78 delta) was increased and the levels of a partially characterized acid-labile phosphate (0.85 delta, guinea pig) were decreased upon N2 incubation. Phosphoglyceride metabolism also appeared to be altered by oxygen deprivation (gerbil). These latter findings provide additional information concerning the metabolic responses of cerebral tissue to oxygen deficient conditions.

Phosphorus 31 NMR of neuroblastoma clonal lines. Effect of cell confluency state and dibutyryl cyclic AMP.

Phosphorus nuclear magnetic resonance is used to study changes in the levels of the major phosphate-containing intermediary metabolites concomitant with induced cellular differentiation in the N-18 and C-46 neuroblastoma clonal lines. The study reveals differences between the 31P-NMR profiles of the two clonal lines and also striking differences attendant to dibutyryl cAMP-mediated morphological differentiation in the N-18 clone. Phosphorus-31 NMR would appear to provide a new technique with which to study genetic differentiation.