B. Ghigini

Acetylcholinesterase activity of synaptic plasma membranes during ageing: effect of L-acetylcarnitine.

A physiopathological role for acetylcholine (ACh) was hypothesized during ageing and related neurodegenerative diseases, e.g. dementia. This research was aimed to study acetylcholinesterase (AChE) activity during development and ageing of the frontal cerebral cortex of 4-, 8-, 12-, 16-, 20- and 24-month-old rats. This study was performed on synaptic plasma membranes, the specific subcellular compartment where the enzyme is located in vivo both in control animals and after in vivo acute treatment with L-acetylcarnitine. Maximum AChE activity was unaffected by age, and L-acetylcarnitine treatment increased enzyme activity in synaptic plasma membranes of 8-month-old rats. A comprehensive analysis of these results suggests: (a) the observed alterations in protein can substantially affect neurochemical data if results are presented as specific activities per unit protein; (b) energy metabolism plays the major role in the disturbed ACh metabolism during ageing and (c) the understanding of the mode of action of L-acetylcarnitine in treatment of dementia.

Mitochondrial factors involved in Parkinson's disease by MPTP toxicity inMacaca fascicularis and drug effect

The maximal rates (Vmax) of some mitochondrial enzyme activities related to energy transduction (citrate synthase, succinate dehydrogenase, malate dehydrogenase, NADH-cytochrome c reductase, cytochrome oxidase) and amino acid metabolism (glutamate dehydrogenase, glutamate-pyruvate-and glutamate-oxaloacetate-transaminases) were evaluated in non-synaptic (“free”) and intrasynaptic “light” and “heavy” mitochondria fromhippocampus ofMacaca fascicularis (Cynomolgus monkey). The different mitochondrial populations were isolated from thehippocampus of monkeys treated p.o. with dihydroergocriptine at a dose of 12 mg/kg/day before and during the induction of a Parkinsons-like syndrome by MPTP administration (i.v., 0.3 mg/kg/day for 5 days). The MPTP administration modified the activity of some enzymes related to the metabolism of glutamate and the activity of succinate dehydrogenase on selected types of mitochondria. Pharmacological treatment by dihydroergocriptine promoted return to the steady-state levels of most enzymes, demonstrating a protective effect on these biochemical parameters.

Age-related alterations by chronic intermittent hypoxia on cerebral synaptosomal ATPase activities.

The age-related alterations in the plasticity of synaptic energy-requiring ATPases [Na+,K(+)-ATPase, low- and high-affinity Ca(2+)-ATPase, Mg(2+)-ATPase, and Ca2+,Mg(2+)-ATPase] were assayed in synaptosomes and synaptosomal subfractions [namely, synaptosomal plasma membranes and synaptic vesicles] in the cerebral cortex from 3- and 24-month-old normoxic rats and rats subjected to either mild or severe chronic (four weeks) intermittent normobaric hypoxia. With the exception of the high-affinity Ca(2+)-ATPase, aging induced a decrease in the ATPase activities from normoxic rats. The adaptation to mild hypoxia was characterized by an increase in the activity of Mg(2+)-ATPase in 3-month-old rats, concomitant with a decrease in the activities of: (i) Na+,K(+)-ATPase and high-affinity Ca(2+)-ATPase in both 3- and 24-month-old rats, and (ii) Ca2+,Mg(2+)-ATPase in 3-month-old ones. The adaptation to chronic intermittent severe hypoxia was characterized by a decrease in the activities of: (i) Na+,K(+)-ATPase, Ca2+,Mg(2+)-ATPase and high-affinity Ca(2+)-ATPase in both 3- and 24-month-old rats, and (ii) low-affinity Ca(2+)-ATPase only in 24-month-old ones. The effect on Mg(2+)-ATPase activity was characterized by a decrease in the activity of the enzymatic form located in the synaptic plasma membranes [involved in ATP hydrolysis to adenosine production], concomitant with an increase in the activity of the form located in the synaptic vesicles [involved in the turnover of transmitters, e.g., glutamate].

Modifications by hypoxia and drug treatment of cerebral ATPase plasticity.

The plasticity of synaptosomal non-mitochondrial ATPases was evaluated in cerebral cortex from 3-month-old normoxic rats and rats subjected to either mild or severe intermittent normobaric hypoxia [12 hr daily exposure to N2∶O2 (90∶10 or 91.5∶8.5) for four weeks]. The activities of Na+, K+-ATPase, low- and high-affinity Ca2+-ATPase, Mg2+-ATPase, and Ca2+, Mg2+-ATPase were assayed in synaptosomes and synaptosomal subfractions, namely synaptosomal plasma membranes and synaptic vesicles. The evaluations were performed after a 4-week treatment with saline (controls) or α-adrenergic agents (δ-yohimbine, clonidine), a vasodilator compound (papaverine), and an oxygen-partial pressure increasing agent (almitrine). These treatments differently changed the adaptation to chronic intermittent hypoxia characterized by a decrease in the activity of Na+, K+-ATPase, Ca2+,Mg2+-ATPase, and high-affinity Ca2+-ATPase, concomitant with a modification in the activity of Mg2+-ATPase supported in a different way by the enzymatic forms located into the synaptosomal plasma membranes and synaptic vesicles.

Synaptosomal non-mitochondrial ATPase activities and drug treatment

Energy-using non-mitochondrial ATPases were assayed in rat cerebral cortex synaptosomes and synaptosomal subfractions, namely synaptosomal plasma membranes and synaptic vesicles. The following enzyme activities were evaluated: Na+, K+-ATPase; high- and low-affinity Ca2+-ATPase; basal Mg2+-ATPase; Ca2+, Mg2+-ATPase. The evaluations were performed after four week-treatment with saline [controls] or α-adrenergic agents (δ-yohimbine, clonidine), energymetabolism interfering compound (theniloxazine), and oxygen-partial pressure increasing agent (almitrine), in order to define the plasticity and the selective changes in individual ATPases. In rat cerebral cortex, the enzyme adaptation to four-week-treatment with δ-yohimbine or clonidine was characterized by increase in both high- and low-affinity Ca2+-ATPase activities. The action involves the enzyme form located in the synaptic plasma membranes. The enzyme adaptation to the subchronic treatments with theniloxazine or almitrine was characterized by increase in Na+, K+-ATPase or Mg2+-ATPase activities, respectively. The action involves the enzymatic forms located in the synaptic plasma membranes. Thus, the pharmacodynamic effects of the agents tested should also be related to the changes induced in the activity of some specific synaptosomal nonmitochondrial ATPases.

Synaptosomal non-mitochondrial ATPase activities: Age-related alterations by chronic normobaric intermittent hypoxia

In synaptosomes and synaptosomal subfractions (namely, synaptosomal plasma membranes and synaptic vesicles) the age-related alteration in the plasticity of synaptic energy-requiring ATPases (Na+, K(+)-ATPase, low- and high-affinity Ca(2+)-ATPase, Mg(2+)-ATPase and Ca2+, Mg(2+)-ATPase) were assayed in the cerebral cortex from 3- and 24-month-old normoxic rats and rats subjected to either mild or severe chronic (4 weeks) intermittent normobaric hypoxia. With the exception of the high-affinity Ca(2+)-ATPase, aging induced a decrease in the ATPase activities from normoxic rats. The adaptation to mild hypoxia was characterized by an increase in the activity of Mg(2+)-ATPase in 3-month-old rats, concomitant with a decrease in the activities of: (i) Na+,K(+)-ATPase and high-affinity Ca(2+)-ATPase in both 3- and 24-month-old rats; and (ii) Ca2+,Mg(2+)-ATPase in 3-month-old ones. The adaptation to chronic intermittent severe hypoxia was characterized by a decrease in the activities of: (i) Na+,K(+)-ATPase, Ca2+,Mg(2+)-ATPase and high-affinity Ca(2+)-ATPase in both 3- and 24-month-old rats and (ii) low-affinity Ca(2+)-ATPase only in 24-month-old ones. The effect on Mg(2+)-ATPase activity was characterized by a decrease in the activity of the enzymatic form located in the synaptic plasma membranes (involved in ATP hydrolysis to adenosine production), concomitant with an increase in the activity of the form located in the synaptic vesicles (involved in the turnover of transmitters, e.g., glutamate).

Age-related changes by hypoxia and trh analogue on synaptic atpase activities

Some synaptosomal energy-requiring ATPases were evaluated in the cerebral cortex from 3- and 24-month-old normoxic rats and rats submitted to either mild or severe chronic (4 weeks) intermittent normobaric hypoxia. Furthermore, 4-week treatment with saline or TRH analogue posatireline was performed. The activities of Na+,K(+)-ATPase, low- and high-affinity Ca(2+)-ATPase, and Ca2+,Mg(2+)-ATPase were assayed in synaptosomes and synaptosomal subfractions, namely synaptosomal plasma membranes and synaptic vesicles. With the exception of the high-affinity Ca(2+)-ATPase, aging induced a decrease in the ATPase activities from normoxic rats. The adaptation to chronic intermittent mild hypoxia was characterized by an increase in the activity of Mg(2+)-ATPase in 3-month-old rats, concomitant with a decrease in the activities of: a) Na+,K(+)-ATPase and high-affinity Ca(2+)-ATPase in both 3- and 24-month-old rats, and b) Ca2+,Mg(2+)-ATPase in 3-month-old ones. The TRH analogue posatireline increased the high-affinity Ca(2+)-ATPase in both 3- and 24-month-old hypoxic rats, concomitant with an increase in Mg(2+)-ATPase activity in 24-month-old ones. The adaptation to chronic intermittent severe hypoxia was characterized by a decrease in the activities of: a) Na+,K(+)-ATPase, Ca2+,Mg(2+)-ATPase and high-affinity Ca(2+)-ATPase in both 3- and 24-month-old rats, and b) low-affinity Ca(2+)-ATPase only in 24-month-old ones. The effect on Mg(2+)-ATPase activity was characterized by a decrease in the enzymatic form located in the synaptic plasma membranes, concomitant with an increase in the form located in the synaptic vesicles.(ABSTRACT TRUNCATED AT 250 WORDS)

Is the Mg2+-ATP-dependent proton pumping activity of the synaptic vesicles a factor involved in the cerebral hypoxia ?

AbstractThe changes in the Mg2+-dependent V-type ATPase activity and the Mg2+-ATP-dependent H+ pumping activity of the synaptic vesicles from the cerebral cortex of rats submitted to intermittent chronic (4 weeks) mild or severe hypoxia were evaluated. The adaptation to the chronic severe hypoxia increases both the ATPase and the H+ pumping activities which are inhibited by NEM with an exponential relationship between the IC50 values and the in vivo O2 concentration. The Mg2+-dependent increase in H+ pumping activity of synaptic vesicles from the rats subjected to in vivo chronic hypoxia may be antagonized by nigericin (dissipating ΔpH) and by FCCP (dissipating ΔpH and ΔΨSV). In contrast, valinomycin (dissipating the ΔΨSV and facilitating an enhancement in ΔpH) increases in vitro the H+ pumping activity that is inhibited by the addition of high concentration of K gluconate (reducing the rate of K+ efflux). The preincubation of vesicles from hypoxic rats with FCCP, but not with nigericin, inhibits the valinomycin-increased H+ pumping activity.l-glutamate increases the H+ pumping activity in synaptic vesicles from the cerebral cortex of chronic hypoxic rats, whereas other amino acids (i.e.,l-aspartate andl-homocysteate) and glutamate analogs (i.e., quisqualate and ibotenate) are ineffective. The adaptation to both chronic intermittent severe hypoxia and in vivo treatment with posatireline causes a decrease in the Mg2+-ATPase activity consistent with the decrease in the H+ pumping one of the synaptic vesicles. The addition of nigericin into incubation medium magnifies the decrease in the H+ pumping activity, while the addition of FCCP is ineffective, suggesting that the treatment with posatireline interferes with the ΔΨSV component in the

Effect of intermittent mild hypoxia and drug treatment on synaptosomal nonmitochondrial ATPase activities.

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