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J Bothmer

Publications and source records attributed to J Bothmer.

10 recordsLinked to original sources
PubMed1994

Phosphoinositide kinase activities in synaptosomes prepared from brains of patients with Alzheimer's disease and controls.

Previously, phosphatidylinositol (PI) kinase activity in cytosolic fractions prepared from postmortem tissue of the cerebral cortex from patients with Alzheimer's disease (AD) appeared to be lower than that of age-matched controls [Jolles et al., J. Neurochem., 58 (1992) 2326-2329]. In the study presented here, PI and PIP (phosphatidylinositol phosphate) kinase activities were studied in synaptosomes prepared from postmortem brain tissue of AD patients and age-matched controls. Firstly, PI kinase activity in synaptosomes prepared from the frontal superior gyrus of AD brain was 30% lower than in synaptosomes prepared from postmortem tissue of control brain. PIP kinase activity was the same in AD and control synaptosomes. Secondly, the yield of synaptosomal protein (micrograms protein per mg tissue wet weight) was lower in preparations from AD brain than in preparations from control brain, which could be a manifestation of a loss of presynaptic terminals in the frontal cortex. These results suggest that the difference in PI kinase activity between AD and control brain tissue may originate from differences in intact neurons in view of the fact that synaptosomes can originate only from intact neurons.

1-Phosphatidylinositol 4-Kinase↗
PubMed1994

The effect of age on phosphatidylinositol kinase, phosphatidylinositol phosphate kinase and diacylglycerol kinase activities in rat brain cortex.

A previous study, in which a lysed fraction was used with endogenous phospholipids as substrate, revealed age-related changes in PA and PIP2 formation but not in PIP formation (Bothmer et al., Neurochem. Int. 21, 223-228, 1992). To rule out the influence of substrate availability in the present study, the effect of age on PI kinase, PIP kinase and DAG kinase activities was studied with exogenous phospholipids as substrate in the cerebral cortex from 8-month-old, 14-month-old and 26-month-old Brown Norway rats. PI kinase activity was predominantly located in a tight membrane-bound protein fraction, DAG kinase activity in cytosolic and loosely membrane-bound protein fractions, and PIP kinase activity was present in all three protein preparations. The effects of age were limited to a small increase in kinase activity in the tight membrane-bound protein fraction in 14-month-old and 26-month-old rats compared to 8-month-old rats, and a 10% decrease in PIP kinase activity in the cytosolic protein fraction in 14-month-old and 26-month-old rats compared to 8-month-old rats. DAG kinase activity showed no age-related changes. In conclusion, one should take care in comparing rat aging with human aging as PI kinase activity shows an age-related decline in human brain cortex (Jolles et al., J. Neurochem. 58, 2326-2329, 1992). Furthermore, previously reported decreases in PA formation rates in rat brain are probably not due to changes in DAG kinase itself but to changes in DAG availability, although further experimental evidence is needed to confirm this conclusion.

1-Phosphatidylinositol 4-Kinase↗
PubMed1993

Behavioural and biochemical effects of acute central metabolic inhibition: effects of acetyl-l-carnitine.

In the present study we evaluated a new method to assess the behavioural and biochemical effects of a brief period of acute hypoxia in the brain. In this method, cyanide is injected into the lateral ventricles. Spatial navigation performance in a Morris task was found to be impaired 1 and 5 min after an i.c.v. injection of 5.0 micrograms cyanide but not after 2.5 micrograms cyanide. Increased rate of phosphatidic acid formation, reflecting increased phospholipase C activity, were observed after injection of 5.0 micrograms cyanide, indicating that energy-dependent phosphoinositide metabolism was affected. Chronic treatment with acetyl-l-carnitine attenuated the cyanide-induced behavioural deficit, but had no effect on energy-dependent phosphoinositide metabolism. The results suggest that, in this model, acetyl-l-carnitine may act via free fatty acid metabolism, by increasing the reservoir of activated acyl groups which are involved in the reacylation of membrane phospholipids.

Acetylcarnitine↗
PubMed1993

Platelet phosphatidylinositol kinase activity is not altered in Alzheimer disease.

We previously reported a specific decline in phosphatidylinositol (PI) kinase activity in the neocortex of patients with Alzheimer disease (AD) as compared to controls, whereas phosphatidylinositol phosphate (PIP) kinase activity appeared not to be affected (Jolles et al., 1992). In search of a possible systemic effect of AD, in the present study we investigated phosphoinositide kinase activity in platelets from patients with AD and from control subjects. The study was based on the notion that disease-specific abnormalities in the brain could be reflected in blood platelets. PI kinase activity was studied in platelet homogenates and in a salt-solubilized protein fraction of platelets, because of the difference in subcellular localization of the different types of PI kinases. In addition, NADH cytochrome-C reductase was measured in platelet homogenates as a marker for the endoplasmic reticulum, to detect a possible proliferation of the endoplasmic reticulum. AD patients and normal elderly controls showed no difference in PI kinase activity in either enzyme fraction. Furthermore, NADH cytochrome-C reductase activity and the protein/phospholipid ratio per 10(6) platelets were the same for AD patients and controls. This was taken as an indication that platelets in AD patients do not show proliferation of intracellular membranes.

1-Phosphatidylinositol 4-Kinase↗
PubMed1992

Evidence for a new inositol phospholipid in rat brain mitochondria.

Phosphorylation of phosphatidylinositol (PI), phosphatidylinositol monophosphate (PIP) and diacylglycerol (DAG) was studied in rat brain cortex myelin, synaptosomal and mitochondrial fractions, with ATP as phosphate donor and endogenous phospholipids as substrate. All fractions had PI, PIP and DAG phosphorylating activity with their own characteristic subcellular distribution. However, in the mitochondrial fraction an unidentified lipid was phosphorylated, which had a slower Rf value than PIP2 on TLC. After hydrolysis of the polar head group of the lipid and separation on anion exchange columns, it appeared to be a phosphoinositide. The elution profile showed that it was not phosphatidylinositol trisphosphate, or a lyso-compound. The available evidence suggests that the unknown inositol phospholipid in rat brain mitochondria is a phosphatidylinositol 4,5-bisphosphate isomer, although the possibility of it being a glycosyl-phosphoinositide cannot be excluded.

Adenosine Triphosphate↗
PubMed1992

Brain phosphatidic acid and polyphosphoinositide formation in a broken cell preparation: regional distribution and the effect of age.

The effect of age on phosphate incorporation into phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol 4,5-bisphosphate (PIP2) and phosphatidic acid (PA) was studied. Lysed crude synaptosomal fractions of different brain regions of 3-month-old and 32-month-old Brown Norway rats were used. The brain regions tested were the hippocampus, frontal cortex, occipital/parietal cortex, entorhinal/pyriformal cortex, striatum/septum, thalamus and hypothalamus. The individual specific phosphorylating activities were unevenly distributed within the brain of Brown Norway rats. Strikingly, the distribution of phosphate incorporation into PIP2 was opposite from that of phosphate incorporation into PA. Phosphate incorporation into PA decreased (-15%) with age in almost all brain regions tested, whereas phosphate incorporation into PIP2 decreased with age only in the frontal cortex (-20%) and in the hypothalamus (-8%). The effects of age may reflect a deterioration of phosphoinositide metabolism, with its function in signal transduction coupled to receptors via G-proteins, in the brain regions involved. In addition, there was an age related decrease in protein content and total phospholipid phosphorus content of lysed crude synaptosomal preparations of all brain regions. The high correlation between the changes in these parameters may be indicative of a decrease in the number or size of synaptosomes with age in the brain regions involved.

Adenosine Triphosphate↗
PubMed1992

Phosphatidylinositol kinase is reduced in Alzheimer's disease.

Phosphatidylinositol (PI) kinase and PI phosphate (PIP) kinase activities were measured in postmortem samples of brain tissue from patients with Alzheimer's disease and nondemented control subjects. A membrane-free cytosolic fraction from four neocortical locations, with exogenous inositol lipids as the substrate, was used. Tissue from patients with Alzheimer's disease was characterized by reduced PIP formation; the reduction was 50% in prefrontal cortex, temporal cortex, and parietal cortex and 40% in precentral gyrus. In contrast, no alterations were found in PI bisphosphate formation in these four neocortical locations. The specific changes in PI kinase but not PIP kinase activity suggest that the findings may have functional relevance to the involvement of brain membrane processes in Alzheimer's disease.

1-Phosphatidylinositol 4-Kinase↗
PubMedDate not supplied

Evidence for a selective decrease in type 1 phosphatidylinositol kinase activity in brains of patients with Alzheimer's disease.

We have previously shown that phosphatidylinositol (PI) kinase activity is 40-50% lower in cytosolic fractions of neocortical regions from brains of patients with Alzheimer's disease (AD) in contrast to phosphatidylinositol phosphate (PIP) kinase activity, which was not affected. After preparing different enzyme fractions by solubilization, the PI kinase activity in the salt-solubilized protein preparation of the temporal cortex of AD brains was predominantly affected (70% decrease). PIP kinase activity in AD brains was not different from that of control brains in any of the fractions tested. PI kinase in the salt-solubilized fractions was inhibited (-75%) by 1% Triton X-100, whereas the PI kinase in the detergent solubilized protein preparation was stimulated (+80%) by 1% Triton X-100. PI kinase activity in the salt-solubilized protein preparation was almost unaffected by adenosine in contrast to PI kinase activity in the detergent solubilized protein preparation, which was strongly inhibited by adenosine. These results indicate that the PI kinase that is specifically affected in AD is the PI 3-kinase, or type 1 PI kinase, because the fraction which was affected most severely has (1) a cytosolic localization; (2) a high sensitivity to inhibition by the non-ionic detergent Triton X-100, and (3) an insensitivity to adenosine inhibition, which are characteristic features of type 1 PI kinase. The relevance of our findings is that type 1 PI kinase is thought to be involved in the regulation of cytoskeletal turnover processes.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Phosphatidylinositol 4-Kinase↗
PubMedDate not supplied

Reduced phosphatidylinositol kinase activity in Alzheimer's disease: effects of age and onset.

Phosphatidylinositol kinase (PI kinase) and phosphatidylinositol phosphate kinase (PIP kinase) were assayed in a membrane-free cytosolic fraction prepared from the medial temporal cortex of patients with Alzheimer's disease (AD) and nondemented controls, with exogenous lipids as substrate. 32P-PIP formation appeared to be reduced by 21% in patients with AD who died before 80 years, compared to matched controls. In addition, there was an age-related decrease in PI kinase activity in the control group. In the AD patients there was no age-related decrease. Very old AD patients (older than 80 when they died) with a short duration of the disease were characterized by increased PI kinase activity. No differences were found in the closely related enzyme PIP kinase. The results are indicative for heterogeneity in AD.

1-Phosphatidylinositol 4-Kinase↗