Isolation of brain mitochondria

Løvtrup, S; Zelander, T.

doi:10.1016/0014-4827(62)90012-5

Cited by 70 publications

(12 citation statements)

References 4 publications

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“…Two peaks of ME activity were eluted from the column at concentrations of magnesium acetate of 8-13 and 30-50 mM, respectively. Mitochondria were isolated from bovine brain according to the procedure of Lovtrup and Zelander (1962), and ME activity obtained from these organelles was eluted from the DE column in a single peak at magnesium acetate concentrations of 30-50 mA4 (data not shown). Therefore, in the preparation from whole brain, the peak eluting from the DE column at low magnesium acetate concentrations was considered to be cME and purified further.…”

Section: Resultsmentioning

confidence: 99%

Purification of Cytosolic Malic Enzyme from Bovine Brain, Generation of Monoclonal Antibodies, and Immunocytochemical Localization of the Enzyme in Glial Cells of Neural Primary Cultures

Kurz

Wiesinger

Hamprecht

1993

Journal of Neurochemistry

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Cytosolic malic enzyme (EC 1.1.1.40) was purified from bovine brain 5,600-fold to a specific activity of 47 U/mg. The enzyme is a homotetramer with a subunit molecular mass of 60 kDa and an isoelectric point of 6.2. Mouse monoclonal antibodies raised against this enzyme were purified and shown to be monospecific, as indicated by immunoblotting. Immunocytochemical examination of rat astroglia-rich primary cultures at the light microscopic level revealed colocalization of cytosolic malic enzyme with the astroglial marker glial fibrillary acidic protein. Also, a colocalization with the oligodendroglial marker myelin basic protein was found. Neurons in rat neuron-rich primary cultures did not show positive staining. The data suggest that cytosolic malic enzyme is a glial enzyme and is lacking in neurons.

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Section: Resultsmentioning

confidence: 99%

Purification of Cytosolic Malic Enzyme from Bovine Brain, Generation of Monoclonal Antibodies, and Immunocytochemical Localization of the Enzyme in Glial Cells of Neural Primary Cultures

Kurz

Wiesinger

Hamprecht

1993

Journal of Neurochemistry

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“…Enzynw uL~.suj: At postnatal day 38, another set of five rats from each group were sacrificed and brains were rapidly removed, homogenized and subjected to subcellular fractionation according to the method of Lovtrup & Zelander (1962). Monoamine oxidase (MAO) was assayed in mitochondria1 fraction according to the method of Tabor rf ul.…”

Section: Netrroclienricul Estimations Polyaniine Estimutionmentioning

confidence: 99%

Effect of Deltamethrin on Regional Brain Polyamines and Behaviour in Young Rats

Malaviya

Seth

Husain

1994

Pharmacology & Toxicology

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The present study examines the mechanism of neurotoxic action of a synthetic pyrethroid formulation deltamethrin in young rats. Newly weaned Wistar Albino male rats received deltamethrin of technical grade at a dose of 7.0 mg/kg body weight/day in corn oil, orally from postnatal day 22 to postnatal day 37. Deltamethrin significantly decreased the wet weight of the hippocampus without much affecting the weight of cerebellum, pons medulla, hypothalamus, frontal cortex and corpus striatum in comparison to respective controls. A significant increase in the activities of mitochondrial monoamine oxidase and microsomal acetylcholinesterase without any effect on microsomal Na+, K(+)-ATPase activity was observed in the brain of experimental animals. Our results further indicate that deltamethrin markedly impaired learning function and significantly increased the spontaneous locomotor activity while aggressive behaviour remained unaffected. An overall enhancement of polyamine levels in hypothalamus and corpus striatum accompanied with an overall decline in pon medulla and cerebellum was also noted. Maximum decrease of spermine and spermidine was registered in hippocampus, while these polyamines showed an increase in frontal cortex. In striatal membranes the binding of 3H-spiperone decreased and 3H-quinuclidinyl benzilate was elevated significantly. Deltamethrin-induced deviations in regional brain polyamine levels may be a possible cause for altered pathophysiology of the neurone.

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“…Cells were detached with a rubber policeman and homogenized with a Potter-Elvehjem homogenizer. Mitochondria were isolated by differential centrifugation as described (Lovtrup and Zelander, 1962). They were disrupted by incubation for 1 h in 20 mM Tris/ HC1, 0.1 % (mass/vol.)…”

Section: Methodsmentioning

confidence: 99%

Significant Amounts of Glycogen are Synthesized from 3‐Carbon Compounds in Astroglial Primary Cultures from Mice with Participation of the Mitochondrial Phosphoenolpyruvate Carboxykinase Isoenzyme

Schmoll

Führmann

Gebhardt

et al. 1995

European Journal of Biochemistry

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The incorporation was studied of the gluconeogenic substrates lactate, alanine, aspartate and glutamate into glycogen of astroglial primary cultures derived from mouse brain. The incorporation was inhibited by 3-mercaptopicolinate, an inhibitor of one of the characteristic gluconeogenic enzymes, phosphoenolpyruvate carboxykinase. Only the mitochondrial isoenzyme of phosphoenolpyruvate carboxykinase was detect,able in the astroglial primary cultures. After the incubation of glucose-starved cells with medium containing a mixture of [6-3H]glucose and [U-'4C]glucose, the newly synthesized glycogen showed a 3W 14C ratio which was approximately 15% less than the isotope ratio for the medium. The decrease of the isotope ratio was not significantly inhibited by 3-mercaptopicolinate, indicating a cycling of approximately 15% of the glucose to the level of the triose phosphates before its incorporation into astroglial glycogen. During the initial phase of glycogen resynthesis, the contribution of the gluconeogenic substrates appeared to be higher. This was in agreement with the accumulation of fructose 2,6-bisphosphate during refeeding. A participation of gluconeogenic substrates in glycogen metabolism was also detectable when the glycogen content was not changing significantly.

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Isolation of brain mitochondria

Cited by 70 publications

References 4 publications

Purification of Cytosolic Malic Enzyme from Bovine Brain, Generation of Monoclonal Antibodies, and Immunocytochemical Localization of the Enzyme in Glial Cells of Neural Primary Cultures

Purification of Cytosolic Malic Enzyme from Bovine Brain, Generation of Monoclonal Antibodies, and Immunocytochemical Localization of the Enzyme in Glial Cells of Neural Primary Cultures

Effect of Deltamethrin on Regional Brain Polyamines and Behaviour in Young Rats

Significant Amounts of Glycogen are Synthesized from 3‐Carbon Compounds in Astroglial Primary Cultures from Mice with Participation of the Mitochondrial Phosphoenolpyruvate Carboxykinase Isoenzyme

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