Gary M Fiskum

Publication Detail

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Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation.

Tibor Kristián; Irene B Hopkins; Mary C McKenna; Gary Fiskum (Profiled Authors: Gary M Fiskum; Mary C McKenna)

Department of Anesthesiology, School of Medicine, University of Maryland, 685 West Baltimore Street, MSTF 534, Baltimore, MD 21201, USA. tkristian@anesthlab.umm.edu
Journal of neuroscience methods 2006;152(1-2):136-43.

To study neurons or glia-specific mitochondria one needs to isolate these organelles from primary neuronal or astrocytic cell culture. This work provides novel method for isolation of functional and morphologically intact mitochondria from neurons and astrocytes in cell cultures. In the first step, mitochondria are released from cells by disruption of cell membranes using a nitrogen cavitation technique. This technique is based on rapid decompression of a cell suspension from within a pressure vessel. Mitochondria released from cell bodies are then separated from the rest of cell homogenate by Percoll gradient centrifugation. This is a relatively rapid technique that yields to very well coupled mitochondria that exhibited functional and morphological characteristics comparable to mitochondria isolated from brain tissue using common techniques. This technique thus will allow examination of mitochondria that are exclusively cell specific in origin.

4 Originating Grant

• 1.

  Kristian, Tibor

  Separation of brain glial and neuronal mitochondria

  15 July 2005 - 30 April 2008

  NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE

• 2.

  MCKENNA, MARY C

  Metabolic &Developmental Aspects of Intellectual Disability

  1 May 1997 - 31 January 2014

  EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH &HUMAN DEVELOPMENT

• 3.

  Zielke, H Ronald

  Metabolic & Developmental Aspects of Mental Retardation

  1 May 1997 - 31 January 2009

  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT

• 4.

  Fiskum, Gary

  Molecular Mechanisms of Ischemia Reperfusion Brain Injury

  1 May 1995 - 31 December 2008

  NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE

Scientific Context

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