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Aldolase C/Zebrin II is Released to the Extracellular Space after Stroke and Inhibits the Network Activity of Cortical Neurons

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Abstract

Cell death after stroke involves apoptotic, autophagocytic and necrotic mechanisms which may cause the release of cytosolic proteins to the extracellular space. Aldolase C (AldC) is the brain specific isoform of the glycolytic enzyme fructose-1,6-bisphosphate aldolase. According to its characteristic striped expression pattern in the adult cerebellum AldC is also termed zebrin II. Here, we demonstrate release of AldC into the cerebrospinal fluid (CSF) after stroke in vivo. Studies with cell cultures confirmed that AldC is released to the extracellular space after hypoxia. Moreover, addition of purified recombinant AldC to networks of cortical neurons plated on multielectrode arrays reversibly inhibited the spontaneous generation of action potentials at AldC concentrations which can be expected to occur after lesions of the human cerebral cortex. This mechanism could be relevant in the pathogenesis of the electrophysiological changes in the penumbra region after stroke.

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Abbreviations

ABTS:

2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)

A.d.:

Aqua dest (H2O)

AldA:

Aldolase A

AldB:

Aldolase B

AldC:

Aldolase C

BSA:

Bovine serum albumin

CSF:

Cerebrospinal fluid

DTT:

Dithiothreitol

ECL:

Enhanced chemiluminescence

FBP:

Fructose-1,6-bisphosphate

GAP:

Glycerinaldehyde-3-phosphate

GDH:

Glycerol-3-phosphate-dehydrogenase

IPTG:

Isopropyl-β-d-thiogalactopyranosid

kDa:

kilo Dalton

LB:

Luria Bertani

MRI:

Magnetic resonance imaging

PBS:

Phosphate buffered saline

PMSF:

Phenyl methyl sulfonyl fluoride

RT:

Room temperature

SDS-PAGE:

Sodium dodecylsulfate-polyacrylamide gel electrophoresis

TBS:

Tris buffered saline

TIM:

Triose-phosphate-isomerase

TTX:

Tetrodotoxin

WB:

Western blot

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Acknowledgments

We thank Norbert Rösel for excellent technical assistance and Prof. Richard Hawkes for the gift of the zebrin II antibody. This study was supported in part by the BMFZ of the Heinrich-Heine-University (Germany) and the Pinguin-Stiftung (Düsseldorf/Germany).

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Author notes

  1. Stephan L. Baader

    Present address: Institute of Anatomy I, University of Jena, Teichgraben 7, Jena, D-07743, Germany

  2. Ulrich Junghans

    Present address: Sana-Klinikum Remscheid, Burger Straße 211, D-42859, Remscheid, Germany

Authors and Affiliations

  1. Institut für Physiologische Chemie, Rheinische Friedrich-Wilhelms-Universität Bonn, Nussallee 11, D-53115, Bonn, Germany

    Stephanie Linke, Volkmar Gieselmann & Joachim Kappler

  2. Neurologische Klinik, Universitätsklinikum, Heinrich-Heine-Universität-Düsseldorf, Moorenstraße 5, D-40225, Düsseldorf, Germany

    Philipp Goertz, Mario Siebler & Ulrich Junghans

  3. Anatomisches Institut, Nussallee 10, D-53115, Bonn, Germany

    Stephan L. Baader

Authors
  1. Stephanie Linke
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Corresponding author

Correspondence to Joachim Kappler.

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Stephanie Linke and Philipp Goertz contributed equally.

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Linke, S., Goertz, P., Baader, S.L. et al. Aldolase C/Zebrin II is Released to the Extracellular Space after Stroke and Inhibits the Network Activity of Cortical Neurons. Neurochem Res 31, 1297–1303 (2006). https://doi.org/10.1007/s11064-006-9169-9

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  • DOI: https://doi.org/10.1007/s11064-006-9169-9

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