An Optimal Protocol to Analyze the Rat Spinal Cord Proteome

  1. Martin-Rojas, T. 1
  2. Barroso, G. 2
  3. Gil-Dones, F. 1
  4. Alonso-Orgaz, S. 1
  5. Scott-Taylor, J. 3
  6. Barderas, M.G. 1
  7. Avila, G. 3
  8. Moral-Darde, V. 2
  9. Vivanco, F. 45
  1. 1 Department of Vascular Pathophysiology, Hospital Nacional de Paraplejicos (HNP), SESCAM, Toledo.
  2. 2 Proteomics Unit, Hospital Nacional de Paraplejicos (HNP), SESCAM, Toledo.
  3. 3 Sensorimotor Function Group, Hospital Nacional de Paraplejicos (HNP), SESCAM, Toledo.
  4. 4 Department of Immunology, Fundacion Jimenez Diaz, Madrid.
  5. 5 Department of Biochemistry and Molecular Biology I, Universidad Complutense, Madrid.
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Revista:
Biomarker Insights

ISSN: 1177-2719,  1177-2719

Año de publicación: 2009

Volumen: 4

Páginas: BMI.S2965

Tipo: Artículo

DOI: 10.4137/BMI.S2965 GOOGLE SCHOLAR lock_openAcceso abierto editor

Otras publicaciones en: Biomarker Insights

Resumen

Since the function of the spinal cord depends on the proteins found there, better defing the normal Spinal Cord Proteome is an important and challenging task. Although brain and cerebrospinal fluid samples from patients with different central nervous system (CNS) disorders have been studied, a thorough examination of specific spinal cord proteins and the changes induced by injury or associated to conditions such as neurodegeneration, spasticity and neuropathies has yet to be performed. In the present study, we aimed to describe total protein content in the spinal cord of healthy rats, employing different proteomics tools. Accordingly, we have developed a fast, easy, and reproducible sequential protocol for protein extraction from rat spinal cords. We employed conventional two dimensional electrophoresis (2DE) in different pH ranges (eg. 4–7, 3–11 NL) combined with identification by mass spectrometry (MALDI-TOF/TOF), as well as first dimension protein separation combined with Liquid Chromatography Mass Spectrometry/Mass Spectrometry (LC-MS/MS) to maximise the benefits of this technology. The value of these techniques is demonstrated here by the identification of several proteins known to be associated with neuroglial structures, neurotransmission, cell survival and nerve growth in the central nervous system. Furthermore this study identified many spinal proteins that have not previously been described in the literature and which may play an important role as either sensitive biomarkers of dysfunction or of recovery after Spinal Cord Injury.