Publication

Gene expression patterns in rat dentate granule cells: Comparison between fresh and fixed tissue

Qin, Y., Heine, VM., Karst, H., Lucassen, PJ. & Joels, M., 30-Dec-2003, In : Journal of Neuroscience Methods. 131, 1-2, p. 205-211 7 p.

Research output: Contribution to journalArticleAcademicpeer-review

Copy link to clipboard

Documents

  • Gene expression patterns in rat dentate granule cells: comparison between fresh and fixed tissue

    Final publisher's version, 318 KB, PDF document

    Request copy

DOI

  • Y Qin
  • VM Heine
  • H Karst
  • PJ Lucassen
  • M Joels

RNA from brain tissue (in particular human brain) can often only be extracted from fixed material. As brain tissue is very heterogeneous with regard to cell type, it is important to obtain RNA from small samples of identified cells. The aim of this study was (a) to generate expression profiles from small yet homogenous samples of fixed brain cells in rat and (b) to verify the reliability of these profiles by comparing them with expression profiles obtained from single fresh neurons of the same cell type. Samples (n = 12) of 50 rat dentate granule cells were isolated, using Laser Microdissection and Pressure Catapulting, from paraformaldehyde fixed, paraffin embedded tissue or from frozen, ethanol fixed tissue. In addition, RNA was extracted under visual control from individual dentate granule cells (n = 12) in hippocampal slices, after electrophysiological recording with patch clamp electrodes. Our data show that RNA was successfully extracted from ethanol fixed sections yielding expression profiles highly comparable to those from non-fixed, single granule cells. RNA extraction from paraformaldehyde fixed, paraffin embedded tissue was less reliable. The present approach validates expression profiling from small amounts of fixed neurons as a powerful tool to investigate molecular processes if fresh tissue is not available. (C) 2003 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)205-211
Number of pages7
JournalJournal of Neuroscience Methods
Volume131
Issue number1-2
Publication statusPublished - 30-Dec-2003
Externally publishedYes

    Keywords

  • laser dissection microscope, hippocampal slice, RNA amplification, dentate gyrus, rat, LASER CAPTURE MICRODISSECTION, PARAFFIN-EMBEDDED TISSUES, IN-SITU HYBRIDIZATION, MESSENGER-RNA, HOUSEKEEPING GENE, BRAIN-TISSUE, AMPLIFICATION, FIXATION, NEURONS, SINGLE

ID: 65920963