gms | German Medical Science

27. Wissenschaftlicher Kongress der Deutschen Hochdruckliga

Deutsche Liga zur Bekämpfung des hohen Blutdrucks – Deutsche Hypertonie Gesellschaft e. V.

26. bis 29.11.2003, Bonn

Combination of laser microdissection and genome-wide expression profiling for the analysis of renal damage in hypertension

Kombination von Laser-Microdissektion und genomweit expression profiling für die Analyse der hypertensiven Nierenschädigung

Meeting Abstract (Hypertonie 2003)

  • R. Kreutz - Charite-Universitätsmedizin, Max-Planck- Institut für molekulare Genetik, Abtei, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin (Berlin, D)
  • presenting/speaker K. Janitz - Charite-Universitätsmedizin, Max-Planck- Institut für molekulare Genetik, Abtei, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin (Berlin, D)
  • M. Wehland - Charite-Universitätsmedizin, Max-Planck- Institut für molekulare Genetik, Abtei, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin (Berlin, D)
  • R. Herwig - Charite-Universitätsmedizin, Max-Planck- Institut für molekulare Genetik, Abtei, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin (Berlin, D)
  • M. Janitz - Charite-Universitätsmedizin, Max-Planck- Institut für molekulare Genetik, Abtei, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin (Berlin, D)

Hypertonie 2003. 27. Wissenschaftlicher Kongress der Deutschen Hochdruckliga. Bonn, 26.-29.11.2003. Düsseldorf, Köln: German Medical Science; 2004. Doc03hochP9

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter: http://www.egms.de/de/meetings/hoch2003/03hoch109.shtml

Veröffentlicht: 11. November 2004

© 2004 Kreutz et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

The Munich Wistar Frömter (MWF) rat represents an inbred genetic model that develops an increased urinary albumin excretion and arterial hypertension. In addition, these animals are characterised by a reduced number of nephrons (30-50%) when compared with other rat strains. In order to elucidate molecular mechanisms involved in the glomerular phenotype we performed a genome-wide analysis of the gene expression patterns in glomerular tissue of the MWF and the normotensive Wistar rat with normal renal function. The glomeruli have been isolated either using the differential sieving method or by a laser microdissection and pressure catapulting (LMPC) technique from histological cryosections. Total RNA isolated from both type of tissue preparations has been labeled and hybridised with the rat GeneChip U34A from Affymetrix. For LMPC analysis the RNA has been additionally amplified using two rounds of in vitro transcription. The initial amount of total RNA isolated with the LMPC technique (based on 20 glomeruli) ranged from 1 to 10 ng per sample. Comparison of gene expression profiles in the glomeruli pellets between the two rat strains resulted in identification of 294 genes, which have been up-regulated and 185 being down-regulated at least 2-fold in the MWF animals. We are currently involved in evaluation of particular candidate genes using real-time PCR. Furthermore, we compared a data set obtained from glomerular tissue obtained by sieving and microdissected glomeruli in order to establish the degree of their correlation. Our data indicate that the combination of laser microdissection together with microarray analysis can be successfully applied for establishment of the genes expression patterns from the histologicaly defined tissue structures such as the glomerulus. Therefore, this method will be important for the molecular analysis of hypertensive target organ damage in both animal and human tissues.