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Establishment of a protocol for the gene expression analysis of laser microdissected rat kidney samples with affymetrix genechips

Establishment of a protocol for the gene expression analysis of laser microdissected rat kidney samples with affymetrix genechips

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STEMMER, Kerstin, Heidrun ELLINGER-ZIEGELBAUER, Kerstin LOTZ, Hans-Jürgen AHR, Daniel R. DIETRICH, 2006. Establishment of a protocol for the gene expression analysis of laser microdissected rat kidney samples with affymetrix genechips. In: Toxicology and applied pharmacology. 217(1), pp. 134-142. ISSN 0041-008X. Available under: doi: 10.1016/j.taap.2006.07.012

@article{Stemmer2006Estab-8072, title={Establishment of a protocol for the gene expression analysis of laser microdissected rat kidney samples with affymetrix genechips}, year={2006}, doi={10.1016/j.taap.2006.07.012}, number={1}, volume={217}, issn={0041-008X}, journal={Toxicology and applied pharmacology}, pages={134--142}, author={Stemmer, Kerstin and Ellinger-Ziegelbauer, Heidrun and Lotz, Kerstin and Ahr, Hans-Jürgen and Dietrich, Daniel R.} }

First publ. in: Toxicology and applied pharmacology 217 (2006), 1, pp. 134-142 application/pdf eng Establishment of a protocol for the gene expression analysis of laser microdissected rat kidney samples with affymetrix genechips 2011-03-24T17:39:44Z 2006 Stemmer, Kerstin deposit-license 2011-03-24T17:39:44Z Stemmer, Kerstin Laser microdissection in conjunction with microarray technology allows selective isolation and analysis of specific cell populations, e.g., preneoplastic renal lesions. To date, only limited information is available on sample preparation and preservation techniques that result in both optimal histomorphological preservation of sections and high-quality RNA for microarray analysis. Furthermore, amplification of minute amounts of RNA from microdissected renal samples allowing analysis with genechips has only scantily been addressed to date. The objective of this study was therefore to establish a reliable and reproducible protocol for laser microdissection in conjunction with microarray technology using kidney tissue from Eker rats p.o. treated for 7 days and 6 months with 10 and 1 mg Aristolochic acid/kg bw, respectively. Kidney tissues were preserved in RNAlater or snap frozen. Cryosections were cut and stained with either H&E or cresyl violet for subsequent morphological and RNA quality assessment and laser microdissection. RNA quality was comparable in snap frozen and RNAlater-preserved samples, however, the histomorphological preservation of renal sections was much better following cryopreservation. Moreover, the different staining techniques in combination with sample processing time at room temperature can have an influence on RNA quality. Different RNA amplification protocols were shown to have an impact on gene expression profiles as demonstrated with Affymetrix Rat Genome 230_2.0 arrays. Considering all the parameters analyzed in this study, a protocol for RNA isolation from laser microdissected samples with subsequent Affymetrix chip hybridization was established that was also successfully applied to preneoplastic lesions laser microdissected from Aristolochic acid-treated rats. Ellinger-Ziegelbauer, Heidrun Ellinger-Ziegelbauer, Heidrun Ahr, Hans-Jürgen Dietrich, Daniel R. Lotz, Kerstin Ahr, Hans-Jürgen Lotz, Kerstin Dietrich, Daniel R.

Dateiabrufe seit 01.10.2014 (Informationen über die Zugriffsstatistik)

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