Proteomic Identification of Immunoproteasome Accumulation in Formalin-Fixed Rodent Spinal Cords with Experimental Autoimmune Encephalomyelitis
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- 作者：Mohit Raja Jain ; Qing Li ; Tong Liu ; Joseph Rinaggio ; Amit Ketkar ; Vincent Tournier ; Kiran Madura ; Stella Elkabes ; Hong Li
- 刊名：Journal of Proteome Research
- 出版年：2012
- 出版时间：March 2, 2012
- 年：2012
- 卷：11
- 期：3
- 页码：1791-1803
- 全文大小：590K
- 年卷期：v.11,no.3(March 2, 2012)
- ISSN：1535-3907
文摘
Clinically relevant formalin-fixed and paraffin-embedded (FFPE) tissues have not been widely used in neuroproteomic studies because many proteins are presumed to be degraded during tissue preservation. Recent improvements in proteomics technologies, from the 2D gel analysis of intact proteins to the 鈥渟hotgun鈥?quantification of peptides and the use of isobaric tags for absolute and relative quantification (iTRAQ) method, have made the analysis of FFPE tissues possible. In recent years, iTRAQ has been one of the main methods of choice for high throughput quantitative proteomics analysis, which enables simultaneous comparison of up to eight samples in one experiment. Our objective was to assess the relative merits of iTRAQ analysis of fresh frozen versus FFPE nervous tissues by comparing experimental autoimmune encephalomyelitis (EAE)-induced proteomic changes in FFPE rat spinal cords and frozen tissues. EAE-induced proteomic changes in FFPE tissues were positively correlated with those found in the frozen tissues, albeit with 50% less proteome coverage. Subsequent validation of the enrichment of immunoproteasome (IP) activator 1 in EAE spinal cords led us to evaluate other proteasome and IP-specific proteins. We discovered that many IP-specific (as opposed to constitutive) proteasomal proteins were enriched in EAE rat spinal cords, and EAE-induced IP accumulation also occurred in the spinal cords of an independent mouse EAE model in a disability score-dependent manner. Therefore, we conclude that it is feasible to generate useful information from iTRAQ-based neuroproteomics analysis of archived FFPE tissues for studying neurological disease tissues.
<h4>Keywords: h4> immunoproteasome; experimental autoimmune encephalomyelitis; iTRAQ; formalin-fixed paraffin-embedded tissue; biomarker; mass spectrometry
<h4>Keywords: h4> immunoproteasome; experimental autoimmune encephalomyelitis; iTRAQ; formalin-fixed paraffin-embedded tissue; biomarker; mass spectrometry