Deepali Mathur
University of Valencia, Spain
Title: Evaluation of housekeeping genes in rat neurons and oligodendrocyte progenitor cells treated with cerebrospinal fluid of multiple sclerosis subjects
Biography
Biography: Deepali Mathur
Abstract
Gene expression studies employing real-time PCR has become an intrinsic part of biomedical research. Appropriate normalization of target gene transcript(s) based on stably expressed housekeeping genes is crucial in individual experimental conditions to obtain accurate results. In multiple sclerosis (MS), several gene expression studies have been undertaken, however, the suitability of housekeeping genes to express stably in this disease is not yet explored. The present study aims to evaluate the expression stability of a panel of housekeeping genes in rat neurons and oligodendrocyte progenitor cells (OPCs) treated with cerebrospinal fluid derived from MS patients. For neurons, reference genes chosen from literature were quantified by real time PCR and their expression stability was assessed using geNorm and NormFinder algorithms. For OPCs, reference genes were chosen from microarray data and their stability was assessed using similar algorithms. Both algorithms revealed transferrin receptor (Tfrc) and microglobulin beta-2 (B2m) the most stable genes followed by ribosomal protein L19 (Rpl19) in neurons whereas beta-actin (ActB) and glyceraldehyde-3-phosphate-dehydrogenase (Gapdh) the most fluctuated ones. On the contrary, geNorm revealed mitochondrial ribosomal protein L19 (Mrpl19), hypoxanthine phosphoribosyl transferase (Hprt) and microglobulin ï¢2 (B2m) as the best housekeeping genes and Gapdh the least stable in CSF exposed OPCs. Altogether our data demonstrate the significance of pre-validation of housekeeping genes for accurate normalization and indicates Tfrc and B2m as best endogenous controls in neurons and Mrpl19 and Hprt in OPCs treated with CSF from MS patients. ActB and Gapdh are not recommended in gene expression studies related to current one.