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Choroid plexus (ChP), a vascularized secretory epithelium located in all brain ventricles, plays critical roles in development, homeostasis and brain repair. Reverse transcription quantitative real-time PCR (RT-qPCR) is a popular and useful technique for measuring gene expression changes and also widely used in ChP studies. However, the reliability of RT-qPCR data is strongly dependent on the choice of reference genes, which are supposed to be stable across all samples. In this study, we validated the expression of 12 well established housekeeping genes in ChP in 2 independent experimental paradigms by using popular stability testing algorithms: BestKeeper, DeltaCq, geNorm and NormFinder.  Rer1  and  Rpl13a  were identified as the most stable genes throughout mouse ChP development, while  Hprt1  and  Rpl27  were the most stable genes across conditions in a mouse sensory deprivation experiment. In addition,  Rpl13a ,  Rpl27  and  Tbp  were mutually among the top five most stable genes in both experiments. Normalisation of  Ttr  and  Otx2  expression levels using different housekeeping gene combinations demonstrated the profound effect of reference gene choice on target gene expression. Our study emphasized the importance of validating and selecting stable housekeeping genes under specific experimental conditions.

Assessment of common housekeeping genes as reference for gene expression studies using RT-qPCR in mouse choroid plexus