Techniques for the Measurement of Lipoprotein Lipase Messenger RNA

This chapter describes the measurement of lipoprotein lipase (LPL) mRNA levels in adipose tissue and muscle. Many investigators measure LPL in human biopsy samples, where only a small amount of tissue is available, requiring sensitive measurement techniques. In addition, the description of low levels of LPL expression in tissues such as liver, where it may play an important role in remnant lipoprotein uptake (1 ), also requires sensitive methods. Therefore, this chapter will concentrate on the use of competitive reverse transcriptase-polymerase chain reaction (RT-PCR) for the quantitation of relatively low levels of LPL mRNA. Only brief mention will be made of Northern blotting and RNase protection assays, since these established methods are well described in commonly available laboratory manuals.