REGULATION OF GLYCOSPHINGOLIPID BIOSYNTHESIS IN MOUSE TISSUES

University Of Georgia, Athens GA

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Abstract

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Transcript analysis data from our previous studies with Dr. Campo were in agreement with previous studies conducted by her laboratory. However, previously published expression data obtained for ST6Gal1 using end-point PCR indicated an increase in expression with the age of harvested pituitaries, while data collected using our qRT-PCR platform indicated a decrease or no change in expression under similar conditions. We investigated differences between the two technologies and designed primers for qRT-PCR, which were in similar locations as the primers used by the Campo group. The conclusion of these studies supported data we acquired using our original primer sets and suggests that the lack of sensitivity inherent in the end-point PCR technique is the likely reason for the lack of agreement between data sets. Several future experiments are planned using a restricted gene set and the potential for switching to a tissue culture cell line instead of using whole tissue samples.