Comparison of Wild type and Pofut1-deleted skeletal muscle

Pofut1 is an essential gene that glycosylates proteins containing EGF-like repeats, including Notch Receptors (NotchRs).  Work in mice and in Drosophila has shown that O-fucosylation by Pofut1 is required for NotchR ligands to bind to and activate NotchRs.  As such, Pofut1 deletion in skeletal myofibers allows for an analysis of potential functions and molecular changes of Pofut1 in skeletal muscle that derive from its expression in skeletal myofibers. In this study we compared gene expression profiles between quadriceps muscles in mice where Protein O-fucosyltransferase 1 (Pofut1) was deleted specifically in skeletal myofibers via use of a human skeletal alpha actin Cre transgene (Scre) and a loxP flanked Pofut1 gene (SCreFF) and mice which bore the only the Scre transgene but did not have floxed Pofut1 alleles (SCre++). Muscles were dissected from two month-old mice. We have found that SCreFF mice develop reduced muscle size and strength with age.  These changes are analogous to sarcopenic changes that normally occur in aged muscles, where Pofut1 expression is also reduced.  These phenotypes begin at and beyond 3 months and are very significant by 6 months of age.   The goal of this study was to capture molecular changes that precede the reductions in muscle size and strength, some of which were validated by qRT-PCR. A total of six samples in two groups were analyzed for this study. The first group is three replicates of skeletal α actin cre/Pofut1flox/flox quadriceps. The second group is three replicates of Skeletal α actin cre/Pofut1 wild type quadriceps.