Dense connective tissue repair : the intricacies of monitoring actin isoforms in native tissue models in vitro
Gwillim, Eran C.
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In preparation to quantify beta-actin and alpha-smooth muscle actin content in loaded and unloaded native rat tail tendon organ culture models, several methodologies were developed in the present paper. Since alpha-smooth muscle actin has not been monitored in tendon in vitro, developing the methods to culture and quantitatively detect alpha-smooth muscle actin, the protein marker for fibroblast to myofibroblast transition were developed. In addition, quantifying the amount of alpha-smooth muscle actin mRNA, using reverse transcription polymerase chain reaction (RT-PCR) assays was investigated and novel oligonucleotide primers were developed. A method to identify four highly conserved actin isoforms; beta cytoplasmic, gamma cytoplasmic, alpha- smooth muscle and gamma smooth muscle actin, in a tissue environment was created.