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AND MMP-2 EXPRESSION IN CAPILLARIES ISOLATED FROM OVERLOADED SKELETAL MUSCLES: LASER-CAPTURE MICRODISSECTION STUDY
1 Kinesiology and Health Sciences, York University, Toronto, ON, Canada
* To whom correspondence should be addressed. E-mail: thaas{at}yorku.ca.
Under physiological non-hypoxic conditions angiogenesis can be driven by mechanical forces. However, due to the limitations of the specific gene expression analysis of microvessels from in vivo experiments, the mechanisms regulating the coordinated expression of angiogenic factors implicated in the process remain intangible. In this study, the technique of laser capture microdissection (LCM) was adapted for the study of angiogenesis in skeletal muscles. Using a combination of LCM and real-time quantitative PCR it was demonstrated that capillary endothelial cells produce matrix metalloproteinase-2 (MMP-2) and that mechanical stretch of capillaries within muscle tissue markedly increased MMP-2 mRNA (2.5 fold increase vs. control; P<0.05). In addition, we showed that transcription factor HIF-1
expression was 13.5- fold higher in capillaries subjected to stretch compared to controls (P<0.05). These findings demonstrate the feasibility of this approach to study angiogenic gene regulation and provide novel evidence of HIF-1 induction in stretched capillary endothelial cells.
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