CLONING OF MECHANOSENSITIVE RECEPTORS

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CELL IS ALIVE UNDER CHEMICAL,LIGHT AND MECHANICAL STIMULI AND EXTRACELLULAR STRESS.BIOLOGICAL PERCEPTION OF MECHANICAL STRESS INCLUDES HEARING,BALANCE,TOUCH SENSATION,DETECTION OF MOVEMENT OF MUSCLE AND TENDON,AND NOTICE OF VOLUME EXPANSION.MECHANICAL STIMULI INCLUDE FRICTION,PRESSURE AND STRETCH.CELLS ADAPT TO THESE STIMULI BY INTRACELLULAR METABOLISM.CHANGE IN ACTIVITY OF ION CHANNELS RESPONDING TO MECHANICAL STRESS IS CLEAR.HOWEVER,IT IS NOT CLEAR THAT ION CHANNELS ARE MECHANOSENSITIVE RECEPTORS THEMSELVES OR NOT.ONE REASON WHY THEY ARE NOT FOUND IS THAT THEY ARE TINY AND SCARCE IN CELLS.IN SPITE OF THIS SITUATION RECENT DEVELOPMENT OF RESEARCH TECHNIQUES IN MOLECULAR BIOLOGY MAKES IT POSSIBLE TO SOLVE THE PROBLEM.OUR STUDY APPLIED SEVERAL STRATE GIES AS FOLLOWS : 1) CLONING OF MECHANOSENSITIVE RECEPTOR cDNAs BY POLYMERASE CHAIN REACTION BASED METHODS.HOWEVER,WE IDENTIFIED cDNAs ALREADY REPORTED ; 2) DEVELOPMENT OF APPARATUS OF POSITIVE PRESSURE ON CULTURE CELLS.THIS APPARATUS IS UNDER USE TO ONGOING STUDY ; 3) IN-GEL PROTEIN KINASE ASSAY METHOD AND 4) mRNA DIFFERENTIAL DISPLAY METHOD ARE UNDER USE TO STUDY CHANGE IN INTRACELLULAR SECOND MESSENGER SYSTEMS AND TO FIND NOVEL GENES WHICH RESPOND TO EXTRACELLULAR POSITIVE PRESSURE.