title: Mechano-regulation of gap junction communications between tendon cells is dependent on the magnitude of tensile strain.
abstract: Large magnitudes of mechanical strain applied to tendon cells induce catabolic and inflammatory responses, whereas a moderate level of strain promotes anabolism. Gap junction intercellular communication (GJIC) plays an essential role in these responses, however direct regulation of GJIC by mechanical loading has not been characterised in detail. Here, we show that the GJIC between tenocytes are enhanced or inhibited depending on the magnitude of the tensile strain. The GJIC was analysed using fluorescence loss in photobleaching (FLIP), combined with a molecular diffusion model. Intercellular and intracellular transport of fluorescence tracer molecules, calcein, across multiple cells through the gap junctions was evaluated by determining the intercellular and intracellular diffusion coefficients of calcein. It was demonstrated that the intercellular diffusion coefficient was significantly higher when the cells were subjected to a physiological static tensile strain (4%) for 1 h, but significantly lower when subjected to a strain with non-physiological amplitude (8%). The intracellular diffusion coefficient was not altered by the application of static strain at any level. Connexin 43 proteins were localised within cytoplasm and at cell-cell boundaries in no strained state and were also localised near cell nuclei by the 4% strain, but the localisation was reduced by the 8% strain. The findings suggest that the increase in GJIC in response to 4% strain involves opening of gap junction pores via mechanotransduction events of tenocytes, whereas the inhibition in response to 8% strain involves mechanical disruption of the junctions.
authors.affiliation: Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan. Electronic address: e.maeda@eng.hokudai.ac.jp.
authors.forename: Eijiro
authors.initials: E
authors.lastname: Maeda
authors.affiliation: Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan.
authors.forename: Toshiro
authors.initials: T
authors.lastname: Ohashi
mesh_terms: D000818:Animals; D001692:Biological Transport; D002450:Cell Communication; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D004058:Diffusion; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005452:Fluoresceins; D036681:Fluorescence Recovery After Photobleaching; D005456:Fluorescent Dyes; D017629:Gap Junctions / Q000378:metabolism* / Q000648:ultrastructure; D015870:Gene Expression; D008297:Male; D040542:Mechanotransduction, Cellular; D011817:Rabbits; D013710:Tendons / Q000166:cytology / Q000378:metabolism*; D013718:Tensile Strength / Q000502:physiology
chemical_list: D018031:Connexin 43; D005452:Fluoresceins; D005456:Fluorescent Dyes; C007740:fluorexon
keywords: Diffusion coefficient; FLIP; Gap junction intercellular communication; Tendon; Tenocytes