Scanning Microscopy


The rabbit masseter is composed of mainly fast fatiguable type IIB fibers which give a positive mATPase reaction at pH 9.3. To study the effect of fatigue on the distribution of some crucial elements, the masseter was stimulated electrically for 45 minutes. The contralateral muscle served as a control. Control and experimental tissues were rapidly excised, and frozen in isopentane cooled to -150°C in liquid nitrogen. Sections were cut in a cryostat and prepared by freeze-drying. Water content of fatigued muscle was significantly higher than controls: 79.3% versus 77.3%. The fatigued state was verified by glycogen depletion viewed in PAS-stained sections. Ice crystal artifacts were more prominent in stimulated muscle, suggesting cytoplasmic edema and/or a change in water state. Element analysis of myofibers, using an scanning electron microscope equipped with a computer assisted energy dispersive X-ray spectrometer was performed at 15 kV for 100 seconds. Counts were recorded for Na, Mg, P, S, Cl, K and Ca ( Mg and Ca counts were too low to evaluate). Counts were converted to concentrations using a gelatin element standard. The most striking finding in fatigued muscle was a decrease in K concentration (33%) and an increase in Na (100%) and Cl (75%). P and S values were slightly lower. An interpretation of these findings is made in relation to a) changes in water structure and b) conformational changes in myofibril macromolecules.

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