Myonecrosis induced by guanidine in the mouse isolated phrenic nerve diaphragm preparation

1. The myonecrosis induced by guanidine in the mouse phrenic nerve diaphragm preparation was investigated using both light microscopy and myographic recordings. Preparations were incubated with 10 mM guanidine for 60 min in the absence and presence of electrical stimulation. At the end of this period, the drug was washed out and the nutritive medium replaced with fixative solution to prevent morphological artefacts. 2. Guanidine produced a triphasic change in the amplitude of twitch tension evoked indirectly through the motor nerve. This response consisted of an initial facilitation followed by a neuromuscular blockade and a secondary facilitatory effect after removal of the drug. 3. Morphological analysis of the muscle showed various structural alterations of the fibers, including the presence of very dark swollen cells with or without small clear vacuoles, delta lesions with densely or loosely clumped myofibrils, irregular clear spaces, indistinct masses of degraded myofibrils, and, in extreme cases, "ghost" cells. All of these effects were attributed to the presence of high cytosolic calcium concentrations. 4. Pretreatment with tetrodotoxin (TTX, 3.13 microM) diminished but did not prevent the guanidine-induced morphological abnormalities in the muscle cells. This finding suggests that TTX can interfere to a certain extent with the influx of guanidine into muscle fibers through sodium channels. 5. An attempt was made to correlate the myographic findings with the muscle morphological alterations seen after guanidine removal.