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Patch area (a) is inversely correlated with pipette resistance (R) according to the relation a = 12.6(1/R + 0.018) [B. Sakmann and E. Neher, in Single Channel Recordings, B. Sakmann and E. Neher, Eds. (Plenum, New York, 1983), pp. 37-51]. We carefully monitored patches for visible membrane in the pipette tip and for unusual suction durations. All patches for which extensive membrane (other than the small omega-shaped dome) was visible in the pipette tip or that required inordinately long suction for a seal were excluded from the study. There were relatively few of these, with a similar incidence in each group, and nearly all were excluded, in any case, by the criterion of a minimum seal resistance of 15 gigohm. In general, the suction time to seal was at least as rapid in the aged group as in the others, as indicated by the relatively high yield in this group (12).
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To determine whether altered sensitivity to Bay K 8644 could account for aspects of these changes, we also measured the mean channel open time (14) during the step to -30 mV in patches used for the I- V analysis (Fig. 2B) and at -70 mV during the repolarization period (ROs) after steps to +10 mV in all patches in which ROs were present. In these multichannel patches, open time was most accurately measured at -30 mV and at -70 mV, when there were few simultaneous openings. The mean ± SEM values for open time at -30 mV were: young adult, 6.5 ± 1.4 ms; mid-aged, 6.5 ± 1.5 ms; and aged, 5.7 ± 0.8 ms; and at -70 mV (ROs) were: young adult, 5.5 ± 0.6 ms; mid-aged, 5.0 ± 0.5 ms; and aged, 5.7 ± 0.7 ms. These values do not differ statistically, indicating that Bay K 8644 affected the different age groups similarly. This conclusion is further supported by the lack of age difference in voltage dependence (Fig. 2B), because a shift in voltage dependence is also a hallmark of DHP agonists (14).
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We thank R. Gray and D. Johnston for their assistance with the zipper slice preparation, D. Kaiser for his efforts in conducting the behavioral studies, N. M. Porter for critical review and input, and L. Lowery for assistance in preparing the manuscript. Supported in part by grants AG04542 and AG10836 from the National Institute on Aging and by a grant from Bayer, Inc.
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