. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. DISSOCIATED MUSCLE FIBERS OF APLYSIA 111. buccal muscles were dissected from the animal, teased into thin strips, and incubated for 2-4 h at 28°C in Instant Ocean containing 10 mAI HEPES (pH , adjusted with NaOH), collagenase (Sigma Type I). soybean trypsin inhibitor (Sigma Type I-S), and 1 //g/ml leupeptin Figure 1. Typical dissociated buccal muscle fiber. At the right is a patch electrode pointing to the fiber. Scale: calibration marks are apart. muscles 5-HT produces no contractile response itself but do
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. DISSOCIATED MUSCLE FIBERS OF APLYSIA 111. buccal muscles were dissected from the animal, teased into thin strips, and incubated for 2-4 h at 28°C in Instant Ocean containing 10 mAI HEPES (pH , adjusted with NaOH), collagenase (Sigma Type I). soybean trypsin inhibitor (Sigma Type I-S), and 1 //g/ml leupeptin Figure 1. Typical dissociated buccal muscle fiber. At the right is a patch electrode pointing to the fiber. Scale: calibration marks are apart. muscles 5-HT produces no contractile response itself but does potentiate ACh-elicited contractions (Ram el a/.. 1981). MCGs are active during feeding (Weiss et a/.. 1978). Lesion of serotonergic neurons changes (although does not completely block) feeding motor activity (Rosen et a/.. 1983, 1989). Thus, the modulatory effect of the se- rotonergic MCG neurons on feeding muscles appears to have an important role in feeding arousal. Although previous experiments on mechanisms me- diating the modulatory effect of 5-HT on buccal muscles have suggested roles for cyclic AMP (Mandelbaum, 1980: 1983, 1984a) and calcium (Ram 1984b; Ram and Parti. 1985), the cellular targets of these me- diators have not been determined. For example, it is un- known whether cyclic AMP or calcium modify membrane mechanisms such as ion channels or change the sensitivity or activity of contractile proteins. For studying effects on contractile proteins, this laboratory developed a skinned muscle preparation, which is described elsewhere (Ram and Patel, 1989). To study membrane mechanisms, we developed a dissociated muscle fiber preparation. This paper describes the contractile properties of this disso- ciated muscle preparation, including its modulatory re- sponse to 5-HT, and demonstrates its suitability for patch clamp analysis of single channel and whole cell ionic cur- rents. Preliminary descriptions of some of these data have appeared previousl
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
