Abstract
A new improvement in the production of cell-free models of Physarum for studying the physiology of cytoplasmic actomyosin contraction in vivo is described. Cytoplasmic actomyosin fibrils were enriched in thin-spread plasmodia of Physarum by allowing them to spread on agar containing 2% ethanol. Polarizing microscopy of plasmodia before and after extraction showed a high density of enriched birefringent actomyosin fibrils. Plasmodia not treated with alcohol but prepared similarly showed lower density of birefringent actomyosin fibrils before and even less density after extraction. It seemed that alcohol treatment enriched and stabilized the actomyosin fibrils and that they were less affected by the process of extraction. It was shown, that the fibrils contain actin and their distribution was studied by fluorescent microscopy after staining with rhodamine-labelled phalloidin. These results may help to correlate the birefringent picture of actomyosin fibrils with their fluorescence and thus improve the cell-free model for reactivation studies.
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