Preparation of tetramethylrhodaminyl-phalloidin and uptake of the toxin into short-term cultured hepatocytes by endocytosis.
A fluorescent phallotoxin with high photostability, tetramethylrhodaminyl-phalloidin (Rh-phalloidin), has been prepared. The affinity of this compound to rabbit muscle actin has been determined to be about 6 times lower than that of phalloidin. In freshly isolated hepatocytes the internalized fluorescent toxin stains the cellular actin. In contrary, there is no actin staining visible in cultured hepatocytes. Short-term cultured hepatocytes (5 h of culturing) incorporate the toxin by endocytosis; it is kept sealed in the endocytotic vesicles, which are usually found accumulated at the sites where cells touch after reaggregation.
D000199:Actins / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D004705:Endocytosis; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D009842:Oligopeptides / Q000378:metabolism*; D010590:Phalloidine / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012235:Rhodamines
D000199:Actins; D009842:Oligopeptides; D012235:Rhodamines; D010590:Phalloidine; C005358:tetramethylrhodamine