Open AccessInhibition of peroxisomal degradation by 3‐methyladenine (3MA) in primary cultures of rat hepatocytes
Author(s) -
Kondo Kazumasa,
Makita Takashi
Publication year - 1997
Publication title -
the anatomical record
Language(s) - English
Resource type - Journals
eISSN - 1097-0185
pISSN - 0003-276X
DOI - 10.1002/(sici)1097-0185(199704)247:4<449::aid-ar2>3.0.co;2-m
Subject(s) - peroxisome , collagenase , catalase , biology , microbiology and biotechnology , cell culture , andrology , chemistry , biochemistry , enzyme , receptor , medicine , genetics
Background A significant reduction in peroxisomes has been demonstrated in primary cultures of rat hepatocytes. This report demonstrates that 3‐methyladenine (3MA), a potent inhibitor of autophagy, inhibits this effect. Methods Hepatocytes from male Wistar rats were isolated by a two‐step in situ perfusion technique using collagenase and were cultured in Williams E medium. After a 2‐hr attachment period (day 0 of culture), the cells were treated with 200 μM bezafibrate (BF), a peroxisome proliferator, and 5 mM 3MA for 3 days. The cells in the culture dish were fixed in situ, stained for catalase, and embedded in Poly/Bed 812. The number and size of peroxisomes in electron micrographs were analyzed morphometrically. Results After 3 days of culture, the number of peroxisomes had decreased to 30% of the day 0 level. However, the day 0 level was maintained by treatment with 3MA. In BF‐treated cells, many autophagosomes were observed, and peroxisomes had proliferated significantly, although they did not exceed the day 0 level. In cells treated with a combination of 3MA and BF, the number and size of peroxisomes had increased remarkably. Conclusions These results suggest that 3MA is effective in maintaining both the number and size of peroxisomes in the course of primary cultures of rat hepatocytes. Suppression of peroxisome proliferation by treatment with BF may be regulated by autophagic/lysosomal degradation. Anat. Rec. 247:449–454, 1997. © 1997 Wiley‐Liss, Inc.