PMID- 7615657
OWN - NLM
STAT- MEDLINE
DCOM- 19950822
LR  - 20231213
IS  - 0021-9533 (Print)
IS  - 0021-9533 (Linking)
VI  - 108 ( Pt 4)
DP  - 1995 Apr
TI  - Reappearance and long-term maintenance of connexin32 in proliferated adult rat 
      hepatocytes: use of serum-free L-15 medium supplemented with EGF and DMSO.
PG  - 1347-57
AB  - Intercellular communication, especially gap junctional communication, is thought 
      to be one of the highly differentiated functions of hepatocytes. In primary 
      cultures of rat hepatocytes, it has been considered that the maintenance and the 
      reinduction of differentiated functions is very difficult. In the present study, 
      we succeeded in inducing the gap junctional protein connexin32 (Cx32) in adult 
      rat hepatocytes cultured in serum-free L-15 medium supplemented with epidermal 
      growth factor (EGF) and dimethylsulfoxide (DMSO). When the hepatocytes were 
      cultured in L-15 medium supplemented with 20 mM NaHCO3 and 10 ng/ml EGF in a 5% 
      CO2:95% air incubator, the cells proliferated. Fluorescence immunocytochemistry 
      showed spots immunoreactive to Cx32 on the cell membranes between adjacent cells 
      until day 3, but only a few Cx32-positive spots were found after day 4. Western 
      and northern blot analyses also showed that the amounts of both the protein and 
      mRNA of Cx32 in the cells decreased with time in culture. However, when the cells 
      were treated with 2% DMSO from day 4, the immunoreactive spots reappeared on the 
      cell membranes from day 6 and both their number and intensity gradually 
      increased. The reappearance of Cx32 was accompanied by increases in both the 
      protein and mRNA of Cx32. Furthermore, the expression of Cx32 was well 
      maintained, together with extensive gap junctional intercellular communication, 
      for more than 4 weeks. In addition, ultrastructurally, many gap junctional 
      structures were observed between the hepatocytes, and the antibodies to Cx32 were 
      shown to bind to those structures. This culture system may be useful for studies 
      of the reconstruction of the gap junctional structure, the intracellular pathways 
      of the proteins, and the regulation of synthesis and processing in differentiated 
      hepatocytes.
FAU - Kojima, T
AU  - Kojima T
AD  - Department of Pathology, Sapporo Medical University School of Medicine, Japan.
FAU - Mitaka, T
AU  - Mitaka T
FAU - Paul, D L
AU  - Paul DL
FAU - Mori, M
AU  - Mori M
FAU - Mochizuki, Y
AU  - Mochizuki Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Cell Sci
JT  - Journal of cell science
JID - 0052457
RN  - 0 (Connexins)
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Isoquinolines)
RN  - 62229-50-9 (Epidermal Growth Factor)
RN  - 9654F8OVKE (lucifer yellow)
RN  - YOW8V9698H (Dimethyl Sulfoxide)
SB  - IM
MH  - Animals
MH  - Blotting, Northern
MH  - Blotting, Western
MH  - Cell Communication
MH  - Cells, Cultured
MH  - Connexins/analysis/*biosynthesis
MH  - Culture Media, Serum-Free
MH  - Dimethyl Sulfoxide/*pharmacology
MH  - Epidermal Growth Factor/*pharmacology
MH  - Fluorescent Dyes
MH  - Gap Junctions/*physiology/ultrastructure
MH  - Gene Expression
MH  - Isoquinolines
MH  - Liver/*cytology/drug effects/*physiology
MH  - Male
MH  - Microscopy, Fluorescence
MH  - Microscopy, Immunoelectron
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Gap Junction beta-1 Protein
EDAT- 1995/04/01 00:00
MHDA- 1995/04/01 00:01
CRDT- 1995/04/01 00:00
PHST- 1995/04/01 00:00 [pubmed]
PHST- 1995/04/01 00:01 [medline]
PHST- 1995/04/01 00:00 [entrez]
AID - 10.1242/jcs.108.4.1347 [doi]
PST - ppublish
SO  - J Cell Sci. 1995 Apr;108 ( Pt 4):1347-57. doi: 10.1242/jcs.108.4.1347.