[The synchronization of the rhythm of protein synthesis in hepatocyte cultures occurs during conditioning of the medium with ganglioside GM1 accumulation in the cells: an immunohistochemical study].
We studied the kinetics of proteins synthesis in the cultures of hepatocytes on collagen-coated slides in medium 199 enriched with 0.2 mg/ml albumin and 0.5 microgram/ml insulin and devoid of bovine serum. Circahoral fluctuations of proteins synthesis intensity were found in the monolayer cultures and sparse cultures in a conditioned medium. No protein synthesis rhythm was expressed in a fresh medium after repeated washing of the cultures. Addition of micromolar concentrations of gangliosides GM1 or GD1a to the medium revealed the rhythm in washed cultures. Addition of GT1b or GM3 to the medium did not synchronize the oscillations: the kinetics of protein synthesis did not differ from that in the control space cultures in a fresh medium without exogenous gangliosides. Accumulation of gangliosides GM1 and GM3 in the hepatocytes in vitro upon conditioning of a serum-free culture medium was shown using the indirect immunocytochemical method. The effect was found in dense monolayer cultures and in cultures with separated cells in a conditioned medium. The protein synthesis rhythm was found in such cultures. Gangliosides are weakly expressed in most cells of a repeatedly washed 24-hour monolayer and washed sparse cultures. No protein synthesis rhythm was found in washed cultures. Similar changes in the dynamics of the culture medium conditioning, accumulation of gangliosides in cells and rhythmic activity of cells population confirm the concept of the synchronizing role of gangliosides.
Brodskiĭ
V Ia
VIa
Nechaeva
N V
NV
Zvezdina
N D
ND
Novikova
T E
TE
Gvazava
I G
IG
Fateeva
V I
VI
D000818:Animals; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D005677:G(M1) Ganglioside / Q000378:metabolism*; D005679:G(M3) Ganglioside / Q000378:metabolism; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D010507:Periodicity; D014176:Protein Biosynthesis; D051381:Rats; D013997:Time Factors
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D005679:G(M3) Ganglioside; D005677:G(M1) Ganglioside