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Synchronization of Plodia interpunctella lepidopteran cells and effects of 20‐hydroxyecdysone
Author(s) -
Siaussat D.,
Mottier V.,
Bozzolan F.,
Porcheron P.,
Debernard S.
Publication year - 2004
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.0962-1075.2004.00475.x
Subject(s) - 20 hydroxyecdysone , biology , cell cycle , flow cytometry , microbiology and biotechnology , cell division , lepidoptera genitalia , plodia interpunctella , ecdysone receptor , mitosis , complementary dna , botany , cell , biochemistry , pyralidae , nuclear receptor , gene , transcription factor
We have investigated the molecular and cellular mechanisms involved in the control of insect cell cycle by 20‐hydroxyecdysone (20E) using the IAL‐PID2 cell line established from imaginal wing discs of Plodia interpunctella . We first defined conditions for use of hydroxyurea, a reversible inhibitor of DNA synthesis, in order to synchronize the IAL‐PID2 cells in their division cycle. A high degree of synchrony was reached when cells were exposed to two consecutive hydroxyurea treatments at 1 m m for 36 h spaced 16 h apart. Under these conditions, flow cytometry analysis demonstrated that 20E at 10 −6 m induced an inhibition of cell growth by an arrest of 90% of the cells in G2/M phase. Using cDNA probes specifically designed from E75 and HR3 nuclear receptors of Plodia interpunctella , we showed that PiE75 and PHR3 were highly induced by 20E through S and G2 phases with maximal enhancement just before the G2/M arrest of cells. These findings suggest that PiE75 and PHR3 could be involved in a 20E‐induced genetic cascade leading to G2/M arrest.