Cyclin dependent kinase inhibitors prevent apoptosis of postmitotic mouse motoneurons.
Recent evidence suggests that apoptosis in post-mitotic neurons involves an aborted attempt of cells to re-enter the cell cycle which is characterized by increased expression of cyclins, such as cyclin D1, prior to death. However, such cyclins activation prior to apoptotic cell death remains controversial. Many neurological disorders are characterized by neuronal loss, particularly amyotrophic lateral sclerosis (ALS). ALS is a motoneuronal degenerative condition in which motoneuron loss could be due to an inappropriate return of these cells in the cell cycle. In the present study, we observed that deprivation of neurotrophic factor in purified motoneuron cultures induces an apoptotic pathway. After neurotrophic factor withdrawal, DAPI (4,6-diamidin-2-phenylindol dichlorohydrate) staining revealed the presence of nuclear condensation, DNA fragmentation, and perinuclear apoptotic body. Similarly, release of apoptotic microparticles and activation of caspases-3 and -9 were observed within the first hours following neurotrophic factor withdrawal. Next, we tested whether inhibition of cell cycle-related cyclin-dependent kinases (cdks) can prevent motoneuronal cell death. We showed that three cdk inhibitors, olomoucine, roscovitine and flavopiridol, suppress the death of motoneurons. Finally, we observed early increases in cyclin D1 and cyclin E expression after withdrawal of neurotrophic factors. These findings support the hypothesis that after removal of trophic support, post-mitotic neuronal cells die due to an attempt to re-enter the cell cycle in an uncoordinated and inappropriate manner.
Appert-Collin
A
A
Université Louis Pasteur, Faculté de Pharmacie, UMR7175-LC1, BP 24, F-67401 Illkirch Cedex, France.
Hugel
B
B
Levy
R
R
Niederhoffer
N
N
Coupin
G
G
Lombard
Y
Y
André
P
P
Poindron
P
P
Gies
J P
JP
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000378:metabolism; D019927:Cyclin E / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D007701:Kinetin / Q000494:pharmacology; D051379:Mice; D008938:Mitosis; D009046:Motor Neurons / Q000166:cytology* / Q000187:drug effects*; D009414:Nerve Growth Factors / Q000172:deficiency; D010880:Piperidines / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011687:Purines / Q000494:pharmacology; D000077546:Roscovitine
D019927:Cyclin E; D005419:Flavonoids; D009414:Nerve Growth Factors; D010880:Piperidines; D047428:Protein Kinase Inhibitors; D011687:Purines; D000077546:Roscovitine; D019938:Cyclin D1; C077990:alvocidib; C090046:olomoucine; D018844:Cyclin-Dependent Kinases; D007701:Kinetin