Cyclin D1 messenger RNA is induced in microglia rather than neurons following transient forebrain ischaemia.
Following 30 min of forebrain ischaemia in the rat, delayed neuronal death occurs in the CA1 sector of the hippocampus within two to three days, whereas neurons in other selectively vulnerable regions, such as the dorsolateral striatum, die within 6-12 h. In this study, we investigated cyclin D1 expression, which codes for a regulatory protein in cell cycle regulation, but it is also induced in sympathetic neurons undergoing programmed cell death. Cyclin D1 messenger RNA could not be detected by in situ hybridization techniques in brains of control rats, but was found at one and two days after ischaemia in regions of the dorsolateral striatum with neuronal degeneration. DNA fragmentation in this region, identified by the terminal transferase biotinylated-UTP nick end labelling (TUNEL) procedure, was observed from 6 h after ischaemia onward. In the hippocampus, increased levels of cyclin D1 messenger RNA were found at two and three days after ischaemia in the striatum pyramidale of the CA1 sector. This expression was associated with the occurrence of neuronal damage and TUNEL-stained neurons. By seven days cyclin D1 messenger RNA was found in hardly any brain structure. There was no temporospatial overlap of cyclin D1 expression with the expression of the immediate-early genes c-fos, c-jun, and mkp-1, a result which is clearly distinct from findings in sympathetic ganglion neurons undergoing programmed cell death. These results do not suggest a role for cyclin D1 in neuronal cell death following transient forebrain ischaemia. The similarity of the cyclin D1 expression profile with that of the microglia-specific CR3 complement receptor beta-subunit messenger RNA, and the results of combined in situ hybridization and microglia-specific immunohistochemistry suggest that microglia are the source of cyclin D1 messenger RNA in the postischaemic brain. Since cyclin D1 codes for a critical regulatory protein for progression of the G0 to G1 phase in the cell cycle and we did not observe prominent occurrence of DNA fragmentation in microglial cells in the hippocampus at time points when cyclin D1 messenger RNA was found, we suggest that cyclin D1 induction is involved in the onset of microglial cell proliferation.
Wiessner
C
C
Max-Planck-Institute for Neurological Research, Department for Experimental Neurology, Cologne, Germany.
Brink
I
I
Lorenz
P
P
Neumann-Haefelin
T
T
Vogel
P
P
Yamashita
K
K
D000818:Animals; D000918:Antibody Specificity; D001157:Arterial Occlusive Diseases / Q000503:physiopathology; D001253:Astrocytes / Q000502:physiology; D001483:Base Sequence; D001710:Biotin; D016923:Cell Death / Q000502:physiology; D019938:Cyclin D1; D016213:Cyclins / Q000235:genetics*; D004249:DNA Damage; D003856:Deoxyuracil Nucleotides; D005786:Gene Expression Regulation / Q000502:physiology; D017781:Genes, Immediate-Early / Q000502:physiology; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D017403:In Situ Hybridization; D002546:Ischemic Attack, Transient / Q000235:genetics* / Q000503:physiopathology*; D008297:Male; D017628:Microglia / Q000276:immunology / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D015513:Oncogene Proteins / Q000235:genetics*; D016548:Prosencephalon / Q000098:blood supply / Q000166:cytology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011951:Receptors, Complement / Q000235:genetics; D013194:Staining and Labeling
D016213:Cyclins; D003856:Deoxyuracil Nucleotides; D018840:HSP70 Heat-Shock Proteins; D015513:Oncogene Proteins; D012333:RNA, Messenger; D011951:Receptors, Complement; D019938:Cyclin D1; D001710:Biotin