Cyclin D1 and Cdk4 protein induction in motor neurons after transient spinal cord ischemia in rabbits.
OBJECTIVE
The mechanism of spinal cord injury has been thought to be related to the vulnerability of spinal motor neuron cells against ischemia. However, the mechanisms of such vulnerability are not fully understood. We hypothesized that spinal motor neurons might be lost by programmed cell death and investigated a possible mechanism of neuronal death by detection of double-strand breaks in genomic DNA and immunohistochemical analysis for cyclin D1 and cyclin-dependent kinases (Cdk) 4.


METHODS
We used a rabbit spinal cord ischemia model with a balloon catheter. Spinal cord was removed at 8 hours and 1, 2, and 7 days after 15 minutes of transient ischemia, and histological changes were studied with hematoxylin-eosin staining. In situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL), DNA fragment with gel electrophoresis, Western blot analysis for cyclin D1 and Cdk4, and temporal profiles of cyclin D1 and Cdk4 immunoreactivity were investigated.


RESULTS
Most motor neurons were preserved until 2 days but were selectively lost at 7 days of reperfusion. Immunocytochemistry showed positive TUNEL selectively at 2 days of reperfusion in spinal motor neuron nuclei. Typical ladders of oligonucleosomal DNA fragments were detected at 2 days of reperfusion. Immunoreactivity of cyclin D1 and Cdk4 proteins was induced selectively at 8 hours in motor neuron nuclei, which eventually died.


CONCLUSIONS
These results indicate that induction of cyclin D1 and Cdk4 may be implicated in programmed cell death change after transient spinal cord ischemia in rabbits.
Sakurai
M
M
Department of Thoracic Surgery, Tohoku University School of Medicine, Sendai, Japan.
Hayashi
T
T
Abe
K
K
Itoyama
Y
Y
Tabayashi
K
K
Rosenblum
W I
WI
D000818:Animals; D017209:Apoptosis; D019938:Cyclin D1 / Q000096:biosynthesis*; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases / Q000096:biosynthesis*; D007150:Immunohistochemistry; D009046:Motor Neurons / Q000378:metabolism* / Q000473:pathology*; D011518:Proto-Oncogene Proteins; D011817:Rabbits; D013116:Spinal Cord / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D020760:Spinal Cord Ischemia / Q000378:metabolism* / Q000473:pathology*
D011518:Proto-Oncogene Proteins; D019938:Cyclin D1; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases