Terminin (Tp 63/60), a novel cell senescence-related protein, is present in the aging human hippocampus.
We have previously identified a cell senescence-related, detergent-insoluble cytoplasmic protein referred to as terminin, and have presented the development-dependent modification of terminin polypeptides in rat brain. In the present study, we characterize biochemically and immunocytochemically terminin expression in human hippocampus. By immunoblotting with a monoclonal antibody (mab 1.2) against terminin, we identified a 63 kDa polypeptide (Tp63) in the detergent-insoluble fraction of human brain proteins, which can be identified by either one- or two-dimensional electrophoresis. The 63 kDa protein in immunoprecipitates of human hippocampus was found to share a peptide pattern with the 60 kDa terminin (Tp60) previously found in rat brain, as demonstrated by in situ peptide mapping, suggesting that the human 63 kDa protein is a homologue of the rat Tp60. By immunocytochemistry, terminin immunoreactivity was demonstrated throughout the neuronal layers in the human hippocampus. The pyramidal cell layer in hippocampus proper and subiculum, as well as the granule cells in the dentate gyrus, gave rise to immunoreactivity for terminin. It was observed that in the cornu Ammonis, the highest terminin immunoreactivity was present in the pyramidal neurons of CA1. At the cellular level terminin was localized intraneuronally, as demonstrated by confocal microscopy of double staining with both mab 1.2 and an anti-neurofilament antibody. Taken together, the present study establishes that terminin is expressed in abundance in aging human CNS neurons.
Yang
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G
Bloomfield Centre for Research in Aging, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, Department of Medicine, McGill University, Montréal, Qué., Canada.
Wang
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E
D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007525:Isoelectric Focusing; D009474:Neurons / Q000378:metabolism; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D010449:Peptide Mapping; D011233:Precipitin Tests; D051381:Rats; D016707:Tissue Fixation
D000911:Antibodies, Monoclonal; D009687:Nuclear Proteins; C083046:terminin