|Budget Amount *help
¥1,800,000 (Direct Cost : ¥1,800,000)
Fiscal Year 1992 : ¥300,000 (Direct Cost : ¥300,000)
Fiscal Year 1991 : ¥1,500,000 (Direct Cost : ¥1,500,000)
In this study, monoclonal antibodies to the heavy-chain isoforms of myosins in human brain and liver were produced for the development of identification methods of human tissues. Three monoclonal anti-human cerebrum myosin antibodies (HBM 1,4 and 6) and two monoclonal anti-human liver myosin antibodies (HLM 1 and 4) each having distinctive tissue-specificity were obtained by immunizing mice with the myosin purified from the human cerebrum and liver, respectively. And immunoblotting of the heavy-chain isoforms of myosins in human nonmuscle tissues (the cerebrum, cerebellum, liver, spleen, kidney and platelet), and the aorta and cardiac muscle as controls were carried out using these antibodies.
Consequently, 2-4 heavy-chain isoforms of myosins were found in these tissues. Myosin heavy-chain of the cerebrum contained four isoforms which showed the molecular masses of about 200,198,196 and 194 kDa, and that of the liver had 198,196 and 194 kDa isoforms. Since 200 and 198 kDa isoforms were mainly found in the brain, they were thought to be brain-type, while 196 and 194 kDa were commonly found in the nonmuscle tissues, therefore, they were seemed to be nonmuscle-type. On the other hand, myosin heavy-chain isoforms in the liver mainly consisted of common nonmuscle-type. Monoclonal antibody HBM 1 recognized specifically the brain-type isoforms of myosin heavy-chain. Therefore, HBM 1 seemed to be useful to discriminate the brain tissue from others by the immunological procedures. Indeed, brain-type isoforms of myosin heavy chain-could be detected in blood sera from bodies with cerebral contusion by sandwich ELISA using HBM 1.