1993 Fiscal Year Final Research Report Summary
Effect of thyroid hormone on gene expression in the central nervous system of newborn rats
| Project/Area Number |
04671458
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | DOKKYO UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
MATSUZAKI Sigeru DOKKYO UNIV.SCH.MED., PROFESSOR, 医学部, 教授 (60008604)
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| Co-Investigator(Kenkyū-buntansha) |
KOIBUCHI Noriyuki DOKKYO UNIV.SCH.MED., ASSISTANT, 医学部, 助手 (80234681)
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| Project Period (FY) |
1992 – 1993
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| Keywords | THYROID HORMONE / RAT CEREBELLUM / GENE EXPRESSION / CYTOCHROME C OXIDASE / c-erbA alpha2 / DNA SEQUENCE / NORTHERN BLOTTING |
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| Research Abstract |
In 1992 we started to construct cDNA library from the cerebellum of newborn rats. Methimazole dissolved in drinking water was given to pregnant rats from 15th day of their pregnancy and continued giving till the 16th day after delivery. On 16th day after birth newborn rats were killed and their cerebella were dissected out and mRNA in the organ was extracted. cDNA library was constructed from mRNA extracted from the cerebellum of control animals using lambda gt10 vector. The differential plaque hybridization was performed using ^<32>P-labelled antisense DNA which had been prepared from the mRNA extracted from the control group and hypothyroid group. We found several clones of which expression may possibly be affected by thyroid states. These clones were subcloned in plasmid to be produced in a large amount.
In 1993 the clones mentioned above were analyzed for their nucleotide sequences. Using the cDNA subcloned as a probe, nothern blot analysis was performed for the mRNA from the control and hypothyroid group. The genes whose expression was suppressed a mitochondrial gene in the central nervous system is influenced by thyroid states.
We also examined the effects of thyroid hormone in rat brain on the expression of other genes which had already been identified. The quantitative in situ hybridization showed that c-erbA alpha 2mRNA in the hypothalamic paraventricular nucleus was increased after thyroidectomy of adult male rats. The increase was prevented by thyroxine. These findings show that the expression of some genes in rat brain is augmented and that of others is suppressed.
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