1998 Fiscal Year Final Research Report Summary
Molecular Histochemical Analysis on Nuclear Distribution of Transcription Regulatory Factors and the Transcriptional Activity of the Genes.
| Project/Area Number |
09670018
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Nagasaki University |
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Principal Investigator |
IZUMI Shin-ichi Nagasaki University School of Medicine, Assistant, 医学部, 助手 (40264246)
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| Co-Investigator(Kenkyū-buntansha) |
SHIN Masashi Nagasaki University School of Medicine, Assistant, 医学部, 助手 (80145226)
KOJI Takehiko Nagasaki University School of medicine, Professor, 医学部, 教授 (30170179)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Transcription / Nucleus / Prolactin / Growth Hormone / in situ hybridization / Southwestern / Placenta / Pituitary |
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| Research Abstract |
1. The optimun conditions of a combination treatment with exogenous or endogenous DNase and followed with proteinase K (0.02 unit/ml) for leucocytes were successful in DNA in situ hybridization histochemistry using labeled-hunan X chromoosome DNA (Xp21, Xq13, Xq21) probes. This revealed that active and inactive X chromiatin DNA in cells were localized in nuclear euchromatin and heterochromtin, respectively.
2. In rat anterior pituitary GH3 culture cells and rat anterior pituitary tissue sections Southwestern histochemistry (SWH), which localizes DNA binding proteins, was performed. The method demonstrated that Pit-1 transcription regulatory factor participating in activation of growth hormone (GH) gene was localized in nuclei of GH cells, whereas PREB transcription regulatory factor in suppression was not localized in the nuclei, and that Pit-1 also in activation of prolactin (PRL) gene as well as PREB were local ized in nuclei of PRL cal Is.
3. Rats were intraperitoneally injected 10 Nm GH releasing hormone or 10 nM scmatostatin or control saline 4 hours before pituitary gland dissection. The anterior pituitary tissue sections from them were stained for Pit-1 and PREB by SWH No marked difference of intranuclear distribution and staining intensity could be recognized among them. When Pit-1 is neutralized in GH3 cells, whether transcription of GH gene is suppressed has not been confirmed yet.
4. In rat placentas at around 14 day of gestation, some similar trophoblasts started localizing nuclear Pit-1 and cytcplasmic PRL.As gestational day goes on, number and staining intensity for both positive trophoblasts increased. However, at around 19 day of gestation, Pit-1 was also seen in the cytoplasm and staining intensity for PRL decreased in the trophoblasts by immuno-histochemistry. It is suggested that distributing nuclear Pit-1 in the cells of rat placentas may be associated in functional differentiation of trophoblasts that produce PRL-like substances.
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