2007 Fiscal Year Final Research Report Summary
Quantification of transcripts in testes preparation and its application to other tissue sections
| Project/Area Number |
18591801
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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 |
Obstetrics and gynecology
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| Research Institution | Okayama University |
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Principal Investigator |
SASAKI Junzo Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor (30093686)
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| Co-Investigator(Kenkyū-buntansha) |
MIKI Yukari Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Researcher (70397876)
YAMADA Teruo Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Professor (00033225)
KOSAKA Jun Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Associate Professor (40243216)
FUJITA Hirofumi Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Professor (20423288)
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| Project Period (FY) |
2006 – 2007
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| Keywords | Anatomy / Cell and tissue / Testis / In situ hybridization / Gene / mRNA / PERF 15 / Quantification of signals |
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| Research Abstract |
Testis was a suitable organ for quantifying the ISH signals because germ cells undergo synchronized development and show stage-specific gene expression. Previously, we used ribosomal RNA as the hybridizable RNA in paraffin sections and could easily analyze ISH signals expressed with digoxygenin-labeled probes quantitatively, through "posterization" of the images. In the present project, we applied this method to analyze the quantification of transcripts, PERF 15 mRNA. PERF 15 was a 15 kDa protein found in the perforatorium of the sperm head and identified as a.testis lipid binding protein. To determine an absolute quantity of transcripts in tissue sections, we further analyzed the signals by a Confocal Laser Scanning microscope with the use of a tyramide signal amplification system.
The peak of PERF 15 mRNA expression was found in diplotene spermatocytes, and the amount of PERF 15 mRNA w as greatest in late pachytene and diplotene primary spermatocytes and early spermatids, followed by early pachytene primary spermatocytes, and then late spermatids. PERF 15 may be involved in the events leading to meiotic division, in which apoptosis is also involved.
The present study could help our attempt to determine the concentration of mRNA in the tissue section. In this protocol, we made glass slides on which a tissue section and row of oligonucleotides spots were placed to determine the concentration of mRNA in a tissue section. These spots, 110 pm in diameter, consisted of 1 nl of sense and antisense mRNA sequences, from 0-1 × 10^<-6> M to 500 M, and we attempted to compare the signal intensity in the tissue section with the signal intensity in the spots containing known concentrations of the given mRNA.
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[Journal Article] Alpha-lipoic acid suppresses 6-hydroxydopamine-induced ROS generation and apoptosis through the stimulation of glutathione synthesis but not by the expression of heme oxygenase-12008
Author(s)
Hirofumi, Fujita, Masahiko, Shiosaka, Tetsuya, Ogino, Yuya, Okimura, Toshihiko, Utsumi, Eisuke, F., Sato, Reiko, Akagi, Masayasu, Inoue, Kozo, Utsumi, Junzo, Sasaki
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Journal Title
Brain researc 1206
Pages: 1-12
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Histological observation of the development of follicles and follicular atresia in immature ovaries2007
Author(s)
Takagi, K., Yamada, T., Miki, Y., Umegaki, T., Nishimura, M., Sasaki, J
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Journal Title
Acta Med Okayama 61(5)
Pages: 283-298
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Gene profiles during root canal treatment in experimental rat periapical lesions2007
Author(s)
Zulema, Rosalia, Arias, Martinez, Koji, Naruishi, Keisuke, Yamashiro, Fumio, Myokai, Teruo, Yamada, Kaori, Matsuura, Naoko, Namba, Hideo, Arai, Junzo, Sasaki, Yoshimitsu, Abiko, Shogo, Takashiba
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Journal Title
Journal of Endodontics 33
Pages: 936-943
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Quantification of PERF 15 mRNA in tissue sections from rat testes2006
Author(s)
Takashi, Kogami, Yukari, Mild, Teruo, Yamada, Teruo, Umegaki, Makoto, Nishimura, Takashi, Amo, Jun, Kosaka, Junzo, Sasaki
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Journal Title
Acta Histochemica et Cytocchemica 39(6)
Pages: 183-192
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Flow cytometric analysis of Ca^<2+>induced membrane permeability transition of isolated rat liver mitochondria2003
Author(s)
Teruo, Umegaki, Yuya, Okimura, Hirofumi, Fujita, Hiromi, Yano, Jitsuo, Akiyama, Masayasu, Inoue, Kozo, Utsumi, Junzo, Sasaki
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Journal Title
Journal of Clinical Biochemistry and Nutrition 42
Pages: 35-44
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Regulation of 5-aminolevulinic acid-dependent protoporphyrin IX accumulations in human histiocytic lymphoma U937 cells2003
Author(s)
Yuya, Okimura, Hirofumi, Fujita, Tetsuya, Ogino, Keiji, Inoue, Hiromi, Yano, Tatsuji, Yasuda, Masayasu, Inoue, Kozo, Utsumi, Junzo, Sasaki
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Journal Title
Physiol. Chemistry & Phys. and Med. NMR (In press)
Pages: 39
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Role of α-lipoic acid on mitochondrial functions, redox metabolism and apoptosis of cells2007
Author(s)
Hirofumi, Fujita, Youya, Okimura, Toshihiko, Utsumi, Eisuke, Sato, Kozo, Utsumi, Junzou, Sasaki
Organizer
International Conference on Food Factors for health promotion
Place of Presentation
Kyoto
Year and Date
20071127-1201
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Regulation of of 5-aminolevulinic acid-mediated protoporphyrin IX-accumulation and its photodynamic action in U937 cells2007
Author(s)
Hirofumi, Fujita, Yuya, Okimura, Tetsuya, Ogino, Tatsuji, Yasuda, Koichi, Abe, Kozo, Utsumi, Junzo, Sasaki
Organizer
International Conference on Food Factors for, health promotion
Place of Presentation
Kyoto
Year and Date
20071127-1201
Description
「研究成果報告書概要(欧文)」より
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[Presentation] DNA microarray analysis on genes expressed in periapical lesion2007
Author(s)
Arias Martinez, ZR., Yamashiro, K., Naruishi, K., Matsuura, K., Myokai, F., Yamada, T., Sasaki, J., Arai, H., Abiko, Y., Takashiba, S
Organizer
International Association for Dental Research & American Association for Dental Research
Place of Presentation
New Orleans, Louisiana
Year and Date
20070321-24
Description
「研究成果報告書概要(欧文)」より
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