The expression of MrrSOD, i-NOS and e~NOS mRNAs in KO mice gingiva

KO小鼠牙龈MrrSOD、i-NOS和e~NOS mRNA的表达

• 批准号: 12671783
• 负责人: WATANABE Hiroki
• 金额: $ 2.5万
• 依托单位: TOKYO DENTAL COLLEGE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671783/
• 关键词: FREE RADICAL; NOS; SOD; nRNA; KNOCKOUT MICE; GINGIVA; in situ HYBRIDIZATION; IMMUNOHIBIOCHEMISTRY; in situ hybridiazation

Free radicals and superoxide are known to be involved in metabolism and transport. And the expression of enzymes related to these has been evaluatedin some cells. To delineate the kinetics of these enzymes in gingiva of normal, SODITg(Transgerliic), NOS2KO(Knockout) and NOS3KO mice we evaluated Mn-SOD, i-NOS and eHNOS mRNAs expression in gingiva after perfusion fixation and paraffin embedding. We used in situ hybridization with specific probe to evaluate mRNA expression and Western blotting. We also examined immunohtstochemistry with nitrotyrosine antibody for protein expression. Mn-SOD expression was weakly positive in normal gingiva but strongly positive in NDS2 KO and NOS3 KO gingiva. Expression of i-NOS was absent in normal gingiva but positive in SOD1 Tg and NOS3KO gingiva. Expression of e-NOS was weakly positive in rtormal gingivabut strongly positive in NOS 2 KO gingiva.The results of the experiment on knockout mice suggested that superoxide dismutase increased in the absence of i-NOS production, and i-NOS increased in a compensatory manner in the absence of e-NOS production in gingiva.

已知自由基和超氧化物参与新陈代谢和运输。与此相关的酶的表达已经在一些细胞中得到了评估。为了探讨这些酶在正常、变性、NOS2KO和NOS3KO小鼠牙龈中的动态变化，我们检测了灌流固定和石蜡包埋后，这些酶在牙周组织中的表达。用特异性探针原位杂交法和Western blotting检测mRNA的表达。我们还用硝基酪氨酸抗体进行了免疫组织化学检测蛋白表达。正常牙周组织中有弱阳性表达，而NDS2KO和NOS3KO牙周组织中有强阳性表达。I-NOS在正常牙周组织中无表达，而在SOD1TG和NOS3KO牙龈组织中呈阳性表达。E-NOS在正常牙周呈弱阳性表达，而在NOS2-KO牙周呈强阳性表达。基因敲除小鼠的实验结果表明，在没有i-NOS产生的情况下，超氧化物歧化酶升高，而在牙龈中没有e-NOS产生的情况下，i-NOS呈代偿性增加。

项目成果

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SAbe.N.笠原 MAmang M.Yoshii。

Hiroki Watanabe et al.: "Colony-stimu latin g factor-1(CSF-1) res cues ost eobl ast attachm ent, surv ival and sor ting of beta-a ctin mRNA in the toothle ss(t1-oste ope trotic) muta tion in the rat"Int.J.Develop.Biol.. 44. 201-207 (2000)

Hiroki Watanabe 等人：“集落刺激因子 1 (CSF-1) 修复了牙齿中 ost eoblast 的附着、存活和 β-肌动蛋白 mRNA 的分选 (t1-oste ope trotic)

S.Mitarashi et al.: "Temporomandibular joint ankylosis"J.Craniomandibular practice (Cranio). 20 No.1. 67-71 (2002)

S.Mitarashi 等人：“颞下颌关节强直”J.Craniomandibular 实践 (Cranio)。

Hiroki Watanabe et al.: "Colony-stimulating factor-1 (CSF-1) rescues osteoblast attachment, survival and sorting of beta-actin mRNA in the toothless (t1-osteopetrotic)"Int.J.Develop.Biol.. 44. 201-207 (2000)

Hiroki Watanabe 等人：“集落刺激因子 1 (CSF-1) 可挽救无牙 (t1-osteopetrotic) 中成骨细胞的附着、存活和 β-肌动蛋白 mRNA 的分选”Int.J.Develop.Biol.. 44。

Hiroki Watanabe et al.: "Colony-stimulating factor-1(CFS-1) rescues osteoblast attachment, survival and sorting of beta-actin mRNA in the toothless(tl-osteopetrotic) mutation in the rat"Int.J.Develop.Biol.. 44. 201-207 (2000)

Hiroki Watanabe 等人：“集落刺激因子 1 (CFS-1) 可挽救大鼠无牙（tl-骨质疏松）突变中成骨细胞的附着、存活和 β-肌动蛋白 mRNA 的分选”Int.J.Develop.Biol