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通过原位杂交技术对正常、非整倍体及脑损伤小鼠大脑中淀粉样前体蛋白mRNA进行神经解剖定位与定量分析。

Neuroanatomical localization and quantification of amyloid precursor protein mRNA by in situ hybridization in the brains of normal, aneuploid, and lesioned mice.

作者信息

Bendotti C, Forloni G L, Morgan R A, O'Hara B F, Oster-Granite M L, Reeves R H, Gearhart J D, Coyle J T

机构信息

Johns Hopkins University School of Medicine, Baltimore, MD 21205.

出版信息

Proc Natl Acad Sci U S A. 1988 May;85(10):3628-32. doi: 10.1073/pnas.85.10.3628.

Abstract

Amyloid precursor protein mRNA was localized in frozen sections from normal and experimentally lesioned adult mouse brain and from normal and aneuploid fetal mouse brain by in situ hybridization with a 35S-labeled mouse cDNA probe. The highest levels of hybridization in adult brain were associated with neurons, primarily in telencephalic structures. The dense labeling associated with hippocampal pyramidal cells was reduced significantly when the cells were eliminated by injection of the neurotoxin ibotenic acid but was not affected when electrolytic lesions were placed in the medial septum. Since the gene encoding amyloid precursor protein has been localized to mouse chromosome 16, we also examined the expression of this gene in the brains of mouse embryos with trisomy 16 and trisomy 19 at 15 days of gestation. RNA gel blot analysis and in situ hybridization showed a marked increase in amyloid precursor protein mRNA in the trisomy 16 mouse head and brain when compared with euploid littermates or with trisomy 19 mice.

摘要

通过与35S标记的小鼠cDNA探针进行原位杂交,淀粉样前体蛋白mRNA定位在正常和实验性损伤的成年小鼠脑以及正常和非整倍体胎儿小鼠脑的冰冻切片中。成年脑中最高水平的杂交与神经元相关,主要位于端脑结构中。当通过注射神经毒素鹅膏蕈氨酸消除海马锥体细胞时,与海马锥体细胞相关的密集标记显著减少,但当在内侧隔放置电解损伤时,该标记不受影响。由于编码淀粉样前体蛋白的基因已定位到小鼠16号染色体,我们还检测了妊娠15天时16三体和19三体小鼠胚胎脑中该基因的表达。RNA凝胶印迹分析和原位杂交显示,与整倍体同窝小鼠或19三体小鼠相比,16三体小鼠头部和脑中淀粉样前体蛋白mRNA显著增加。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e354/280267/657c03b7c233/pnas00262-0368-a.jpg

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