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原代神经元和星形胶质细胞培养物中雄激素结合蛋白基因表达的研究。

Demonstration of androgen-binding protein gene expression in primary neuronal and astrocyte cultures.

机构信息

The Department of Pediatrics, and The Curriculum in Neurobiology, University of North Carolina, Chapel Hill, North Carolina; and Clinical Research Institute of Montreal, Montreal, Quebec, Canada.

出版信息

Mol Cell Neurosci. 1993 Oct;4(5):432-9. doi: 10.1006/mcne.1993.1054.

Abstract

Testicular androgen-binding protein (ABP) and liver (plasma) sex hormone-binding globulin (SHBG) are extracellular carrier proteins. They are encoded by the same gene and have the identical primary amino acid sequence. The ABP/SHBG gene is also expressed in rat brain; immunoreactive neurons are located throughout the male and female brain and immunoreactive fibers are present in the ventral hypothalamus. ABP/SHBG RNA transcripts are synthesized from two promoters, P(1) and P(A). P(1) regulates synthesis of the mRNA that encodes secreted ABP, whereas, a P(A) mRNA encodes a nonsecreted ABP-like protein without steroid binding properties. P(A) and P(1) transcripts are expressed in the brain. In this study, we demonstrated by Northern hybridization analysis that primary neuronal and astrocyte cultures express ABP mRNA. PCR analysis of ABP cDNA revealed that astroglia express transcripts originating from promoters P(1) and P(A), whereas, neuronal cDNAs contain primarily PA transcripts. These ABP cDNA-hybridizing products represent mRNAs that encode secreted ABP and the nonsecreted ABP-like protein. Furthermore, it was shown by immunohistochemistry that cultured astrocytes contain immunoreactive ABP. Western blots of astrocyte proteins yielded immunoreactive ABP migrating as 38,000-, 34,000-, and 24,060-M(r) proteins. The sizes of these proteins do not correspond to the protein-encoding properties of P(1) or P(A) ABP mRNA or any known alternatively processed ABP mRNAs, suggesting that they are derived by proteolytic processing of ABP or the ABP-like protein (46,000 Da). The function of the ABP gene products in brain is not obvious. We speculate that ABP may act as a carrier protein for an unknown ligand in axonal transport.

摘要

睾丸雄激素结合蛋白(ABP)和肝脏(血浆)性激素结合球蛋白(SHBG)是细胞外载体蛋白。它们由同一个基因编码,具有相同的一级氨基酸序列。ABP/SHBG 基因也在大鼠脑中表达;免疫反应性神经元存在于雄性和雌性大脑的各个部位,免疫反应性纤维存在于腹侧下丘脑。ABP/SHBG RNA 转录本由两个启动子 P(1)和 P(A)合成。P(1)调节编码分泌型 ABP 的 mRNA 的合成,而 P(A)mRNA 编码一种无类固醇结合特性的非分泌型 ABP 样蛋白。P(A)和 P(1)转录本在脑中表达。在这项研究中,我们通过 Northern 杂交分析证实原代神经元和星形胶质细胞培养物表达 ABP mRNA。ABP cDNA 的 PCR 分析显示,星形胶质细胞表达源自启动子 P(1)和 P(A)的转录本,而神经元 cDNA 主要包含 PA 转录本。这些 ABP cDNA 杂交产物代表编码分泌型 ABP 和非分泌型 ABP 样蛋白的 mRNA。此外,免疫组织化学显示培养的星形胶质细胞含有免疫反应性 ABP。星形胶质细胞蛋白的 Western blot 显示出迁移率为 38,000、34,000 和 24,060-M(r)的免疫反应性 ABP。这些蛋白质的大小与 P(1)或 P(A)ABP mRNA 或任何已知的替代加工 ABP mRNA 的蛋白编码特性不对应,表明它们是由 ABP 或 ABP 样蛋白(46,000 Da)的蛋白水解加工产生的。ABP 基因产物在脑中的功能尚不清楚。我们推测 ABP 可能作为轴突运输中未知配体的载体蛋白发挥作用。

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