Neurons from fetal rat brains contain functional luteinizing hormone chorionic gonadotropin receptors
Neurons from fetal rat brains contain functional luteinizing hormone chorionic gonadotropin receptors
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DOI：
10.1095/biolreprod56.5.1071
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发表时间：
1997-05-01
影响因子：
3.6
通讯作者：
Rao, CV
中科院分区：
文献类型：
--
作者：
AlHader, AA;Lei, ZM;Rao, CV
成年和新生大鼠的大脑含有功能性LH/hCG受体。这些发现使我们假设胎鼠大脑也可能含有这些受体。为了验证这一假设，我们从19天大的胎鼠脑中分离神经元，并将其培养在化学成分确定的无血清培养基中。逆转录聚合酶链反应扩增出预期的256碱基对大小的LH/hCG受体片段，该片段可与Southern印迹中的全长LH/hCG受体cDNA杂交。北方印迹显示，神经元含有一个主要的2.6 kb和一个次要的4.3 kb的转录本。免疫细胞化学染色显示神经元含有LH/hCG受体。Western免疫印迹显示，神经元含有一个80 kDa的受体蛋白，在培养的第3天增加到最高水平，然后逐渐下降，直到培养的第9天。在高度纯化的hCG的存在下培养神经元3天，导致神经突突起和总细胞蛋白的生长呈剂量依赖性增加，并且与相应对照组中的值相比，DNA片段减少。在最大有效hCG浓度下，神经突承载细胞的数量增加了53%，总细胞蛋白增加了60%，DNA片段减少了31%。总之，这是第一个研究证明LH/hCC受体的存在和hCG的神经营养作用在胎鼠脑神经元。这些结果表明，局部产生的促性腺激素可能在胎儿脑的生长和发育中发挥作用。
Adult and neonatal rat brains contain functional LH/hCG receptors. These findings have led us to hypothesize that the fetal rat brain may also contain these receptors. To test this hypothesis, we isolated neurons from 19-day-old fetal rat brains and cultured them in chemically defined serum-free medium. The reverse transcription polymerase chain reaction amplified an expected 256-base pair size LH/hCG receptor fragment that could hybridize with a full-length LH/hCG receptor cDNA in Southern blotting. Northern blotting demonstrated that neurons contained a major 2.6 kilobase (kb) and a minor 4.3 kb transcript. Immunocytochemistry demonstrated that the neurons contained LH/hCG receptor immunostaining. Western immunoblotting showed that neurons contained an 80-kDa receptor protein that increased to a maximal level on Day 3 of culture and then gradually decreased until the 9th day of culture. Culturing neurons for 3 days in the presence of highly purified hCG resulted in a dose-dependent increase in the outgrowth of neurite processes and total cellular protein and a decrease in DNA fragmentation as compared to values in the corresponding controls. At the maximally effective hCG concentration, the number of neurite-bearing cells was increased by 53% and the total cellular protein by 60%, and DNA fragmentation decreased by 31%. In summary, this is the first study to demonstrate the presence of LH/hCC receptors and neurotrophic effects of hCG in fetal rat brain neurons. These findings imply that locally produced gonadotropins may possibly play a role in the growth and development of the fetal brain.