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大鼠骨骼肌出生后发育过程中胶质细胞源性神经营养因子(GDNF)和神经营养因子表达的定量分析。

Quantitative analyses of expression of GDNF and neurotrophins during postnatal development in rat skeletal muscles.

作者信息

Nagano Masatoshi, Suzuki Hidenori

机构信息

Department of Pharmacology, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.

出版信息

Neurosci Res. 2003 Apr;45(4):391-9. doi: 10.1016/s0168-0102(03)00010-5.

Abstract

Neurotrophic factors are thought to be critically involved in formation and maintenance of the neuromuscular system. To know precise expression levels of these factors in the muscles during the postnatal period, we developed competitive RT-PCR and two-site enzyme immunoassay and quantitatively measured neurotrophic factors in the rat gastrocnemius and soleus muscles during the postnatal development. mRNAs of glial cell line-derived neurotrophic factor (GDNF) in the gastrocnemius and the soleus muscles were expressed in the highest amount among the neurotrophic factors at birth and dramatically decreased in the first 3 months, while GDNF proteins substantially existed at 3 months of age. Neurotrophin-3 and brain-derived neurotrophic factor in the gastrocnemius muscle kept constant expression in mRNA and protein during the postnatal period. In contrast, mRNA of neurotrophin-4 increased in the first 2 weeks. In the soleus muscles all the neurotrophic factor proteins increased with age for the first month, contrasting with their expressions in the gastrocnemius. The present results showed that GDNF is constitutively supplied to the neuromuscular junction (NMJ) during postnatal development and into adulthood, suggesting its importance in maintenance of the NMJ. Expression of other neurotrophins was also regulated independently during development possibly according to their own roles in the neuromuscular circuit.

摘要

神经营养因子被认为在神经肌肉系统的形成和维持中起着关键作用。为了了解这些因子在出生后时期肌肉中的精确表达水平,我们开发了竞争性逆转录聚合酶链反应(RT-PCR)和双位点酶免疫测定法,并定量检测了出生后发育过程中大鼠腓肠肌和比目鱼肌中的神经营养因子。腓肠肌和比目鱼肌中胶质细胞源性神经营养因子(GDNF)的mRNA在出生时在神经营养因子中表达量最高,在出生后的前3个月急剧下降,而GDNF蛋白在3月龄时大量存在。腓肠肌中的神经营养因子-3(Neurotrophin-3)和脑源性神经营养因子在出生后时期的mRNA和蛋白表达保持恒定。相比之下,神经营养因子-4(Neurotrophin-4)的mRNA在出生后的前2周增加。在比目鱼肌中,所有神经营养因子蛋白在出生后的第一个月都随年龄增加,这与它们在腓肠肌中的表达情况形成对比。目前的结果表明,GDNF在出生后发育直至成年期都持续供应至神经肌肉接头(NMJ),表明其在维持NMJ中的重要性。其他神经营养因子的表达在发育过程中也可能根据它们在神经肌肉回路中的自身作用而独立调节。

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