Zhang Weiwei, Li Hao, Xing Ziying, Yuan Hongtu, Kindy Mark S, Li Zhenzhong
Department of Anatomy, Shandong University School of Medicine, Jinan, China.
Folia Histochem Cytobiol. 2012 Jul 5;50(2):312-8. doi: 10.5603/fhc.2012.0041.
Both neurotrophins (NTs) and target skeletal muscle (SKM) cells are essential for the maintenance of the function of neurons and nerve-muscle communication. However, much less is known about the association of target SKM cells with distinct NTs on the expression of mRNAs for preprotachykinin (PPT), calcitonin-gene related peptide (CGRP), neurofilament 200 (NF-200), and microtubule associated protein 2 (MAP-2) in dorsal root ganglion (DRG) sensory neurons. In the present study, a neuromuscular coculture model of dissociated dorsal root ganglion (DRG) neurons and SKM cells was established. The morphology of DRG neurons and SKM cells in coculture was observed with an inverted phase contrast microscope. The effects of nerve growth factor (NGF) or neurotrophin-3 (NT-3) on the expression of mRNAs for PPT, CGRP, NF-200, and MAP-2 was analyzed by real time-PCR assay. The morphology of DRG neuronal cell bodies and SKM cells in neuromuscular coculture at different conditions was similar. The neurons presented evidence of dense neurite outgrowth in the presence of distinct NTs in neuromuscular cocultures. NGF and NT-3 increased mRNA levels of PPT, CGRP, and NF-200, but not MAP-2, in neuromuscular cocultures. These results offer new clues towards a better understanding of the association of target SKM cells with distinct NTs on the expression of mRNAs for PPT, CGRP, NF-200 and MAP-2, and implicate the association of target SKM cells and NTs with DRG sensory neuronal phenotypes.
神经营养因子(NTs)和靶标骨骼肌(SKM)细胞对于维持神经元功能和神经 - 肌肉通讯均至关重要。然而,关于靶标SKM细胞与不同NTs对背根神经节(DRG)感觉神经元中前速激肽原(PPT)、降钙素基因相关肽(CGRP)、神经丝200(NF - 200)和微管相关蛋白2(MAP - 2)mRNA表达的关联,我们所知甚少。在本研究中,建立了背根神经节(DRG)神经元与SKM细胞解离后的神经肌肉共培养模型。用倒置相差显微镜观察共培养中DRG神经元和SKM细胞的形态。通过实时PCR分析神经生长因子(NGF)或神经营养因子 - 3(NT - 3)对PPT、CGRP、NF - 200和MAP - 2 mRNA表达的影响。不同条件下神经肌肉共培养中DRG神经元细胞体和SKM细胞的形态相似。在神经肌肉共培养中存在不同NTs的情况下,神经元呈现出密集神经突生长的迹象。在神经肌肉共培养中,NGF和NT - 3增加了PPT、CGRP和NF - 200的mRNA水平,但未增加MAP - 2的mRNA水平。这些结果为更好地理解靶标SKM细胞与不同NTs对PPT、CGRP、NF - 200和MAP - 2 mRNA表达的关联提供了新线索,并暗示了靶标SKM细胞和NTs与DRG感觉神经元表型的关联。