In the developing and adult brain, neurotrophic growth factors support the growth and protec tion of dopaminergic neuronal systems. Recently, links between impaired neurotrophin support of dopamine (DA) neurons has be...In the developing and adult brain, neurotrophic growth factors support the growth and protec tion of dopaminergic neuronal systems. Recently, links between impaired neurotrophin support of dopamine (DA) neurons has been described in Parkinson’s Disease (PD). Fibro- blast growth factor (FGF) has a unique association with DA neurons in that FGF signaling is vitally important for the development and protection of adult DA neurons. We assessed the role of substantia nigra (SN)-expressed FGFs in the nigrostriatal dopaminergic system using a transgenic mouse, th-fgfr1(tk-). In these mice, generated by expression of dominant negative FGFR1(TK-) from the tyrosine hydroxylase (TH) gene promoter, reduced FGF signaling results in smaller and less dense adult nigrostriatal DA neurons, similar to what is observed in PD. With unilateral 6-hydroxydopamine (6-OHDA) lesions, th-fgfr1(tk-) mice exhibited extensive unilateral nigrostriatal damage with robust spontaneous (non-drug induced) asymmetrical turning and a decreased latency to remain on the accelerating rotarod. L-DOPA remains the gold standard for PD therapy despite debilitating hyperkinetic and dyskinetic side effects. The nicotinic acetylcholine system has recently been targeted as an alternative system to combat PD motor symptoms. Nicotine effectively stimulates dopaminergic transmission in the nigrostriatal pathway and mediates movement. Using unilaterally lesioned th-fgfr1(tk-) mice, long term (11 day) oral administration of nicotine increased spontaneous bidirectional turning and increased the latency before falling from the accelerating rotarod. In a separate analysis, L-DOPA treatment reversed directionality of rotation and further deepened motor discoordination, suggesting activation of hypersensitive postsynaptic DA receptors in the denervated striata. These results in a transgenic model of PD provide insights into the cellular mechanisms underlying L-DOPA and nicotinic therapies and offer further evidence of nicotine’s capacity to facilitate movement and enhance motor coordination in PD.展开更多
[Objectives]To characterize the expression pattern of Fibroblast growth factor 10(FGF10)during the differentiation of rat L6 myoblasts and to identify potential key transcription factors(TFs)regulating its expression ...[Objectives]To characterize the expression pattern of Fibroblast growth factor 10(FGF10)during the differentiation of rat L6 myoblasts and to identify potential key transcription factors(TFs)regulating its expression through bioinformatics approaches.[Methods]Rat L6 myoblasts were induced to differentiate by culturing them in DMEM supplemented with 2%donor horse serum(DHS).Morphological changes were observed using an inverted microscope.Cell samples were collected prior to induction(day 0)and on days 1,3,5,and 7 post-induction.The relative expression levels of FGF10 mRNA and protein at each time point were quantified using RT-qPCR and Western blot analysis,respectively.Furthermore,a 2000 bp sequence upstream of the transcription start site of the rat Fgf10 gene was extracted as the promoter region.Putative TF binding sites were predicted using four databases(TRANSFAC,JASPAR,HOCOMOCO,and CISBP),and high-confidence candidates were screened to construct a regulatory network.[Results]Morphological observations confirmed successful differentiation,as evidenced by the appearance of binucleated myotubes on day 3 and the formation of numerous thick,multinucleated myotubes by day 7.Both RT-qPCR and Western blot analysis demonstrated a significant dynamic expression pattern of FGF10.Expression levels were markedly upregulated during the early phase(days 1-3),reaching a peak on day 3(P<0.01),followed by a decline to basal levels during the late phase(days 5-7).Cross-validation across multiple databases identified 48 high-confidence TFs,among which Elf5,Tcf3,Nkx3-2,Zic2,Tcf7,and Egr1 were consistently predicted by all four databases.[Conclusions]FGF10 exhibits high expression levels during the early stage of differentiation,indicating its crucial role in the initiation of myogenesis.The six identified TFs serve as core candidate regulators of Fgf10 expression,offering novel insights into the molecular mechanisms underlying muscle development.展开更多
文摘In the developing and adult brain, neurotrophic growth factors support the growth and protec tion of dopaminergic neuronal systems. Recently, links between impaired neurotrophin support of dopamine (DA) neurons has been described in Parkinson’s Disease (PD). Fibro- blast growth factor (FGF) has a unique association with DA neurons in that FGF signaling is vitally important for the development and protection of adult DA neurons. We assessed the role of substantia nigra (SN)-expressed FGFs in the nigrostriatal dopaminergic system using a transgenic mouse, th-fgfr1(tk-). In these mice, generated by expression of dominant negative FGFR1(TK-) from the tyrosine hydroxylase (TH) gene promoter, reduced FGF signaling results in smaller and less dense adult nigrostriatal DA neurons, similar to what is observed in PD. With unilateral 6-hydroxydopamine (6-OHDA) lesions, th-fgfr1(tk-) mice exhibited extensive unilateral nigrostriatal damage with robust spontaneous (non-drug induced) asymmetrical turning and a decreased latency to remain on the accelerating rotarod. L-DOPA remains the gold standard for PD therapy despite debilitating hyperkinetic and dyskinetic side effects. The nicotinic acetylcholine system has recently been targeted as an alternative system to combat PD motor symptoms. Nicotine effectively stimulates dopaminergic transmission in the nigrostriatal pathway and mediates movement. Using unilaterally lesioned th-fgfr1(tk-) mice, long term (11 day) oral administration of nicotine increased spontaneous bidirectional turning and increased the latency before falling from the accelerating rotarod. In a separate analysis, L-DOPA treatment reversed directionality of rotation and further deepened motor discoordination, suggesting activation of hypersensitive postsynaptic DA receptors in the denervated striata. These results in a transgenic model of PD provide insights into the cellular mechanisms underlying L-DOPA and nicotinic therapies and offer further evidence of nicotine’s capacity to facilitate movement and enhance motor coordination in PD.
基金Supported by Guangdong Basic and Applied Basic Research Foundation(2023A1515110973)Guangdong Provincial Young Innovative Talents Project of General Colleges and Universities(2023KQNCX089)+6 种基金Key Field Special Project of Guangdong Provincial Colleges and Universities(2025ZDZX2077)the Rural Science and Technology Commissioner Program of Guangdong Province(KTP20240673)Undergraduate Higher Education Teaching Quality and Reform Projects of Guangdong Province(Yuejiao Gao Han[2024]No.9Yuejiao Gao Han[2024]No.30)Zhaoqing Science and Technology Innovation Guidance Project(Zhaoke[2025]No.4)Scientific Research Foundation of Zhaoqing University(QN202436)College Student Innovation Training Program Project(S202510580042).
文摘[Objectives]To characterize the expression pattern of Fibroblast growth factor 10(FGF10)during the differentiation of rat L6 myoblasts and to identify potential key transcription factors(TFs)regulating its expression through bioinformatics approaches.[Methods]Rat L6 myoblasts were induced to differentiate by culturing them in DMEM supplemented with 2%donor horse serum(DHS).Morphological changes were observed using an inverted microscope.Cell samples were collected prior to induction(day 0)and on days 1,3,5,and 7 post-induction.The relative expression levels of FGF10 mRNA and protein at each time point were quantified using RT-qPCR and Western blot analysis,respectively.Furthermore,a 2000 bp sequence upstream of the transcription start site of the rat Fgf10 gene was extracted as the promoter region.Putative TF binding sites were predicted using four databases(TRANSFAC,JASPAR,HOCOMOCO,and CISBP),and high-confidence candidates were screened to construct a regulatory network.[Results]Morphological observations confirmed successful differentiation,as evidenced by the appearance of binucleated myotubes on day 3 and the formation of numerous thick,multinucleated myotubes by day 7.Both RT-qPCR and Western blot analysis demonstrated a significant dynamic expression pattern of FGF10.Expression levels were markedly upregulated during the early phase(days 1-3),reaching a peak on day 3(P<0.01),followed by a decline to basal levels during the late phase(days 5-7).Cross-validation across multiple databases identified 48 high-confidence TFs,among which Elf5,Tcf3,Nkx3-2,Zic2,Tcf7,and Egr1 were consistently predicted by all four databases.[Conclusions]FGF10 exhibits high expression levels during the early stage of differentiation,indicating its crucial role in the initiation of myogenesis.The six identified TFs serve as core candidate regulators of Fgf10 expression,offering novel insights into the molecular mechanisms underlying muscle development.
