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MiR-34a Regulates Axonal Growth of Dorsal Root Ganglia Neurons by Targeting FOXP2 and VAT1 in Postnatal and Adult Mouse

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Abstract

Hyperglycemia impairs nerve fibers of dorsal root ganglia (DRG) neurons, leading to diabetic peripheral neuropathy (DPN). However, the molecular mechanisms underlying DPN are not fully understood. Using a mouse model of type II diabetes (db/db mouse), we found that microRNA-34a (miR-34a) was over-expressed in DRG, sciatic nerve, and foot pad tissues of db/db mice. In vitro, high glucose significantly upregulated miR-34a in postnatal and adult DRG neurons, which was associated with inhibition of axonal growth. Overexpression and attenuation of miR-34a in postnatal and adult DRG neurons suppressed and promoted, respectively, axonal growth. Bioinformatic analysis suggested that miR-34a putatively targets forkhead box protein P2 (FOXP2) and vesicle amine transport 1 (VAT1), which were decreased in diabetic tissues and in cultured DRG neurons under high glucose conditions. Dual-luciferase assay showed that miR-34a downregulated FOXP2 and VAT1 expression by targeting their 3′ UTR. Gain-of- and loss-of-function analysis showed an inverse relation between augmentation of miR-34a and reduction of FOXP2 and VAT1 proteins in postnatal and adult DRG neurons. Knockdown of FOXP2 and VAT1 reduced axonal growth. Together, these findings suggest that miR-34a and its target genes of FOXP2 and VAT1 are involved in DRG neuron damage under hyperglycemia.

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Acknowledgements

This work was supported by NINDS grants R01 NS075084 (LW) and RO1 NS075156 (ZGZ) and NIDDK RO1 DK097519 (LW). We thank Dr. Paul Fernyhough and colleagues in his laboratory to train us how to culture adult DRG neurons.

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Authors and Affiliations

  1. Department of Neurology, Henry Ford Hospital, 2799 West Grand Boulevard, Detroit, MI, 48202, USA

    Longfei Jia, Michael Chopp, Lei Wang, Xuerong Lu, Yi Zhang, Alexandra Szalad & Zheng Gang Zhang

  2. Department of Neurolgoy, Xuanwu Hospital, Capital Medical University, Beijing, China

    Longfei Jia

  3. Department of Physics Oakland University, Rochester, MI, 48309, USA

    Michael Chopp

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  1. Longfei Jia
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Fig. s1

Physiological parameters of db/m and db/db mice at age of 20 weeks. a: Mouse body weight, blood glucose levels, and HbA1c levels of db/m and db/db mice. b-e: Neurological function measured by MCV (b), SCV (c), Von Frey test (d), tail flick test (e). f-g: Representative images (f) and their quantitative data (g) show intraepidermal nerve fiber (red) densities in plantar skin. These data show that db/db mice at age of 20 weeks exhibit diabetic peripheral neuropathy. n = 6 mice/group. * P < 0.05 vs db/m mice. (GIF 16 kb).

High Resolution Image (TIFF 495 kb).

Fig. s2

Comparison between SMI31 and TUJ1 staining. a: Representative microscopic images show the colocalization of SMI31 and TUJ1 staining of postnatal DRG neurons. Scale bar = 100 μm. (GIF 137 kb).

High Resolution Image (TIFF 2380 kb).

Fig. s3

Bioinformatic analysis of putative miR-34a target genes. a-d: Based on IPA and Targetscan analysis, putative miR-34a target genes were grouped to 4 pathways associated to nervous system, including development of neurons (a), neuritogenesis (b), branching of neuritis (c), and growth of neuritis (d). Genes with red color in each pathway were selected for Western blot analysis. (GIF 38 kb).

High Resolution Image (TIFF 515 kb).

Fig. s4

Levels of proteins in diabetic mouse tissues. a-c: Representative Western blot show protein levels of ADAM10, DCX, c-MET, NOTCH1, ROCK1, SYNJ1, and VAMP2 in DRG (a), sciatic nerve (SN, b) and foot pad (FP, c) tissues of db/db or db/m mice. Three individual mice were presented for each mouse group. β-actin bands in a-c of each group were shown in Fig. 3a, respectively. d-j: Quantitative data show levels of ADAM10 (d), DCX (e), c-MET (f), NOTCH1 (g), ROCK1 (h), SYNJ1 (i), and VAMP2 (j) in DRG, SN and FP. n = 6 mice/group. * P < 0.05 vs db/m mice. (GIF 65 kb).

High Resolution Image (TIFF 575 kb).

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Jia, L., Chopp, M., Wang, L. et al. MiR-34a Regulates Axonal Growth of Dorsal Root Ganglia Neurons by Targeting FOXP2 and VAT1 in Postnatal and Adult Mouse. Mol Neurobiol 55, 9089–9099 (2018). https://doi.org/10.1007/s12035-018-1047-3

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