Tracing Axonal Regeneration in Mice

Anterograde-tracing was performed to detect axonal regeneration as previously described [21 (link)]. Biotinylated dextran amine (BDA; MW,10000; Invitrogen) 10% w/v in sterile PBS was injected into two sites (0.4 μl/ site) of the spinal cord by micropipette. One week later, mice were anesthetized and perfused transcardially with 4% paraformaldehyde. Thirty-micron sections were incubated with Alexafluor 488-conjugated streptavidin (1:500; Invitrogen) for 1 h at room temperature. The images were acquired with a confocal microscope (Olympus, BX51).

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Li X., Yu Z., Zong W., Chen P., Li J., Wang M., Ding F., Xie M., Wang W, & Luo X. (2020). Deficiency of the microglial Hv1 proton channel attenuates neuronal pyroptosis and inhibits inflammatory reaction after spinal cord injury. Journal of Neuroinflammation, 17, 263.

Publication 2020

Biotinylated dextran amine (BDA; Alexafluor 488-conjugated streptavidin

Axonal Biotinylated dextran amine Confocal microscope Mice Paraformaldehyde Regeneration Spinal cord Sterile Streptavidin

Corresponding Organization :

Other organizations : Huazhong University of Science and Technology, Tongji Hospital

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Variable analysis

independent variables

Injection of biotinylated dextran amine (BDA; MW,10000; Invitrogen) 10% w/v in sterile PBS into two sites (0.4 μl/ site) of the spinal cord by micropipette

dependent variables

Axonal regeneration detected through anterograde-tracing

control variables

Mice were anesthetized
Mice were perfused transcardially with 4% paraformaldehyde
Thirty-micron sections were incubated with Alexafluor 488-conjugated streptavidin (1:500; Invitrogen) for 1 h at room temperature

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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