Title Two-Photon-Excited Fluorescence Microscopy as a Tool to Investigate the Efficacy of Methylprednisolone in a Mouse Spinal Cord Injury Model
Authors Zhang, Yiling
Zhang, Lihai
Shen, Jing
Chen, Chao
Mao, Zhi
Li, Wei
Gan, Wen-Biao
Tang, Peifu
Affiliation Gen Hosp Chinese Peoples Liberat Army, Dept Orthopaed, Beijing 100853, Peoples R China.
Peking Univ, Shenzhen Grad Sch, Sch Chem Biol & Biotechnol, Shenzhen, Peoples R China.
NYU, Sch Med, Dept Physiol, Mol Neurobiol Program, New York, NY USA.
NYU, Sch Med, Skirball Inst, New York, NY USA.
Keywords spinal cord injury
methylprednisolone
in vivo imaging
2-photon microscopy
axons
microglia
IN-VIVO
MICROGLIAL RESPONSE
REGENERATION
AXONS
NEUROPROTECTION
RECOVERY
RATS
Issue Date 2014
Publisher spine
Citation SPINE.2014,39,(8),E493-E499.
Abstract Study Design. Basic imaging experiment. Objective. To explore the use of 2-photon-excited fluorescence (2PEF) microscopy to investigate the therapeutic effect of methylprednisolone (MP) in mice with spinal cord injury (SCI). Summary of Background Data. MP can alleviate secondary SCI through its anti-inflammatory effect; however, how MP regulates axonal dynamics in a compression SCI model is not well characterized. We used 2PEF microscopy to trace axonal dynamics in vivo during MP therapy. Methods. Two types of transgenic mice (weighing 23-25 g) including YFP-H line (n = 18) and CX3CR1-GFP (n = 18) were used for experimental procedure. Each type of mouse was randomly divided into 3 groups, and the sample size of every subgroup was 6. The sham groups including YFP-H line group (n = 6) and CX3CR1GFP group (n = 6) received laminectomy only (group 1). SCI groups received saline treatment (group 2) and SCI groups received MP treatment (group 3). Hind limb motor function was evaluated using the Basso Mouse Scale. 2PEF microscopy was used to image in vivo axonal dynamics at baseline and at 0.5 hours, 24 hours, 48 hours, and 72 hours postinjury. Histology was employed to examine pathological changes and microglial/macrophage proliferation after all imaging sessions. Results. Group 1 exhibited no significant differences in hind limb motor function before versus after surgery. The Basso Mouse Scale scores were significantly lower in groups 2 and 3 than in group 1 (P < 0.05). Degree of recovery was higher in group 3 than in group 2 at 7 days postinjury (P < 0.05). The axons in group 1 remained intact at all time points. The survival rate of axons in groups 2 and 3 progressively decreased at 48 hours postinjury; at 72 hours postinjury, the axon survival rate was higher in group 3 than group 2 (P < 0.05). Histology revealed that group 3 presented milder damage in injured spinal cord than group 2. Microglial/macrophage proliferation was lower in group 3 than in group 2 (P < 0.05). Conclusion. 2PEF microscopy is useful for detecting early changes, indicating axonal disruption in compression SCI. MP therapy may help alleviate axonal progressive damage and reduce the proliferation of microglia/macrophages in acute SCI.
URI http://hdl.handle.net/20.500.11897/342721
ISSN 0362-2436
DOI 10.1097/BRS.0000000000000218
Indexed SCI(E)
PubMed
Appears in Collections: 化学生物学与生物技术学院

Files in This Work
There are no files associated with this item.

Web of Science®


7

Checked on Last Week

Scopus®



Checked on Current Time

百度学术™


0

Checked on Current Time

Google Scholar™





License: See PKU IR operational policies.