Animal studies were performed according to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health and China. The Experimental Animal Centre of Hubei Medical University provided C57BL/6 mice (male, 3–5 months) that met the criteria. The Institutional Animal Care and Use Committee of Hubei Medical University approved the animal protocols (Cat: 2019–111).
For the transduction of adult muscles, C57BL/6 male mice were anesthetized by using an isoflurane vaporizer maintained at 2% isoflurane and 1 L/m oxygen. Gastrocnemius and soleus (SL) muscles were exposed and injected with Ad-Ezrin (1 × 1010 pfu, two points, 50 μm/each) [15 (link)]. Muscles were removed 7 days after transfection, frozen in isopentane cooled in liquid nitrogen, and stored at − 80 °C.
Mutation and deletion of L-periaxin was associated with Charcot-Marie-Tooth (CMT) characterized by progressive muscle weakness and atrophy of distal extremities with sensory impairment through destroying the myelin sheath formed by Schwann cells. Interestingly, Ezrin inhibits the self-association of L-periaxin and participates in myelin sheath maintenance [4 (link), 5 (link)]. To confirm whether L-periaxin/Ezrin independence and interaction participate in CMT and muscular atrophy, a peroneal nerve injury model was prepared. Briefly, C57BL/6 male mice were anesthetized by using an isoflurane vaporizer maintained at 2% isoflurane and 1 L/m oxygen. Peroneal nerves were exposed and clamped for 15 min; subsequently, the gastrocnemius muscle (GA) was injected with Ad-Ezrin, Ad-Periaxin or Ad-shPeriaxin alone (1 × 1010 pfu, three points, 50 μm/each), and combined treatment with Ad-Ezrin (1 × 1010 pfu, three points, 50 μm/each) injection into the GA with Ad-shPeriaxin injection into the GA or Ad-Periaxin incubation within the injured peroneal nerves was incubated with Ad-Periaxin (1 × 1010 pfu, 50 μm/each) [15 (link)]. The sham and PNI groups within the peroneal nerves and GA were treated with equal amounts of normal saline. Muscles were removed 14 days after transfection, frozen in isopentane cooled in liquid nitrogen, and stored at − 80 °C. The myofiber types were measured through double fluorescence immunostaining of MyHC-I (NOQ, ab234431) and MyHC-II (My32, ab51263). Masson and hematoxylin-eosin staining were performed according to the manufacturer’s instructions. The successful establishment of the peroneal nerve injury model is shown in Additional file1 : Figure S1.
For the transduction of adult muscles, C57BL/6 male mice were anesthetized by using an isoflurane vaporizer maintained at 2% isoflurane and 1 L/m oxygen. Gastrocnemius and soleus (SL) muscles were exposed and injected with Ad-Ezrin (1 × 1010 pfu, two points, 50 μm/each) [15 (link)]. Muscles were removed 7 days after transfection, frozen in isopentane cooled in liquid nitrogen, and stored at − 80 °C.
Mutation and deletion of L-periaxin was associated with Charcot-Marie-Tooth (CMT) characterized by progressive muscle weakness and atrophy of distal extremities with sensory impairment through destroying the myelin sheath formed by Schwann cells. Interestingly, Ezrin inhibits the self-association of L-periaxin and participates in myelin sheath maintenance [4 (link), 5 (link)]. To confirm whether L-periaxin/Ezrin independence and interaction participate in CMT and muscular atrophy, a peroneal nerve injury model was prepared. Briefly, C57BL/6 male mice were anesthetized by using an isoflurane vaporizer maintained at 2% isoflurane and 1 L/m oxygen. Peroneal nerves were exposed and clamped for 15 min; subsequently, the gastrocnemius muscle (GA) was injected with Ad-Ezrin, Ad-Periaxin or Ad-shPeriaxin alone (1 × 1010 pfu, three points, 50 μm/each), and combined treatment with Ad-Ezrin (1 × 1010 pfu, three points, 50 μm/each) injection into the GA with Ad-shPeriaxin injection into the GA or Ad-Periaxin incubation within the injured peroneal nerves was incubated with Ad-Periaxin (1 × 1010 pfu, 50 μm/each) [15 (link)]. The sham and PNI groups within the peroneal nerves and GA were treated with equal amounts of normal saline. Muscles were removed 14 days after transfection, frozen in isopentane cooled in liquid nitrogen, and stored at − 80 °C. The myofiber types were measured through double fluorescence immunostaining of MyHC-I (NOQ, ab234431) and MyHC-II (My32, ab51263). Masson and hematoxylin-eosin staining were performed according to the manufacturer’s instructions. The successful establishment of the peroneal nerve injury model is shown in Additional file
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