Search

DNAX activating protein of 12 kDa (DAP12)-deficiency mice showed impaired differentiation of oligodendrocytes and reduced myelin in the central nervous system. Whether DAP12 is expressed by Schwann cells and its roles in the peripheral nervous system (PNS) remains unknown. In this study, expression of DAP12 was detected in Schwann cells in vivo and in vitro. The DAP12-knockout (KO) mice showed age-related motor deficits and thinner myelin in the sciatic nerve than WT mice but significantly faster clinical recovery after sciatic nerve crush injury. In sciatic nerves of DAP12 KO and WT mice, proteomic profiles analysis identified 158 differentially expressed proteins (DEPs) at 8-week-old, 29 DEPs at 54-week-old and 33 DEPs at two weeks after crush injury. Typically, of the DEPs at 54-week-old, up-regulated Lgmn and down-regulated RecK and Yap1 were associated with myelin loss in the sciatic nerve of DAP12 KO mice. Upregulation of nicotinamide nucleotide transhydrogenase and haptoglobin were associated with the accumulation of macrophages in the crushed sciatic nerve of DAP12 KO mice. After crush injury, there were significantly more M1 macrophages at one-week and more M2 macrophages at two-week in sciatic nerve of DAP12 KO mice than WT mice, indicating that DAP12 deletion promotes the phenotype conversion of macrophages from M1 to M2. Collectively, our findings suggest that DAP12 may exert dual roles in the PNS including promoting the physiological myelin formation and maintenance of Schwann cells but delaying nerve repair after injury by modulating the recruitment of macrophages and phenotype conversion. [Display omitted] • We demonstrated the expression of DNAX activating protein of 12 kDa (DAP12) in Schwann cells. • DAP12 deletion causes age-related motor deficits in peripheral nerve. • DAP12 deletion promotes the clinical recovery of mutant mice from sciatic nerve crush injury. [ABSTRACT FROM AUTHOR]

Schwann cells (SCs) isolation is one of the basic techniques for study of peripheral nervous system and peripheral neuropathy. A combined and effective method of isolating SCs from sciatic nerves of newborn mice with high yield and purity is still lacking. Sciatic nerves from neonatal mice aged 3–5 days serve as the source of SCs. Removal of adjacent connective tissue and epineurium, treatment with arabinoside hydrochloride and differential cell detachment technique were applied to eliminate fibroblast contamination and increase the purity of SCs. Combined use of collagenase/dispase and trypsin was chosen to increase the yield of SCs. Culture dishes precoated with poly-l-lysine and laminin, culture medium supplemented with heregulin β-1 and forskolin, and reasonable cell seeding density were implemented to increase the growth and proliferation of cultured SCs. Immunostaining of S100β and p75 neurotrophin receptor was used to identify the purity of SCs. Our method is able to obtain high-yield SCs with a purity of 90% within five days and a purity more than 99% within seven days from sciatic nerves of neonatal mice. Previous SCs isolation mostly focused on rats or adult mice and have a few limitations due to fibroblasts contamination, low yield and time-consuming. Our method permits SCs isolation from neonatal mice with a high yield and purity of primary SCs within 7 days. We described a fast, efficient and step-by-step method of isolating SCs from sciatic nerves of neonatal mice with high yield and purity. [Display omitted] • A new method of isolating Schwann cells (SCs) from newborn mice was described. • A high yield and purity (> 99%) of SCs isolation were obtained. • The whole procedure can be completely accomplished within 7 days. [ABSTRACT FROM AUTHOR]