Here, we report on the identification of Itga7-expressing muscle resident glial cells activated by loss of NMJ integrity. Gene expression analysis at bulk and single cell level revealed that these cells are distinct from Itga7-expressing muscle satellite cells. We show that a selective activation and expansion of Itga7-positive glial cells occurs in response to muscle nerve lesion. Upon activation, muscle glial-derived progenies expressed neurotrophic genes, including Ngfr, which enables their isolation by FACS. We show that activated muscle glial cells also expressed genes potentially implicated in ECM remodeling at NMJs. Among them, we observed Tenascin C (Tnc), which was highly expressed by muscle glial cells activated upon nerve injury, and preferentially localized to NMJ. Interestingly, we observed that while the activation of muscle glial cells by acute nerve injury was reversible, upon NMJ repair. By contrast, in a mouse model of Amyotrophic Lateral Sclerosis (ALS), in which NMJ degeneration is progressive, muscle glial cells steadily increased over the course of the disease; however, they exhibited an impaired neurotrophic activity, suggesting that pathogenic activation of glial cells may be implicated in ALS progression.