Niemann-Pick Type C1 deficiency in microglia does not cause neuron death in vitro

Niemann-Pick Type C (NPC) disease is an autosomal recessive disorder that results in accumulation of cholesterol and other lipids in late endosomes/lysosomes and leads to progressive neurodegeneration and premature death. The mechanism by which lipid accumulation causes neurodegeneration remains unclear. Inappropriate activation of microglia, the resident immune cells of the central nervous system, has been implicated in several neurodegenerative disorders including NPC disease. Immunohistochemical analysis demonstrates that NPC1 deficiency in mouse brains alters microglial morphology and increases the number of microglia. In primary cultures of microglia from Npc1−/− mice cholesterol is sequestered intracellularly, as occurs in other NPC-deficient cells. Activated microglia secrete potentially neurotoxic molecules such as tumor necrosis factor-α (TNFα). However, NPC1 deficiency in isolated microglia did not increase TNFα mRNA or TNFα secretion in vitro. In addition, qPCR analysis shows that expression of pro-inflammatory and oxidative stress genes is the same in Npc1+/+ and Npc1−/− microglia, whereas the mRNA encoding the anti-inflammatory cytokine, interleukin-10 in Npc1−/− microglia is ~ 60% lower than in Npc1+/+ microglia. The survival of cultured neurons was not impaired by NPC1 deficiency, nor was death of Npc1−/− and Npc1+/+ neurons in microglia–neuron co-cultures increased by NPC1 deficiency in microglia. However, a high concentration of Npc1−/− microglia appeared to promote neuron survival. Thus, although microglia exhibit an active morphology in NPC1-deficient brains, lack of NPC1 in microglia does not promote neuron death in vitro in microglia–neuron co-cultures, supporting the view that microglial NPC1 deficiency is not the primary cause of neuron death in NPC disease.

► Microglia are activated and proliferate in NPC1-deficient mouse brains.
► Tumor necrosis factor mRNA and secretion are not increased in Npc1−/− microglia.
► Survival of cultured neurons was not compromised by NPC1 deficiency.
► Npc1−/− microglia do not increase neuron death in microglia–neuron co-cultures.