Effects of chemokine (C-C motif) ligand 1 on microglial function

Abstract

Microglia, which constitute the resident macrophages of the central nervous system (CNS), are generally considered as the primary immune cells in the brain and spinal cord. Microglial cells respond to various factors which are produced following nerve injury of multiple aetiologies and contribute to the development of neuronal disease. Chemokine (C-C motif) ligand 1 (CCL-1), a well-characterized chemokine secreted by activated T cells, has been shown to play an important role in neuropathic pain induced by nerve injury and is also produced in various cell types in the CNS, especially in dorsal root ganglia (DRG). However, the role of CCL-1 in the CNS and the effects on microglia remains unclear. Here we showed the multiple effects of CCL-1 on microglia. We first showed that CCR-8, a specific receptor for CCL-1, was expressed on primary cultured microglia, as well as on astrocytes and neurons, and was upregulated in the presence of CCL-1. CCL-1 at concentration of 1 ng/ml induced chemotaxis, increased motility at a higher concentration (100 ng/ml), and increased proliferation and phagocytosis of cultured microglia. CCL-1 also activated microglia morphologically, promoted mRNA levels for brain-derived neurotrophic factor (BDNF) and IL-6, and increased the release of nitrite from microglia. These indicate that CCL-1 has a role as a mediator in neuron-glia interaction, which may contribute to the development of neurological diseases, especially in neuropathic pain.

Keywords: 4′,6′-diamidino-2-phenylindole dihydrochloride; BDNF; CCL-1; CCL-2; CCL-21; CCR-8; CNS; CX3CL-1; Chemotaxis; Cytokines; DAPI; DMEM; DRG; FBS; GABA; GFAP; IL-1β; IL-6; Iba1; MAP2; Microglia; NMDA receptor 2B; NO; NR2B; PBS; Phagocytosis; Proliferation; RT-PCR; TCA-3; TNF-α; base pair; bp; brain-derived neurotrophic factor; cDNA; central nervous system; chemokine (CX3C motif) ligand 1; chemokine (C–C motif) ligand 1; chemokine (C–C motif) ligand 2; chemokine (C–C motif) ligand 21; chemokine (C–C motif) receptor 8; complementary DNA; dorsal root ganglia; dulbecco’s modified eagle medium; fetal bovine serum; glial fibrillary acid protein; interleukin 1β; interleukin 6; ionized calcium-binding adapter molecule 1; mRNA; messenger RNA; microtubule-associated protein 2; nitric oxide; phosphate buffered saline; reverse transcription polymerase chain reaction; thymus-derived chemotactic agent 3; tumour necrosis factor-α; γ-amino butyric acid.