Journal: PLoS ONE
Article Title: Sympathetic Neurotransmitters Modulate Osteoclastogenesis and Osteoclast Activity in the Context of Collagen-Induced Arthritis
doi: 10.1371/journal.pone.0139726
Figure Lengend Snippet: Adrenergic receptors α1D, α2B, β2; muscarinic ACh receptor M5 and the alternative VIP receptor PACAP receptor 1 were stained positive on osteoclasts derived from BMMs of controls and CIA animals 20 (A) and 40 (B) days post-immunization. Paraformaldehyde-fixed cells were stained after 5 days differentiation. Nuclei were counterstained with DAPI. Cells containing 3 or more nuclei are considered to be osteoclasts (white circles). N = 4 rats. Magnification 200x. ACh: acetylcholine, AR: adrenoceptor, BMM: bone marrow-derived macrophages, CIA: collagen-induced arthritis, mAChR: muscarinic ACh receptor, NA: noradrenaline, PACAP: pituitary adenylate cyclase-activating peptide, VIP: vasoactive intestinal peptide
Article Snippet: Neurotransmitter receptors were detected using: rabbit antibody against rat PACAP receptor 1 (1:50, sc–30018, Santa Cruz Biotechnologies, Dallas, TX, USA); rabbit antibody against muscarinic ACh receptor M5 (1:400, ab41171) and rabbit antibody against adrenoceptor β2 (3μg/ml, ab36956; both Abcam, Cambridge, UK), as well as rabbit antibodies against extracellular sections of adrenoceptors α1D (1:100, #AAR–019) and α2B (1:100, #AAR–021; Alomone Labs, Jerusalem, Israel).
Techniques: Staining, Derivative Assay