Neuronal Control of Localized Inflammation through Expressed Nicotinic Acetylcholine Receptors: A Study Carried Out in Mice

Abstract

Introduction: Although the local inflammatory reactions are known to be regulated through cholinergic anti-inflammatory pathways, the exact subtypes of nicotinic acetylcholine receptors involved in neuroimmune modulation are not well identified. Objectives: Immunohistochemical localisation of α 1 subunit of nicotinic acetylcholine receptors (α1 nAChR) in sites of localised inflammation induced by injecting turpentine to the hind limbs of Balb/C mice. Methods: Localised inflammation and subsequent development of sterile abscesses was induced by injecting sterile turpentine subcutaneously into thighs of Balb/C mice. Sterile saline was used in the control. Skin and muscle tissues of inflammatory sites were recovered from the animals after 48 hours and were stained with hematoxylin and eosin. Indirect immunohistochemistry was done using anti-a1nAChR as the primary antibody and biotinylated anti-rat lgG as the secondary antibody. Labeled streptavidin biotin (LSAB) technique was used with diaminobenzedene to detect the immunoreactivity (IR). Intensity of immunostaining was determined based upon a score of 0 - 3+ by qualitative computerised image analysis using FSX 100 Olympus microscope. Results: H and E stained slides showed polymorphonuclear leukocytes (PNL) infiltration at the abscess sites while the saline injected control tissue sections did not show PNL infiltration. A 2+ immunoreactivity (IR) of α 1nAChRs was visible at peripheral zones of sterile abscesses where PNL infiltrations were high while the central area with necrotic tissue did not show IR. A subcutaneous lymph node found within the inflammatory region expressed IR of a 1nAChR in its capsular sinuses, subcapsular sinuses and trabecular regions. Conclusions: The findings suggest the possible role of controlling localised inflammatory response by parasympathetic cholinergic nerves through α1nAChRs of inflammation sites.