Nicotinic receptors on rat alveolar macrophages dampen ATP-induced increase in cytosolic calcium concentration

• Main Authors: Lohmeyer Jürgen, Kurzen Hjalmar, Pfeil Uwe, Lips Katrin S, Zasłona Zbigniew, Jositsch Gitte, Hartmann Petra, Mikulski Zbigniew, Clauss Wolfgang G, Grau Veronika, Fronius Martin, Kummer Wolfgang
• Format: Article
• Language: English
• Published: BMC 2010-09-01
• Series: Respiratory Research
• Online Access: http://respiratory-research.com/content/11/1/133
• ISSN: 1465-9921

<p>Abstract</p> <p>Background</p> <p>Nicotinic acetylcholine receptors (nAChR) have been identified on a variety of cells of the immune system and are generally considered to trigger anti-inflammatory events. In the present study, we determine the nAChR inventory of rat alveolar macrophages (AM), and investigate the cellular events evoked by stimulation with nicotine.</p> <p>Methods</p> <p>Rat AM were isolated freshly by bronchoalveolar lavage. The expression of nAChR subunits was analyzed by RT-PCR, immunohistochemistry, and Western blotting. To evaluate function of nAChR subunits, electrophysiological recordings and measurements of intracellular calcium concentration ([Ca²⁺]_i) were conducted.</p> <p>Results</p> <p>Positive RT-PCR results were obtained for nAChR subunits α3, α5, α9, α10, β1, and β2, with most stable expression being noted for subunits α9, α10, β1, and β2. Notably, mRNA coding for subunit α7 which is proposed to convey the nicotinic anti-inflammatory response of macrophages from other sources than the lung was not detected. RT-PCR data were supported by immunohistochemistry on AM isolated by lavage, as well as in lung tissue sections and by Western blotting. Neither whole-cell patch clamp recordings nor measurements of [Ca²⁺]_irevealed changes in membrane current in response to ACh and in [Ca²⁺]_iin response to nicotine, respectively. However, nicotine (100 μM), given 2 min prior to ATP, significantly reduced the ATP-induced rise in [Ca²⁺]_iby 30%. This effect was blocked by α-bungarotoxin and did not depend on the presence of extracellular calcium.</p> <p>Conclusions</p> <p>Rat AM are equipped with modulatory nAChR with properties distinct from ionotropic nAChR mediating synaptic transmission in the nervous system. Their stimulation with nicotine dampens ATP-induced Ca²⁺-release from intracellular stores. Thus, the present study identifies the first acute receptor-mediated nicotinic effect on AM with anti-inflammatory potential.</p>