Cocaine potently blocks neuronal α3β4 nicotinic acetylcholine receptors in SH-SY5Y cells

• Published in: Acta pharmacologica Sinica 2020-02, Vol.41 (2), p.163-172
• Main Authors: Ma, Ze-gang, Jiang, Nan, Huang, Yuan-bing, Ma, Xiao-kuang, Brek Eaton, Jason, Gao, Ming, Chang, Yong-chang, Lukas, Ronald J, Whiteaker, Paul, Neisewander, Janet, Wu, Jie
• Format: Article
• Language: English
• Subjects: Acetylcholine receptors (nicotinic); Biomedical and Life Sciences; Biomedicine; Cocaine; Dopamine transporter; Drug abuse; Immunology; Internal Medicine; Internalization; Intracellular; Medical Microbiology; Nicotine; Perfusion; Pharmacology/Toxicology; Vaccine
• ISSN: 1671-4083; 1745-7254
• DOI: 10.1038/s41401-019-0276-y

Cocaine is one of the most abused illicit drugs worldwide. It is well known that the dopamine (DA) transporter is its major target; but cocaine also acts on other targets including nicotinic acetylcholine receptors (nAChRs). In this study, we investigated the effects of cocaine on a special subtype of neuronal nAChR, α 3 β 4 -nAChR expressed in native SH-SY5Y cells. α 3 β 4 -nAChR-mediated currents were recorded using whole-cell recordings. Drugs were applied using a computer-controlled U-tube drug perfusion system. We showed that bath application of nicotine induced inward currents in a concentration-dependent manner with an EC 50 value of 20 µM. Pre-treatment with cocaine concentration-dependently inhibited nicotine-induced current with an IC 50 of 1.5 μM. Kinetic analysis showed that cocaine accelerated α 3 β 4 -nAChR desensitization, which caused a reduction of the amplitude of nicotine-induced currents. Co-application of nicotine and cocaine (1.5 μM) depressed the maximum response on the nicotine concentration-response curve without changing the EC 50 value, suggesting a non-competitive mechanism. The cocaine-induced inhibition of nicotine response exhibited both voltage- and use-dependence, suggesting an open-channel blocking mechanism. Furthermore, intracellular application of GDP-βS (via recording electrode) did not affect cocaine-induced inhibition, suggesting that cocaine did not alter receptor internalization. Moreover, intracellular application of cocaine (30 µM) failed to alter the nicotine response. Finally, cocaine (1.5 μM) was unable to inhibit the nicotine-induced inward current in heterologous expressed α 6 /α 3 β 2 β 3 -nAChRs and α 4 β 2 -nAChRs expressed in human SH-EP1 cells. Collectively, our results suggest that cocaine is a potent blocker for native α 3 β 4 -nAChRs expressed in SH-SY5Y cells.