Nicotine, the primary addictive substance in tobacco, induces profound behavioral responses in mammals, but the underlying genetic mechanisms are not well understood. Caenorhabditis elegans exhibits behavioral responses to nicotine that parallel those observed in mammals, including acute response, tolerance, withdrawal, and sensitization. The worms have a simple, well-characterized nervous system, so they are well suited for investigating how specific neurotransmitters, receptors, and signaling molecules function within the context of the nervous system to produce behavior.

The aim of this study was to detect the proteins related to nicotine addiction in Caenorhabditis elegans. These proteins may be useful in revealing the molecular mechanism of nicotine addiction and in isolating the potential drug targets for nicotine addiction-associated disease therapy.

The experiments were carried out on C. elegans wild-type strain Bristol (N2). The worms were grown on S medium by using concentrated E. coli OP50 as a food source. The liquid cultures of C. elegans were treated with nicotine (30 mM) for 1 h. The animals were homogenized in 50 mM HEPES-KOH (pH 7.6), 1 mM EDTA, 140 mM KCl, 0.5% NP-40, and 10% glycerol. The accumulation of proteins was analyzed by 16-BAC/SDS-PAGE for resolution of integral membrane proteins (Hartinger et al. 1996 Analyt. Biochem. 240: 126-133). The first-dimension separation occurs according to molecular weight in an acidic discontinuous PAGE system (pH 4.1-2.1) using cationic benzyldimethyl-n-hexadecylammonium chloride (16-BAC) as a detergent. Because proteins show slightly different migration properties, as compared to the SDS-PAGE system, the resolution can be improved by subsequent second-dimension SDS-PAGE.

Nicotine induced changes in protein pattern. The most pronounced effect of acute exposure of naive animals to nicotine was the preferential accumulation of polypeptides with a molecular weight of 36 kDa. Further research has been undertaken to identify proteins sequences in selected spots by mass spectrometry.

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