Nicotine alters nicotinic receptor subunit levels differently in developing Mammalian sympathetic neurons

The subunit specific expression of nicotinic acetylcholine receptors (nAChRs) undergo changes in the superior cervical sympathetic ganglion (SCG) of rat pups during neonatal growth. Since nAChRs play a significant role in sympathetic transmission, and the effect of nicotine on the expression of nAChR subunits in neurons of neonatal SCG is not known, we determined the effects of nicotine on receptor profiles using primary cultures of SCG neurons of 1-day-old rat pups. Neurons in culture were exposed to 1 and 10 microM of nicotine in the presence and absence of nerve growth factor (NGF). After 24 h, protein from the control and experimental neuron cultures was analyzed for the presence of nAChR containing alpha7 and alpha3 subunits using subunit specific antibodies. Exposure to 1 microM of nicotine marginally increased alpha7 subunits only in the absence of NGF. However it increased the level of alpha3 subunits significantly by 18% and 33.6% in the presence and absence of NGF, respectively. Exposure to 10 microM of nicotine did not alter the levels of either of the subunits. Interestingly, when the neurons were pre-exposed to alpha-bungarotoxin (antagonist of alpha7 nAChR), exposure to 10 microM of nicotine resulted in significant increases not only in alpha7 nAChR (25.5%) but also in alpha3 nAChR (32.2%). These results show that exposure to nicotine alters the nAChR levels differently in the neonatal sympathetic neurons in a subunit specific manner and suggest that the level of alpha7 as well as alpha3 nAChR is linked to the functional status of alpha7 nAChR in these neurons.