Previous animal studies have revealed significant involvement of genetics in nicotine intake; however, the extent of the genetic contribution to this behavior has not been well addressed. We report the first study of nine generations of selection for high and low voluntary nicotine intake in outbred Sprague-Dawley rats. Bidirectional mass selection resulted in progressively greater nicotine consumption in the high nicotine-preferring line but no decrease in nicotine intake in the low nicotine-preferring line across generations. Our estimated realized heritability for high voluntary nicotine intake is 0.26 vs close to zero for low voluntary nicotine intake. In contrast, we found no differences between the lines across generations for saccharine intake. These selected lines may provide useful animal models for identifying susceptibility and resistance genes and variants for controlling voluntary nicotine intake in rodents, although we recognize that more generations of selection of these two lines and independent replication of our selection for high and low nicotine-preferring lines are needed.