Tissue-dependent effects of immunization with a nicotine conjugate vaccine on the distribution of nicotine in rats.

Vaccination of rats against nicotine reduces nicotine distribution to brain even at nicotine doses greatly exceeding the estimated binding capacity of the available antibody. This observation suggests a differential effect by which vaccination reduces nicotine distribution to brain to a greater extent than to other tissues. To test this hypothesis, vaccinated rats received a single intravenous nicotine dose equal to twice the estimated binding capacity of nicotine-specific antibody in vaccinated rats. The total and bound serum nicotine concentrations were higher in the vaccinated rats compared to controls, while the unbound serum nicotine concentration was lower. Distribution of nicotine to brain was reduced in vaccinated rats in a time-dependent manner, with a greater reduction at 1 min (64%) than at 25 min (45%). Vaccination reduced nicotine distribution to muscle, testis, spleen, liver, heart, and kidney, but to a lesser extent than to brain, while nicotine distribution to fat was increased. Chronically infused nicotine showed a similarly altered pattern of tissue distribution in vaccinated rats, but differences were in general smaller than after a single nicotine dose; brain nicotine concentration was 24% lower in vaccinated rats, while lung nicotine concentration was higher. The presence of nicotine-specific antibody in tissues may have contributed to the increased nicotine concentrations in fat and lung. These data suggest that vaccination reduces nicotine distribution to brain not only by sequestering nicotine in serum but also by redirecting tissue distribution disproportionately away from brain, such that nicotine concentrations are reduced to a greater extent in brain than in other tissues.

Detecting subtle differences in behavior using waveform display analysis.

We have devised a method, behavioral waveform display analysis, to analyze complex ethological information by measuring behavior in real time and visualizing it as a time-dependent, multistate waveform. To facilitate the generation and statistical analysis of behavioral waveform displays, we have designed a simple Macintosh-based software program. When keystrokes coded to particular behavioral states are entered in real time, this software measures and collates the time, frequency, and duration of each behavioral state. These data can then be displayed either in a tabular format for statistical analysis of behavioral duration and frequency or as graphical coordinates for creating waveform displays by direct importing into graphing programs. An illustration of the use of waveform display analysis to detect anomalous behaviors in cocaine- and amphetamine-treated mice, some of which are not detectable by a standard time-sampling assay, is shown. Both waveform display and time-sampling analysis detected drug-induced changes in sniffing, bar hanging, digging, and rearing. However, unlike time-sampling analysis, waveform display analysis also detected changes in the total duration, frequency, and average duration of these behaviors as well as additional changes in gnawing and locomotion. Additionally, visual scanning of behavioral waveform displays detected drug-induced changes in the patterns of behavior not detectable by time-sampling, including 1) a staged progression to a limited behavioral repertoire consisting of sniffing, locomotion, and rearing; 2) rapid switching between these remaining few behaviors; 3) a delayed onset of postinjection rearing relative to sniffing and locomotion; and 4) the absence of other transient stereotypies during the onset of drug action. These data indicate that behavioral waveform display provides an approach for the detection, visualization, and statistical analysis of aspects of complex behavior not amenable to detection by time-sampling methods.