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With the implementation of the Food Quality Protection Act in 1996, more detailed evaluations of possible health effects of pesticides on developing organisms have been required. As a result, considerable developmental neurotoxicity (DNT) data have been generated on a variety of endpoints, including developmental changes in motor activity, auditory startle habituation, and various learning and memory parameters. One issue in interpreting these data is the level of variability for the measures used in these studies: excessive variability can obscure treatment-related effects, or conversely, small but statistically significant changes could be viewed as treatment related, when they might in fact be within the normal range. To aid laboratories in designing useful DNT studies for regulatory consideration, an operational framework for evaluating observed variability in study data has been developed. Elements of the framework suggest how an investigator might approach characterization of variability in the dataset; identification of appropriate datasets for comparison; evaluation of similarities and differences in variability between these datasets, and of possible sources of the variability, including those related to test conduct and test design. A case study using auditory startle habituation data is then presented, employing the elements of this proposed approach.

d-Methamphetamine (MA) is one of more than two dozen drugs included in the cytochrome P450-mediated "debrisoquine oxidation polymorphism" panel. The human gene (CYP2D6) is responsible for the "poor metabolizer" (PM) and "extensive metabolizer" (EM) phenotypes for drugs such as MA; a similar polymorphism (the CYP2D1 gene) exists in rats. Female Black or Dark Agouti rats exhibit the PM phenotype, whereas Sprague-Dawley (SD) rats show the EM trait. We sought to test the possibility that these strains of rats might exhibit altered MA-induced developmental neurotoxicity. Neonatal exposure to MA on days 11-20 has previously been shown to induce spatial learning deficits in Sprague-Dawley rats when tested as adults. Therefore, in the present experiment, on postpartum days 11 through 20, ACI (Black Agouti) and SD progeny were administered 30 mg/kg MA twice daily. MA treatment caused larger increases in mortality in ACI than in SD rats, suggesting that decreased MA metabolism leads to enhanced toxicity and lethality. Female offspring were assessed behaviorally as adults. No differences were observed in acoustic startle or straight swimming channel performance. In the Morris maze, both MA-treated rat strains showed longer latencies to find the hidden platform during acquisition, reinstatement, and shift trials, and spent less time in the target quadrant on probe trials; no strain differences in learning were found. Although these data do not support our hypothesis that MA-induced developmental neurotoxicity might be enhanced in the ACI rat, this interpretation is tempered by the high mortality rate (65%) of MA-treated ACI neonates, suggesting a possible "survivor effect" in this strain.

Neonatal exposure to methamphetamine (MA) has previously been shown to induce acoustic startle facilitation when the animals were tested as adults. The present experiment sought to replicate and extend this effect using a lower dose of MA and to determine if the effect varied as a function of prepulse stimulus intensity. Sprague-Dawley CD rat offspring were culled on the day after birth to eight (preferentially retaining females). On days 1-10, progeny were injected SC with either 20 mg/kg of d-MA twice per day (doses spaced at least 8 h apart) or distilled water. On postnatal day 50, offspring were administered 51 acoustic startle trials followed by 36 prepulse trials. Prepulse intensities were 0, 70, 75, 80, 85, or 90 dB. MA progeny showed augmented startle response amplitudes on both paradigms but the effect was most pronounced on the prepulse trials. Prepulse intensity interacted with MA treatment in that significant facilitation in the MA animals occurred on 0 and 70 dB prepulse trials but was only a trend (p < 0.10) on 75, 80, 85, and 90 dB trials. This implies that the effect of MA is most likely upon the basic startle reflex and not upon inhibitory pathways that modify startle reactivity.

Pregnant Sprague-Dawley rats were treated with 0, 5, 10, 15, or 20 mg/kg d-methamphetamine (MA), expressed as the free base, by SC injection (b.i.d., 8 h apart) on days 7-12 or 13-18 of gestation. Plasma concentration of MA and amphetamine were determined after the last dose. MA reduced gestation weight gain. The late exposure resulted in an increase in maternal and offspring mortality and reduced offspring growth. Offspring treated early in gestation with MA showed delayed development of early locomotion. In addition, memory impairment, evidenced by decreased target quadrant times and platform crossings on test trials and increased latency on reversal trials in the Morris spatial navigation maze, reduced spontaneous alternation, and lengthened passive avoidance retention latency was seen in the early treated high-dose groups. A reduction of serotonin was found in the nucleus accumbens following late exposure to MA at 20 mg/kg. Animals in both exposure groups had eye defects; however, the type of defect was dependent on the developmental stage at the time of dosing. Anophthalmia occurred only after early MA exposure, whereas folded retina was drug related only after late MA exposure. The behavioral effects did not show graded dose dependency; however, the effects were sensitive to exposure period. The early exposed animals had more alterations in behavior whereas the late exposed group showed higher mortality, reduced body weights, and neurochemical alterations.

Although the possible behavioral neurotoxic effects of in utero exposure to cocaine have been the subject of numerous experiments, only a limited number of different types of animal models of cocaine exposure, critical periods, or long-term effects of such exposures have been investigated. In the present experiment, the effects of multiple daily SC exposures to cocaine (20 mg/kg/dose x 5 doses per day) were investigated when administered to gravid Sprague-Dawley CD rats on embryonic days E7-12 or E13-18 compared to weight-matched, vehicle injected, pair-fed controls. Effects of exposure were assessed on general development, olfactory orientation behavior, early locomotion, startle reactivity, spontaneous motor activity, and learning on two different tasks (Morris and Cincinnati water mazes). The multiple cocaine dosing regimen produced maternal peak serum concentrations of cocaine 3 times higher than that of a single dose (approximately 1550 vs. approximately 550 ng/mL). Early-exposed cocaine offspring had lower olfactory orientation scores and reduced postweaning rearing and hole-poke motor activity, whereas late-exposed cocaine offspring had increased postweaning locomotor, rearing, and hole-poke activity. On the Morris hidden platform maze, the cocaine early-exposed females had longer latencies on acquisition than controls. On the Cincinnati multiple-T water maze, the early-exposed cocaine females and the late-exposed cocaine males had increased errors, whereas the early-exposed cocaine males had reduced errors. The effects on measures of learning, when taken together, and in light of their being in the early-exposed group, suggest that embryonic cocaine exposure may have subtle effects on cognition in the offspring as adults. Such effects represent a form of neurotoxicity not previously associated with prenatal cocaine exposure.