Effects of pre- and neonatal nicotine exposure in rodents: inconsistent evidence.

This review presents an analysis of the literature on behavioral effects of developmental exposure to nicotine, as assessed in rodent models that mimic the consequences for human offspring of maternal cigarette smoking. Despite the frequency of reports of low birth weight, hyperactivity, cognitive deficits, and psychiatric problems, inconsistencies exist in both the clinical and experimental literature. Confounding socioeconomic and other demographic variables may account for discrepancies in clinical reports, and the choice of developmental exposure period and the method of nicotine administration may explain differences in experimental outcomes. Analysis of a number of variables (e.g., physical, behavioral, and cognitive) shows that fetal exposure to nicotine does not consistently cause growth retardation or decreased birth weight, nor reliably affect motor activity. But combined pre- and neonatal exposure is likely to result in delayed reflex development, global impairments in learning and memory, and an increased incidence of symptoms that model psychiatric illness. There is also support for increased self-administration of nicotine and other drugs of abuse in animals exposed developmentally to nicotine, as well as potent effects on offspring responses to drug challenges. Unlike reports in the clinical literature, sexually dimorphic effects were not evident in most animal models. Possible neuroanatomical and cholinergic mechanisms responsible for behavioral changes are briefly discussed. Statistical and design considerations are provided to increase the translational value of this research and, most importantly, enhance the replicability of reported findings.

Statistical issues and techniques appropriate for developmental neurotoxicity testing: a report from the ILSI Research Foundation/Risk Science Institute expert working group on neurodevelopmental endpoints.

The data from developmental neurotoxicity (DNT) guideline studies present a number of challenges for statistical design and analysis. The importance of specifying the planned statistical analyses a priori cannot be overestimated. A review of datasets submitted to the US Environmental Protection Agency revealed several inadequate approaches, including issues of Type I error control, power considerations, and ignoring gender, time, and litter allocation as factors in the analyses. Since DNT studies include numerous experimental procedures conducted on the dam and offspring at several ages, it is not unusual to have hundreds of significance tests if each was analyzed separately. Two general approaches to control experiment-wise Type I inflation are: 1) statistical/design considerations that reduce the number of p-values, including factorial designs, multivariate techniques, and repeated-measures analyses; and 2) adjustments to the alpha level, including newer approaches that are less conservative than, for example, Bonferroni corrections. The design of the DNT study includes testing of both sexes, and gender must be included in the statistical analysis for the determination of sex-related differences, and, indeed, including both sexes may increase power. The influence of litter must be taken into account in the allocation of test animals as well as the statistical analyses. This manuscript reviews many key considerations in the analysis of DNT studies with recommendations for statistical approaches and reporting of the data.

Prenatal ethanol preferentially enhances reactivity of the dopamine D1 but not D2 or D3 receptors in offspring.

Reports of prenatal ethanol (ETOH) effects on the dopamine system are inconsistent. In an attempt to clarify this issue, dams were given 35% ethanol-derived calories as the sole nutrient source in a liquid diet from the 10th through the 20th day of gestation (ETOH). Controls were pair-fed (PF) an isocaloric liquid diet or given ad libitum access to laboratory chow (LC). Prenatal exposure to both liquid diets reduced body weight of offspring relative to LC controls, more so for ETOH than for PF exposure. Prenatal ETOH also decreased litter size and viability, relative to both LC and PF control groups. On postnatal days 21-23, male and female offspring were given an injection of saline vehicle or one of eight specific dopamine receptor agonists or antagonists. Immediately after injection subjects were placed in individual observation cages, and over the following 30 min, eight behaviors (square entries, grooming, rearing, circling, sniffing, yawning, head and oral movements) were observed and quantified. No prenatal treatment effects on drug-induced behaviors were observed for dopamine D2 (Apomorphine, DPAT or Quinpirole) or D3 (PD 152255, Nafadotride, Apo or Quin effects on yawning) receptor agonists or antagonists, or for the vehicle control. In contrast, prenatal treatment effects were seen with drugs affecting the dopamine D1 receptor. Both D1 agonists (SKF 38393) and antagonists (SCH 23390 and high doses of spiperone) altered behaviors, especially oral and sniffing behaviors, in a manner which suggested enhanced dopamine D1 drug sensitivity in both ETOH and PF offspring relative to LC controls. These results suggest that at this age, both sexes experience a prenatal undernutrition-linked increase in the behavioral response to dopamine D1 agonists and antagonists, which can be intensified by gestational exposure to alcohol.

Behavioral response profiles following drug challenge with dopamine receptor subtype agonists and antagonists in developing rat.

As part of an investigation into the effects of gestational ethanol (ETOH) exposure on the developing dopamine (DA) system, pregnant Sprague-Dawley rats were exposed to one of three conditions: ETOH, pair-fed (PF) to the ETOH group, or ad libitum lab chow controls (LC). In this paper we report behavioral drug challenge effects for offspring of the two control groups (PF and LC). Male and female pups between postnatal days (PNDs) 21 and 23 in age were exposed to one of three intraperitoneal/subcutaneous doses of one of eight drugs chosen to assess the functional status of the DA D(1), D(2), and D(3) receptor subtype, or a saline control. Agonists were SKF 38393, apomorphine (APO), quinpirole (QUIN), and 7-hydroxy-N,N-di-n-propyl-2-amino-tetralin [7-OH-DPAT (DPAT)]; antagonists were spiperone (SPIP), SCH 23390, and two recently developed D(3) antagonists nafadotride (NAF) and PD 152255. Immediately following drug injection, pups were placed in observation cages, where eight behaviors (square entries, grooming, circling, rearing, sniffing, head and oral movements, and yawning) were scored at 3-min intervals for 30 min. Classic behavioral profiles were generally obtained for the high-dose mixed agonists APO, DPAT, and QUIN, which potently increased square entries, rearing, and sniffing, while reducing grooming and head movements. However, low-dose APO had no effect on behavior. The D(1) agonist, SKF 38393, had a strikingly different behavioral profile; it had no effect on square entries at any dose, while increasing grooming and sniffing at the medium dose. The D(1) antagonist, SCH 23390, profoundly decreased all behaviors except oral and head movements, especially at high doses. In contrast, the effects of the D(2) antagonist, SPIP, were limited to increasing sniffing at the medium dose. The two putative D(3) antagonists, NAF and PD 152255, presented strikingly different profiles. NAF induced a pattern of behavioral suppression that resembled the profile of high-dose SCH, while high-dose PD 152255 stimulated behavior. The failure of low-dose APO to have any effect on behavior suggests that the D(2) autoreceptor is not functional in preweanling rats. This hypothesis is further supported by the lack of behavioral suppression seen with low-dose QUIN and DPAT. Failure of NAF to produce behavioral activation at low doses and the stimulatory effects seen with PD 152255 suggests that either the D(3) autoreceptor, the postsynaptic D(3) receptor, or both are not fully functional at this age as well.