Denys deCatanzaro

Behavioral and molecular changes in the mouse in response to prenatal exposure to the anti-epileptic drug valproic acid

Experiments in rodents have indicated that maternal valproic acid (VPA) exposure has permanent adverse effects upon neurological and behavioral development. In humans, prenatal exposure to VPA can induce fetal valproate syndrome, which has been associated with autism. The present study examined mouse pups exposed in utero to VPA, measuring physical development, olfactory discrimination, and social behavior as well as expression of plasticity-related genes, brain derived neurotrophic factor (BDNF) and NMDA receptor subunits NR2A and NR2B. VPA-exposed mice showed delayed physical development, impaired olfactory discrimination, and dysfunctional pre-weaning social behavior. In situ hybridization experiments revealed lower cortical expression of BDNF mRNA in VPA animals. These results support the validity of the VPA mouse model for human autism and suggest that alterations in plasticity-related genes may contribute to the behavioral phenotype.

Human suicide: a biological perspective

Human suicide presents a fundamental problem for the scientific analysis of behavior. This problem has been neither appreciated nor confronted by research and theory. Almost all other behavior exhibited by humans and nonhumans can be viewed as supporting the behaving organisms biological fitness and advancing the welfare of its genes. Yet suicide acts against these ends, and does so more directly and unequivocally than any other form of maladaptive behavior. Four heuristic models are presented here to account for suicide in an evolutionary and sociobiological framework. The first model attributes suicide to the extraordinary development of learning and cultural evolution in the human species. Learning may make human behavior so independent of biological constraints that it can occasionally assume a form entirely contrary to the principles of biological evolution. The second model attributes suicide to a breakdown of adaptive mechanisms in extremely stressful novel environments. The third model involves kin and group selection, arguing that in limited circumstances suicide may occur because of beneficial effects it has on other, surviving individuals who share the suicidal individuals genes. The last model suggests that suicide should be tolerated by evolution when it has no effect on the gene pool. This model holds particular promise in accounting for aspects of suicide not attributable to culture. The evidence indicates that suicide is most common in individuals who are unlikely to reproduce and unable to engage in productive activity; such individuals are least capable of promoting their genes. A complete explanation of suicide may derive only from an analysis of its biological significance.

Bisphenol-A exposure during the period of blastocyst implantation alters uterine morphology and perturbs measures of estrogen and progesterone receptor expression in mice.

Bisphenol-A (BPA) has estrogenic properties both in vitro and in vivo. We investigated its impacts upon uterine morphology and estrogen and progesterone receptors after injection on gestational days 1-4 in doses known to disrupt pregnancy. Blastocyst implantation was significantly reduced by doses of 6.75 and 10.125 mg/animal. Uterine luminal area expanded substantially in response to increasing doses of BPA. Luminal epithelial cell height increased following exposure to 10.125 mg/animal, whereas there were no differences in the number of corpora lutea among conditions. The proportion of cells staining positively for estrogen receptors was affected non-monotonically, showing highest levels at 3.375 mg/animal and lowest levels at 10.125 mg/animal. Similarly progesterone receptor expression measured through western blots related non-monotonically to dose, being highest at 3.375 mg/animal and diminishing with increasing dose. These results suggest that BPA exposure during early gestation acts at the uterus to disrupt intrauterine implantation, consistent with an estrogenic effect.

Impact of acute bisphenol-A exposure upon intrauterine implantation of fertilized ova and urinary levels of progesterone and 17β-estradiol

Bisphenol-A (BPA), a monomer used in production of polycarbonate plastics and epoxy resins, has established estrogenic properties. We assessed the impact of acute and repeated subcutaneous BPA administration upon intrauterine implantation of fertilized ova and urinary levels of 17beta-estradiol and progesterone in inseminated female mice. In Experiment 1, females received varied doses of BPA on days 1-4 of gestation. Daily doses of 6.75 and 10.125mg/animal significantly reduced the number of implantation sites. Urinary progesterone was significantly reduced by the higher dose, but no other dose had an effect on progesterone levels and no dose altered estradiol levels. In Experiment 2, inseminated females received a single dose of BPA on days 0, 1, or 2 of gestation. A single dose of 10.125mg reduced the number of implantation sites when given on day 0 or day 1, and 6.75mg on day 1 also produced fewer implantation sites, but there was no such effect of any dose when administered on day 2. These data show a lower threshold for BPA-induced pregnancy disruption than previously reported, also indicating effects of just one exposure. They confirm that this disruption is due to the actions of BPA upon implantation sites, and show that higher doses can influence systemic progesterone levels.

Prenatal exposure to valproic acid leads to reduced expression of synaptic adhesion molecule neuroligin 3 in mice.

In rodents, a single administration of valproic acid (VPA) in utero leads to developmental delays and lifelong deficits in motor performance, social behavior, and anxiety-like behavior in the offspring. Recently, we have demonstrated that VPA mice show alterations in postnatal growth and development, and deficits in olfactory discrimination and social behavior early in development. Based on behavioral and molecular parallels between VPA rodents and individuals with autism, maternal challenge with VPA has been suggested to be a good animal model of autism. Neuroligins (NLGN) are a family of postsynaptic cell-adhesion molecules that play a role in synaptic maturation through association with their presynaptic partners, the neurexins (NRXN). Both NLGNs and NRXN members have been implicated in genetic studies of autism. In the present study, we examined changes at the level of expression of NLGN and NRXN mRNAs in the adult brain from mice exposed in utero to VPA. Mouse brain tissue was processed using in situ hybridization and analyzed with densitometry to examine expression of three NLGN genes (NLGN1, NLGN2, and NLGN3) and three NRXN genes (NRXN1, NRXN2, and NRXN3). Expression levels of NLGN1, NLGN2, NRXN1, NRXN2, and NRXN3 were observed to be similar in VPA and control mice. NLGN3 mRNA expression was found to be significantly lower in the VPA mice relative to control animals in hippocampal subregions, cornu ammonis (CA1) and dentate gyrus, and somatosensory cortex. This lowered expression may be linked to autistic-like behavioral phenotype observed in the VPA mice.

Responsiveness to laboratory pain in women as a function of age and childbirth pain experience

&NA; Pain responsiveness was investigated experimentally as a function of age and childbirth pain experience. Sensitivity to cold pressor‐induced pain was assessed through threshold, tolerance, and visual analog pain ratings. It was hypothesized that childbirth pain experience would mostly modify experimental pain judgment, in accordance with the adaptation‐levels model. That is, childbirth pain would be used as an “anchor” in evaluating other painful events. Fifteen parous women were compared to 12 nulliparous women of the same age (mean age: 35 years) as well as to 15 nulliparous younger women (mean age: 24 years). This comparison was undertaken in order to distinguish the effects of age, which was found to correlate with pain threshold. Analysis of variance comparing the three groups of women was performed on each of the three cold pressor measures. A significant effect was found for pain threshold. Multiple comparisons indicated that parous women had a higher pain threshold than both groups of nulliparous women which did not differ from one another. Thus, painful childbirth experience is sufficient to raise cold pressor pain threshold. This finding has never before been reported in the pain literature. It is consistent with anecdotal reports from parous women who, when providing cold pressor pain judgments, say that “nothing compares to labor pain.”

Estrogenicity of parabens revisited: impact of parabens on early pregnancy and an uterotrophic assay in mice.

Parabens, a class of preservatives routinely added to cosmetics, pharmaceuticals, and foods, have estrogenic properties. Given that intrauterine implantation of fertilized ova in inseminated females can be disrupted by minute levels of exogenous estrogens, we assessed the impact of parabens upon early gestation. In Experiment 1, butylparaben was administered subcutaneously in several doses ranging from 0.05 to 35 mg/animal/day to inseminated CF-1 mice on days 1-4 of pregnancy. Butylparaben exposure did not affect litter size, the number of pups born, postnatal day 3 litter weights, or the number of pups surviving to postnatal day 5. In contrast, administration of 500 ng/animal/day 17beta-estradiol terminated all pregnancies. In Experiment 2, propylparaben was subcutaneously administered to inseminated CF-1 mice on gestational days 1-4. Dams were sacrificed on gestation day 6 and the number of implantation sites was counted. Propylparaben had no impact on the number of implantation sites observed. Since Experiments 1 and 2 did not yield the anticipated results, an uterotrophic assay was conducted in Experiment 3 to re-evaluate the in vivo estrogenicity of parabens. Ovariectomized CF-1 and CD-1 mice were administered butylparaben in doses ranging from 0.735 to 35 mg per animal for three consecutive days. Mice were sacrificed on the fourth day, and uterine mass was recorded. There was no effect of butylparaben on uterine wet or dry mass at any dose in either strain. In contrast, administration of 17beta-estradiol consistently increased uterine mass in both strains. These data indicate that the estrogen-sensitive period of implantation is not vulnerable to paraben exposure, and that the in vivo estrogenicity of parabens may not be as potent as previously reported.

Chronic stress increases estrogen and other steroids in inseminated rats

Physical restraint, like many other stressors, can block early pregnancy, but the underlying physiological mechanisms have not been established. Exogenous estrogens in minute doses will also block early pregnancy. In the present study, female rats were exposed to 5 h of restraint daily for the first 5 days after insemination. A subset of animals was sacrificed after 1, 2, 3, 4, and 5 days of restraint, and blood was collected for radioimmunoassay. Blood was also collected from unstressed control animals on each of the first 5 days after insemination. Plasma concentrations of estradiol, as well as those of corticosterone and progesterone, were increased in the stressed animals as compared to the controls. The finding of significantly enhanced maternal estrogen suggests the possibility that estrogens mediate psychogenic pregnancy blocks.

Disruption of blastocyst implantation by triclosan in mice: impacts of repeated and acute doses and combination with bisphenol-A.

Triclosan is an antimicrobial additive in many personal care and household products, and evidence indicates that it can be estrogenic. As estrogen elevations can disrupt blastocyst implantation, we examined the influence of triclosan on implantation in inseminated mice. Doses of 18 and 27 mg/animal/day (about 523 and 785 mg/kg/day) on gestational days (GD) 1-3 reduced the number of implantation sites on GD 6. Single doses on GD 2 or 3 also reduced implantation sites. Subsequently, we examined triclosan in combination with bisphenol-A (BPA), which also can disrupt implantation. Although doses of 4 mg BPA (122 mg/kg) and 9 mg triclosan (262 mg/kg) on GD 1-3 were individually ineffective, in combination they reduced the number of implantation sites and also increased gestation length. All of these effects mimicked stronger effects of 17β-estradiol. These data are consistent with potential estrogenic properties of triclosan, and show that it can act together with BPA.

Non-invasive repeated measurement of urinary progesterone, 17β-estradiol, and testosterone in developing, cycling, pregnant, and postpartum female mice

Excretory samples from adult female mice were collected non-invasively during development, estrous cycling, pregnancy, and postpartum. In initial studies, urinary measures were statistically more dynamic over days than were fecal measures; thus subsequent studies focused on urine. Higher 17beta-estradiol levels were present in isolated females than in those exposed to males. In cycling females, urinary 17beta-estradiol was more variable than were measures of testosterone or progesterone, showing peaks with an approximate 5-day periodicity. When urinary estradiol and progesterone were monitored in conjunction with vaginal smear cell counts, patterns were idiosyncratic; most females showed distinct peaks in urinary steroids, not in clear synchrony with vaginal cell cornification. Levels of progesterone rose markedly during the first 10 days of pregnancy, then declined before birth. Estradiol showed a substantial peak on days 7-8 of gestation in all females measured. Urinary testosterone was not dynamic during pregnancy, but rose in immediate prenatal and postpartum measures. During post-weaning, pre-pubertal development, urinary levels of progesterone remained constant but levels of estradiol rose substantially over time.