Developmental programming: Sex-specific programming of growth upon prenatal bisphenol A exposure.

In both human and animals, in utero exposure to bisphenol A (BPA), an endocrine-disrupting chemical used in the production of plastics and epoxy resins, has been shown to affect offspring reproductive and metabolic health during adult life. We hypothesized that the effect of prenatal exposure to environmentally relevant doses of BPA will be evident during fetal organogenesis and fetal/postnatal growth trajectory. Pregnant ewes were administered BPA subcutaneously from 30 to 90 days of gestation (term 147 days). Fetal organ weight, anthropometric measures, maternal/fetal hormones and postnatal growth trajectory were measured in both sexes. Gestational BPA administration resulted in higher accumulation in male than female fetuses only at fetal day 65, with minimal impact on fetal/maternal steroid milieu in both sexes at both time points. BPA-treated male fetuses were heavier than BPA-treated female fetuses at fetal day 90 whereas this sex difference was not evident in the control group. At the organ level, liver weight was reduced in prenatal BPA-treated female fetuses, while heart and thyroid gland weights were increased in BPA-treated male fetuses relative to their sex-matched control groups. Prenatal BPA treatment also altered the postnatal growth trajectory in a sex-specific manner. Males grew slower during the early postnatal period and caught up later. Females, in contrast, demonstrated the opposite growth trend. Prenatal BPA-induced changes in fetal organ differentiation and early life growth strongly implicate translational relevance of in utero contributions to reproductive and metabolic defects previously reported in adult female offspring.

Developmental Programming: Prenatal and Postnatal Androgen Antagonist and Insulin Sensitizer Interventions Prevent Advancement of Puberty and Improve LH Surge Dynamics in Prenatal Testosterone-Treated Sheep.

Prenatal T excess induces maternal hyperinsulinemia, early puberty, and reproductive/metabolic defects in the female similar to those seen in women with polycystic ovary syndrome. This study addressed the organizational/activational role of androgens and insulin in programming pubertal advancement and periovulatory LH surge defects. Treatment groups included the following: 1) control; 2) prenatal T; 3) prenatal T plus prenatal androgen antagonist, flutamide; 4) prenatal T plus prenatal insulin sensitizer, rosiglitazone; 5) prenatal T and postnatal flutamide; 6) prenatal T and postnatal rosiglitazone; and 7) prenatal T and postnatal metformin. Prenatal treatments spanned 30-90 days of gestation and postnatal treatments began at approximately 8 weeks of age and continued throughout. Blood samples were taken twice weekly, beginning at approximately 12 weeks of age to time puberty. Two-hour samples after the synchronization with prostaglandin F2α were taken for 120 hours to characterize LH surge dynamics at 7 and 19 months of age. Prenatal T females entered puberty earlier than controls, and all interventions prevented this advancement. Prenatal T reduced the percentage of animals having LH surge, and females that presented LH surge exhibited delayed timing and dampened amplitude of the LH surge. Prenatal androgen antagonist, but not other interventions, restored LH surges without normalizing the timing of the surge. Normalization of pubertal timing with prenatal/postnatal androgen antagonist and insulin sensitizer interventions suggests that pubertal advancement is programmed by androgenic actions of T involving insulin as a mediary. Restoration of LH surges by cotreatment with androgen antagonist supports androgenic programming at the organizational level.

Chronic Physical Stress Does Not Interact with Epstein-Barr Virus (EBV)-Encoded Dutpase to Alter the Sickness Response.

Most adult humans have been infected with Epstein-Barr virus (EBV), which is thought to contribute to the development of chronic fatigue syndrome. Stress is known to influence the immune system and can exacerbate the sickness response. Although a role for psychological stress in the sickness response, particularly in combination with EBV-encoded deoxyuridine triphosphate nucleotidohydrolase (dUTPase) has been established, and the role of physical stressors in these interactions remains unspecified. In this study, we seek to determine the interaction of chronic physical (swim) stress and EBV-encoded dUTPase injection. We hypothesize that a chronic physical stressor will exacerbate the sickness response following EBV-encoded dUTPase injection. To test this hypothesis mice receive daily injections of EBV-encoded dUTPase or vehicle and are subjected to 15 min of swim stress each day for 14 days or left unmanipulated. On the final evening of injections mice undergo behavioral testing. EBV-encoded dUTPase injection alone produces some sickness behaviors. The physical swimming stress does not alter the sickness response.

Neuroendocrine control of photoperiodic changes in immune function.

Seasonal variation in immune function putatively maximizes survival and reproductive success. Day length (photoperiod) is the most potent signal for time of year. Animals typically organize breeding, growth, and behavior to adapt to spatial and temporal niches. Outside the tropics individuals monitor photoperiod to support adaptations favoring survival and reproductive success. Changes in day length allow anticipation of seasonal changes in temperature and food availability that are critical for reproductive success. Immune function is typically bolstered during winter, whereas reproduction and growth are favored during summer. We provide an overview of how photoperiod influences neuronal function and melatonin secretion, how melatonin acts directly and indirectly to govern seasonal changes in immune function, and the manner by which other neuroendocrine effectors such as glucocorticoids, prolactin, thyroid, and sex steroid hormones modulate seasonal variations in immune function. Potential future research avenues include commensal gut microbiota and light pollution influences on photoperiodic responses.

Exposure to dim light at night during early development increases adult anxiety-like responses.

Early experiences produce effects that may persist throughout life. Therefore, to understand adult phenotype, it is important to investigate the role of early environmental stimuli in adult behavior and health. Artificial light at night (LAN) is an increasingly common phenomenon throughout the world. However, animals, including humans, evolved under dark night conditions. Many studies have revealed affective, immune, and metabolic alterations provoked by aberrant light exposure and subsequent circadian disruption. Pups are receptive to entraining cues from the mother and then light early during development, raising the possibility that the early life light environment may influence subsequent behavior. Thus, to investigate potential influences of early life exposure to LAN on adult phenotype, we exposed mice to dim (~5 lux; full spectrum white light) or dark (~0 lux) nights pre- and/or postnatally. After weaning at 3 weeks of age, all mice were maintained in dark nights until adulthood (9 weeks of age) when behavior was assessed. Mice exposed to dim light in early life increased anxiety-like behavior and fearful responses on the elevated plus maze and passive avoidance tests. These mice also displayed reduced growth rates, which ultimately normalized during adolescence. mRNA expression of brain derived neurotrophic factor (BDNF), a neurotrophin previously linked to early life environment and adult phenotype, was not altered in the prefrontal cortex or hippocampus by early life LAN exposure. Serum corticosterone concentrations were similar between groups at weaning, suggesting that early life LAN does not elicit a long-term physiologic stress response. Dim light exposure did not influence behavior on the open field, novel object, sucrose anhedonia, or forced swim tests. Our data highlight the potential deleterious consequences of low levels of light during early life to development and subsequent behavior. Whether these changes are due to altered maternal behavior or persistent circadian abnormalities incurred by LAN remains to be determined.

Developmental programming: impact of prenatal exposure to bisphenol-A and methoxychlor on steroid feedbacks in sheep.

Bisphenol-A (BPA), a polymer used in plastics manufacturing, and methoxychlor (MXC), a pesticide, are endocrine disrupting compounds with estrogenic and anti-androgenic properties. Prenatal BPA or MXC treatment induces reproductive defects in sheep with BPA causing prepubertal luteinizing hormone (LH) hypersecretion and dampening of periovulatory LH surges and MXC lengthening follicular phase and delaying the LH surge. In this study, we addressed the underlying neuroendocrine defects by testing the following hypotheses: 1) prenatal BPA, but not MXC reduces sensitivity to estradiol and progesterone negative feedback, 2) prenatal BPA, but not MXC increases pituitary responsiveness to gonadotropin releasing hormone (GnRH), and 3) prenatal BPA dampens LH surge response to estradiol positive feedback challenge while prenatal MXC delays the timing of the LH surge. Pregnant sheep were treated with either 1) 5mg/kg/day BPA (produces approximately twice the level found in human circulation, n=8), 2) 5mg/kg/day MXC (the lowest observed effect level stated in the EPA National Toxicology Program's Report; n=6), or 3) vehicle (cotton seed oil: C: n=6) from days 30 to 90 of gestation. Female offspring of these ewes were ovariectomized at 21months of age and tested for progesterone negative, estradiol negative, estradiol positive feedback sensitivities and pituitary responsiveness to GnRH. Results revealed that sensitivity to all 3 feedbacks as well as pituitary responsiveness to GnRH were not altered by either of the prenatal treatments. These findings suggest that the postpubertal reproductive defects seen in these animals may have stemmed from ovarian defects and the steroidal signals emanating from them.

Developmental programming: prenatal and postnatal contribution of androgens and insulin in the reprogramming of estradiol positive feedback disruptions in prenatal testosterone-treated sheep.

Prenatal testosterone (T) excess compromises the estradiol (E(2)) positive feedback. This study tested the hypothesis that antagonizing androgen action or improving insulin sensitivity prenatally would prevent positive feedback disruptions from developing, whereas postnatal intervention with androgen antagonist or insulin sensitizer would ameliorate the severity of disruptions in prenatal T-treated females. The E(2) positive feedback response was tested at 16 wk of age in the following groups of animals: 1) control, 2) prenatal T, 3) prenatal T plus the androgen antagonist, flutamide, 4) prenatal T plus insulin sensitizer, rosiglitazone, 5) prenatal T and postnatal androgen antagonist, and 6) prenatal T and postnatal insulin sensitizer (n = 7-21 animals/group). Prenatal T treatment involved the administration of T propionate (100 mg, im) twice weekly from d 30 to 90 of gestation. Prenatal interventions involved daily sc administration of androgen antagonist (15 mg/kg) or oral administration of insulin sensitizer (8 mg) for the same duration. Postnatal treatments began at 8 wk of age and involved daily oral administration of androgen antagonist (15 mg/kg) or insulin sensitizer (0.11 mg/kg). None of the prenatal/postnatal interventions increased number of animals responding or prevented the time delay in LH surge response to the E(2) positive feedback challenge. In contrast, the postnatal treatment with androgen antagonist or insulin sensitizer increased total LH released in response to E(2) positive feedback challenge, compared with the T animals. Overall, these interventional studies indicate that timing and magnitude of the LH surge are programmed by different neuroendocrine mechanisms with postnatal androgens and insulin determining the size and prenatal estrogen likely the timing of the LH surge.

Combining non-invasive methods for the rapid assessment of mammalian richness in a transect-quadrat survey scheme - Case Study of the Horsh Ehden Nature Reserve, North Lebanon.

Lebanon lacks updated information related to the status of mammalian species, their richness and distribution. This study aimed at developing a rapid assessment methodology combining three non-invasive techniques based on the transect-quadrat survey scheme to measure mammalian richness at the Horsh Ehden Nature Reserve. The achieved results showed that the combination of the three techniques, droppings, footprints and photo-trapping for the rapid assessment of mammalian richness supported by Geographical Information System applications is highly effective. Twenty visits covering twenty quadrats out of 49 over a period of nine months allowed the detection of 12 of the 14 targeted species with droppings providing the majority of evidence while footprints and photo-trapping being of equal efficiency. The method can be easily replicated in any region to rapidly assess mammalian richness and the area of activity of the detected species and therefore directing conservation and management activities towards species of interest.

Interaction between lactation, photoperiodism and male effect in German Merino ewes.

A study with 93 German Merino ewes was performed from January until the end of March to clarify the relative importance of lactation, photoperiodism and ram effect on cyclic activity and lambing data. Ovarian activity was registered by progesterone concentrations in blood plasma three times weekly. Half of the ewes were kept under supplemental light (20 h/day) for the last 6 weeks of lactation and additionally 3 weeks post-weaning, the other half were kept under natural photoperiod but were weaned simultaneously. Thereafter, light was reduced to natural photoperiod and rams were introduced to half of the ewes, of both light reduced and photoperiod group. Ewes entered cyclicity during lactation gradually, but at weaning 56% of photoperiod ewes and 53% of supplemental light ewes were still acyclic. After weaning, resumption of cyclic activity before ram introduction was more pronounced (P<0.05) in the photoperiod group (75% cyclic) than in the supplemental light group (51% cyclic). Ram introduction led to cyclicity in all ewes. Light reduction without ram slightly increased cyclicity but 57% were still acyclic. In the photoperiod group without ram no ewe entered cyclicity and two ewes even ceased cycling again. Data show that German Merinos still have a remarkable lactational anoestrus but are extremely sensitive to ram. Light reduction has no direct effect on cyclicity but is likely to contribute to the elevated ovulation rate so that a combination with the ram effect led to a higher lambing rate (1.94) compared to photoperiod and ram (1.55).