Corticosterone dynamically regulates retrotransposable element expression in the rat hippocampus and C6 cells.

The hippocampus is a highly plastic brain region sensitive to environmental stress. It shows dynamic changes in epigenetic marks associated with stress related learning. Previous work has shown that acute stress induces substantial transient changes in histone H3 lysine 9 trimethylation (H3K9me3). Moreover, increased H3K9me3 is enriched in hippocampal gene deserts accumulating within endogenous retroviruses and transposable elements. We have found that in response to acute glucocorticoid treatment, a similar change in global H3K9me3 is observed. However, when localized we found that H3K9me3 is markedly decreased at B2 short interspersed nuclear elements but not within intracisternal-A particle endogenous retroviruses. Further, decreased H3K9me3 valence within B2 elements was associated with increased transcript abundance. These data demonstrate the capacity for acute glucocorticoids to mobilize transposable elements via epigenetic unmasking. Reconciled with previous findings following acute stress, this suggests the capacity for mobile elements to potentially function as novel regulators given their dynamic regulation by stress and glucocorticoids.

Molecular endocrinology of female reproductive behavior.

Epigenetic methodologies address mechanisms of estrogenic effects on hypothalamic and preoptic neurons, as well as mechanisms by which stress can interfere with female reproductive behaviors. Recent results are reviewed.

Neuroepigenetics of stress.

Stress, a common if unpredictable life event, can have pronounced effects on physiology and behavior. Individuals show wide variation in stress susceptibility and resilience, which are only partially explained by variations in coding genes. Developmental programing of the hypothalamic-pituitary-adrenal stress axis provides part of the explanation for this variance. Epigenetic approaches have successfully helped fill the explanatory gaps between the influences of gene and environment on stress responsiveness, and differences in the sequelae of stress across individuals and generations. Stress and the stress axis interacts bi-directionally with epigenetic marks within the brain. It is now clear that exposure to stress, particularly in early life, has both acute and lasting effects on these marks. They in turn influence cognitive function and behavior, as well as the risk for suicide and psychiatric disorders across the lifespan and, in some cases, unto future generations.

Hippocampal gene expression changes underlying stress sensitization and recovery.

Chronic and acute stressors have been linked to changes in hippocampal function and anxiety-like behaviors. Both produce changes in gene expression, but the extent to which these changes endure beyond the end of stress remains poorly understood. As an essential first step to characterize abnormal patterns of gene expression after stress, this study demonstrates how chronic restraint stress (CRS) modulates gene expression in response to a novel stressor in the hippocampus of wild-type mice and the extent to which these changes last beyond the end of CRS. Male C57/bl6 mice were subjected to (1) a forced swim test (FST), (2) corticosterone (Cort) or vehicle injections, (3) CRS for 21 days and then a FST, or (4) allowed to recover 21 days after CRS and subjected to FST. Hippocampal mRNA was extracted and used to generate cDNA libraries for microarray hybridization. Naive acute stressors (FST and vehicle injection) altered similar sets of genes, but Cort treatment produced a profile that was distinct from both FST and vehicle. Exposure to a novel stress after CRS activated substantially more and different genes than naive exposure. Most genes increased by CRS were decreased after recovery but many remained altered and did not return to baseline. Pathway analysis identified significant clusters of differentially expressed genes across conditions, most notably the nuclear factor kappa-light-chain-enhancer of B cells (NF-κB) pathway. Quantitative reverse transcription-PCR (qRT-PCR) validated changes from the microarrays in known stress-induced genes and confirmed alterations in the NF-κB pathway genes, Nfkbia, RelA and Nfkb1. FST increased anxiety-like behavior in both the naive and recovery from CRS conditions, but not in mice 24h subsequent to their CRS exposure. These findings suggest that the effects of naive stress are distinct from Cort elevation, and that a history of stress exposure can permanently alter gene expression patterns in the hippocampus and the behavioral response to a novel stressor. These findings establish a baseline profile of normal recovery and adaptation to stress. Importantly, they will serve as a conceptual basis to facilitate the future study of the cellular and regional basis of gene expression changes that lead to impaired recovery from stress, such as those that occur in mood and anxiety disorders.

Relationships among estrogen receptor, oxytocin and vasopressin gene expression and social interaction in male mice.

The incidence of social disorders such as autism and schizophrenia is significantly higher in males, and the presentation more severe, than in females. This suggests the possible contribution of sex hormones to the development of these psychiatric disorders. There is also evidence that these disorders are highly heritable. To contribute toward our understanding of the mechanisms underlying social behaviors, particularly social interaction, we assessed the relationship of social interaction with gene expression for two neuropeptides, oxytocin (OT) and arginine vasopressin (AVP), using adult male mice. Social interaction was positively correlated with: oxytocin receptor (OTR) and vasopressin receptor (V1aR) mRNA expression in the medial amygdala; and OT and AVP mRNA expression in the paraventricular nucleus of the hypothalamus (PVN). When mice representing extremes of social interaction were compared, all of these mRNAs were more highly expressed in high social interaction mice than in low social interaction mice. OTR and V1aR mRNAs were highly correlated with estrogen receptor α (ERα) mRNA in the medial amygdala, and OT and AVP mRNAs with estrogen receptor ß (ERß) mRNA in the PVN, indicating that OT and AVP systems are tightly regulated by estrogen receptors. A significant difference in the level of ERα mRNA in the medial amygdala between high and low social interaction mice was also observed. These results support the hypothesis that variations of estrogen receptor levels are associated with differences in social interaction through the OT and AVP systems, by upregulating gene expression for those peptides and their receptors.

Structural and functional alterations to rat medial prefrontal cortex following chronic restraint stress and recovery.

Chronic stress has been shown in animal models to result in altered dendritic morphology of pyramidal neurons of the medial prefrontal cortex (mPFC). It has been hypothesized that the stress-induced dendritic retractions and spine loss lead to disrupted connectivity that results in stress-induced functional impairment of mPFC. While these alterations were initially viewed as a neurodegenerative event, it has recently been established that stress induced dendritic alterations are reversible if animals are given time to recover from chronic stress. However, whether spine growth accompanies dendritic extension remains to be demonstrated. It is also not known if recovery-phase dendritic extension allows for re-establishment of functional capacity. The goal of this study, therefore, was to characterize the structural and functional effects of chronic stress and recovery on the infralimbic (IL) region of the rat mPFC. We compared neuronal morphology of IL layer V pyramidal neurons from male Sprague-Dawley rats subjected to 21 days of chronic restraint stress (CRS) to those that experienced CRS followed by a 21 day recovery period. Layer V pyramidal cell functional capacity was assessed by intra-IL long-term potentiation (LTP) both in the absence and presence of SKF38393, a dopamine receptor partial agonist and a known PFC LTP modulator. We found that stress-induced IL apical dendritic retraction and spine loss co-occur with receptor-mediated impairments to catecholaminergic facilitation of synaptic plasticity. We also found that while post-stress recovery did not reverse distal dendritic retraction, it did result in over extension of proximal dendritic arbors and spine growth as well as a full reversal of CRS-induced impairments to catecholaminergic-mediated synaptic plasticity. Our results support the hypothesis that disease-related PFC dysfunction is a consequence of network disruption secondary to altered structural and functional plasticity and that circuitry reestablishment may underlie elements of recovery. Accordingly, we believe that pharmacological treatments targeted at preventing dendritic retraction and spine loss or encouraging circuitry re-establishment and stabilization may be advantageous in the prevention and treatment of mood and anxiety disorders.

Effect of corticosterone on CART peptide levels in rat blood.

We have recently demonstrated that CART peptides display a diurnal rhythm in blood that depends partly on glucocorticoids levels. This study extends previous findings by directly testing the effects of acute administration of corticosterone and metyrapone on CART peptide levels in blood. Acute treatment with corticosterone augmented CART levels, while metyrapone administration prevented the increase in CART in the evening hours. These results further support the hypothesis that glucocorticoids play a role in the regulation of CART levels in blood.

Cocaine- and amphetamine-regulated transcript peptide levels in blood exhibit a diurnal rhythm: regulation by glucocorticoids.

Cocaine- and amphetamine-regulated transcript (CART) peptides are novel neurotransmitters that are implicated in several physiological functions such as control of feeding behavior, drug reward, sensory processing, stress, and development. Although a majority of studies have examined the role of CART in the brain, less is known about its function in the periphery. Therefore, the goals of this study were to examine the levels and species of CART peptides in blood, to determine whether they undergo diurnal rhythms, and to elucidate their sources and regulatory factors. RIA showed that CART peptides are present in the blood of rats and monkeys and that they exhibit a diurnal variation. Western blotting confirmed the pattern of diurnal variation in rats and, additionally, showed that CART immunoreactivity was due to a single predominant fragment with an apparent molecular weight in the range of the active CART 55-102 peptide. Adrenalectomy caused a 70% reduction in CART peptide levels in rat blood, and this was reversed by corticosterone replacement. CART levels paralleled glucocorticoid levels in rat and monkey blood. Control of CART levels by corticosterone suggests the possibility that CART peptides in blood may be influenced by hypothalamic-pituitary-adrenal interactions and that they may play a role in glucocorticoid-related processes such as stress.

CART peptides are modulators of mesolimbic dopamine and psychostimulants.

CART peptide produces behavioral effects when injected into the VTA or nucleus accumbens. In the VTA, the peptide behaves like an endogenous psychostimulant and produces increased locomotor activity and conditioned place preference. Since this is blocked by dopamine receptor blockers, it presumably involves release of dopamine. But in the nucleus accumbens, CART peptide reduces the locomotor-increasing effects of cocaine. This suggests that the peptide is an interesting target for medications development.

Nitrogen, phosphorous, and organic carbon removal in simulated wetland treatment systems.

Effects of vegetation, wastewater drawdown, hydraulic retention time (HRT), and media depth on removal of nitrogen, phosphorous, and organic carbon in microcosms were investigated. Synthetic wastewater was added daily to 28 microcosms, and effluent was sampled every 12 days for 132 days. Effluent was analyzed for ammonium (NH(4)(+)-N), nitrate (NO(3)(-)-N), orthophosphate (PO(4)(3-)-P), and total organic carbon (TOC). Average percent removal of NH(4)(+)-N was significantly greater in microcosms containing plants (67%) than in those without plants (29%). Percent removal of PO(4)(3-)-P was also significantly greater in microcosms with plants (42%) than in microcosms without plants (20%), but no significant difference was found for TOC removal between microcosms with plants (67%) and those without plants (74%). Average removal was significantly lower in microcosms with wastewater drawdown than in those without wastewater fluctuation for both NH(4)(+)-N (51% versus 83%) and PO(4)(3-)-P (14% versus 71%). Percent NH(4)(+)-N removal was significantly greater in microcosms with a 6-day retention time (80%) than in those with a 2-day retention (53%), and PO(4)(3-)-P removal was also significantly greater with a 6-day retention time (55%) than a 2-day retention (29%). No differences were seen in TOC removal due to any of the treatments but HRT, where removal was greater microcosms with a 2-day HRT (76%) than in those with a 6-day HRT (60%). Media depth did not have a significant effect on nutrient removal. Results of this study demonstrate that required design parameters are different depending on the nutrient being removed in systems simulated by these microcosms.

Intra-ventral tegmental area injection of rat cocaine and amphetamine-regulated transcript peptide 55-102 induces locomotor activity and promotes conditioned place preference.

Cocaine- and amphetamine-regulated transcript (CART) is a novel mRNA that has been reported to be increased by acute psychostimulant administration, and that may be involved in the effects of psychostimulants. In this study, we examined the effect of centrally administered CART peptides on locomotor activity and conditioned place preference in the rat. CART peptide fragments were bilaterally injected into the ventral tegmental area. CART 55-102 (0.2-5.0 microg/side), an endogenously occurring peptide, dose dependently increased locomotor activity, whereas CART 1-26 (0.1-2.5 microg/side; not found endogenously) did not. The locomotor effects of CART 55-102 were dose dependently blocked by the dopamine D(2) receptor antagonist haloperidol (0.03-1.0 mg/kg i.p.). Four injections of 1.0 microg/side CART 55-102 induced a significant place preference, suggesting that CART 55-102 is reinforcing. Increases in locomotor activity after each of these CART 55-102 injections were similar and did not show tolerance or sensitization. This treatment regimen of CART 55-102 also did not produce sensitization to locomotor activity after a subsequent challenge with cocaine or amphetamine. When CART 55-102 (0.2-1.0 microg/side) was injected into the substantia nigra, no significant change in motor activity was observed. However, a higher dose of CART 55-102 (5.0 microg/side) induced a delayed increase in motor activity, suggesting a possible diffusion from the substantia nigra into the ventral tegmental area. Our findings suggest that CART 55-102 is behaviorally active and may be involved in the actions of psychostimulants. This is the first demonstration of the psychostimulant-like effects of CART peptides.

CART peptides.

CART peptides are among the newest putative peptide neurotransmitter/cotransmitters. They show no significant homology to any other peptide, and they are thought to have a role in reward and reinforcement, feeding, development, sensory processing, stress and endocrine control.

CART: from gene to function.

CART was identified as a novel mRNA regulated by psychostimulant drugs. CART peptides appear to be neurotransmitters involved in a variety of functions such as feeding. The mouse gene has been characterized and localized to Chromosome 13. The processing of CART peptides is evident in Western blotting studies.

Studies of selected phenyltropanes at monoamine transporters.

3-Phenyltropane analogues of cocaine are useful neurobiologic tools for examining mechanisms of neurotransmitter transporters and psychostimulant drugs. They are also potential substitute medications for psychostimulant abuse. In this study, 18 3-phenyltropane analogues were characterized in uptake and binding studies at dopamine (DAT), norepinephrine (NET) and serotonin (SERT) transporters from the rat, and in binding at DAT in rat, rhesus monkey, and human brain tissue. In rat brain tissue, potency in inhibiting uptake generally correlated with the potency in inhibiting binding at all three transporters suggesting that none of these compounds have antagonist properties. At the DAT, there was a significant correlation of inhibitory potencies between the rat and monkey, the monkey and human, and the rat and human transporters although some compounds showed some species difference. These findings suggest that with regard to the 3-phenyltropane series, there is generally little pharmacologic difference between DATs from the three species examined, although binding data from rat may not be a perfect predictor of uptake inhibition in human.

Pilot study of myoblast transfer in the treatment of Becker muscular dystrophy.

We evaluated myoblast implantation therapy in three subjects with Becker muscular dystrophy who received 60 million myoblasts in one tibialis anterior (TA) muscle 2 months after beginning cyclosporine immunosuppression (5 to 10 mg/kg) that continued for 1 year. Strength of the implanted and control TA muscles was measured before and after treatment using a gauge to record TA contraction force. Our protocol controlled for the effects of cyclosporine and myoblast injections. In this pilot study, myoblast implantation did not improve strength of the implanted TA muscles.

Preservation of the alveolar ridge with hydroxylapatite tooth root substitutes.

A total of 49 patients, who were followed for 3 to 31 months, have had 92 hydroxylapatite root implants placed in fresh extraction sockets without soft tissue closure. Hydroxylapatite root implants maintain approximately twice as much alveolar bone as the contralateral control sites without implants. It is believed that this method is a more effective and efficient procedure to preserve alveolar bone for retention of dentures than previously described procedures.

Organochlorine residues in fish of Lake Texoma, October 1979.

Fillets from 99 fish, representing 11 species and three areas within Lake Texoma, were examined for residues of common organochlorines. Statistical analyses were conducted to determine the relationship of residues in fish fillets to trophic level and to geographical location of the sample. Most species contained PCBs at levels up to 100 ng/g and p,p'-DDE as high as 127 ng/g. p,p'-TDE and o,p'-DDT were not found in carnivores, but were present in herbivores and detritivores in amounts up to 36 ng p,p'-TDE/g and 17 ng o,p'-DDT/g. Heptachlor also was not found in carnivores, but was present as high as 37 ng/g in the other two classes. Chlordane ranged to 24 ng/g and was detected in all trophic levels. Dieldrin and p,p'-DDT were present in detritivores and carnivores up to 144 ng dieldrin/g and 410 ng p,p'-DDT/g. Neither substance was found in herbivores. Mirex, endrin, and heptachlor epoxide were present only at low levels. Statistically significant differences (P = 0.05) were found between trophic levels for seven of the eleven organochlorine compounds. No correlation (P = 0.05) was found between fillet concentrations of any parameter and geographical location of the sample.