Transcriptional effect of serotonin in the ganglia of Lymnaea stagnalis

The serotonin system (5HT) is highly conserved in both vertebrates and invertebrates, and numerous evidence supports a biological link between 5HT and numerous animal function. In the present paper we evaluated the transcriptional effects of a serotonergic stimulation on selected targets involved in 5HT signalling and neurotransmission in the central nervous system of the great pond snail Lymnaea stagnalis. Adult snails were treated acutely (6 h) or chronically (48 h) with either 5-hydroxytrypthophan (5-HTP 1mM), the immediate precursor of serotonin, fluoxetine (FLX 1μM), a selective serotonin reuptake inhibitor, or a combination of two. The central ring ganglia were dissected and used for q-PCR gene expression analysis. Transcription was strongly induced following a chronic, but not an acute, exposure to 5-HTP in the ganglia of Lymnaea. In particular, LymCREB1 and LymP2X mRNA levels were decreased following a 6 h exposure and increased in snails receiving 5-hydroxytryptophan for 48 h. Interestingly, this effect was reduced when snails were exposed chronically to both 5-HTP and FLX, suggesting a role for SERT in mediating the effect of 5-hydroxytryptophan. These data suggest that L. stagnalis is suited to unravel the complexity of the serotonin signaling pathway

Behavioral and Transcriptional Effects of Short or Prolonged Fasting on the Memory Performances of Lymnaea stagnalis

Introduction: The Garcia effect, a solid learning paradigm, was used to investigate the molecular and behavioral effects induced by different lengths of fasting on the cognitive functions in the pond snail Lymnaea stagnalis, a valid model systemMethods: Three experimental groups were used: Moderately hungry snails, food-deprived for 1 day (D1 snails), severely hungry snails (D5 snails), fasting for 5 days, and satiated snails with ad libitum access to food (AL snails). In the Garcia effect, a single pairing of an appetitive stimulus with a heat stressor results in a learned taste-specific negative hedonic shift. D5 snails were injected with bovine insulin and D1 snails with the insulin receptor antibody (Ab). As a control group, AL snails were injected with saline. Gene expression analyses were performed by Real-time PCR in snails' central nervous system (CNS).Results: AL snails are 'average learners', D1 snails are the best performers, whereas the D5 ones do not show the Garcia effect. Severely fasting snails injected with insulin 3h before the training procedure, show the Garcia effect, whereas injecting 1-day fasting snails with insulin receptor Ab blocks their ability to express memory. The differences in memory performances are associated with changes in the expression levels of selected targets involved in neuronal plasticity, energy homeostasis, and stress response.Discussion: Our results suggest that short-term fasting creates an optimal internal state in L. stagnalis' CNS, allowing a spike in insulin release and an upregulation of genes involved in neuroplasticity. Long-term fasting, instead, upregulates genes involved in energy homeostasis and animal survival

Molecular changes associated with escitalopram response in a stress-based model of depression

Converging evidence points at hypothalamus-pituitary-adrenal (HPA) axis hyperactivity and neuroinflammation as important factors involved in the etiopathogenesis of major depressive disorder (MDD) and in therapeutic efficacy of antidepressants. In this study, we examined the molecular effects associated with a response to a week-long treatment with escitalopram in the chronic escape deficit (CED) model, a validated model of depression based on the induction of an escape deficit after exposure of rats to an unavoidable stress. We confirmed our previous result that a treatment with escitalopram (10 mg/kg) was effective after 7 days in reverting the stress-induced escape deficit in approximately 50% of the animals, separating responders from non-responders. Expression of markers of HPA axis functionality as well as several inflammatory mediators were evaluated in the hypothalamus, a key structure integrating signals from the neuro, immune, endocrine systems. In the hypothalamus of responder animals we observed a decrease in the expression of CRH and its receptors and an increase in GR protein in total and nuclear extracts; this effect was accompanied by a significant decrease in circulating corticosterone in the same cohort. Hypothalamic IL-1\uce\ub2 and TNF\uce\ub1 expression were increased in stressed animals, while CXCL2, IL-6, and ADAM17 mRNA levels were decreased in escitalopram treated rats regardless of the treatment response. These data suggest that efficacy of a one week treatment with escitalopram may be partially mediated by a decrease HPA axis activity, while in the hypothalamus the drug-induced effects on the expression of immune modulators did not correlate with the behavioural outcome

Disease-induced neuroinflammation and depression

Progression of major depression, a multifactorial disorder with a neuroinflammatory signature, seems to be associated with the disruption of body allostasis. High rates of comorbidity between depression and specific medical disorders, such as, stroke, chronic pain conditions, diabetes mellitus, and human immunodeficiency virus (HIV) infection, have been extensively reported. In this review, we discuss how these medical disorders may predispose an individual to develop depression by examining the impact of these disorders on some hallmarks of neuroinflammation known to be impaired in depressed patients: altered permeability of the blood brain barrier, immune cells infiltration, activated microglia, increased cytokines production, and the role of inflammasomes. In all four pathologies, blood brain barrier integrity was altered, allowing the infiltration of peripheral factors, known to activate resident microglia. Evidence indicated morphological changes in the glial population, increased levels of circulating pro-inflammatory cytokines or increased production of these mediators within the brain, all fundamental in neuroinflammation, for the four medical disorders considered. Moreover, activity of the kynurenine pathway appeared to be enhanced. With respect to the inflammasome NLRP3, a new target whose role in neuroinflammation is emerging as being important, accumulating data suggest its involvement in the pathogenesis of brain injury following stroke, chronic pain conditions, diabetes mellitus or in HIV associated immune impairment. Finally, data gathered over the last 10 years, indicate and confirm that depression, stroke, chronic pain, diabetes, and HIV infection share a combination of underlying molecular, cellular and network mechanisms leading to a general increase in the neuroinflammatory burden for the individual

The Development of a Screening Tool for Childcare Professionals to Detect and Refer Infant and Toddler Maltreatment and Trauma: A Tale of Four Countries

Abstract: Child maltreatment is considered a pressing social question, compromising the present and future mental and physical health of one in four children in Europe. While children younger than three years of age are especially vulnerable, few screening instruments are available for the detection of risk in this age group. The purpose of this research was to develop a screening tool for childcare professionals working in public and private daycare settings to support them in the early identification and referral of infants and toddlers exposed to emotional and physical abuse and neglect by primary caregivers, to be used in different settings across four European countries: Belgium, Italy, Latvia, and Hungary. Method: A stratified process was used to create the screening tool: We started by using Living lab methodology to co-create the screening tool with its final users, which was followed by testing the tool with a total of 120 childcare professionals from the four participating countries. Results: During the Living Lab phase, a screening tool with three layers was developed. The initial layer includes five “red flags” that signal particular concern and require immediate action. The second layer is a quick screener with twelve items focused on four areas: neglect of basic needs, delays in development, unusual behaviors, and interaction with caregivers. The third layer is an in-depth questionnaire that aids in formalizing a thorough observation of twenty-five items within the same four areas as the quick screener. After a one-day training session, 120 childcare professionals caring for children aged 0–3 from four countries assessed the screening tool and their overall training experience. Childcare professionals reported great satisfaction with the three-layered structure, which made the tool versatile, and agreed on its content, which was considered helpful in the daycare setting for the regular evaluation of the behavior of children and their primary caregivers, thus improving the early observation of change from the normal behavior of the infant or toddler. Conclusion: The three-layered screening tool was reported as feasible, practical, and with great content validity by childcare professionals working in four European countries

6-Methoxy-2-Benzoxazolinone and photoperiod: prenatal and postnatal influences on reproductive development in prairie voles (Microtus ochrogaster).

The effects of photoperiod and 6-methoxy-2-benzoxazolinone (6-MBOA) on the gonadal development of prairie voles was studied when this plant compound was available to pups in utero, during nursing, or postweaning. In part 1, pregnant voles exposed to long or short day lengths were fed food in which 6-MBOA was present (50 μg of 6-MBOA/g food) or absent prior to the birth of their young; all of these dams received a diet that lacked 6-MBOA after the pups were born. Other females were fed food without 6-MBOA throughout pregnancy, but received one of the diets during lactation. Short days inhibited reproductive development in male offspring, affected pup survival, and enhanced pelage development. Postnatal exposure to 6-MBOA did not affect the reproductive function of males. In contrast, prenatal exposure to 6-MBOA accelerated reproductive development among short-day males. Prenatal exposure to 6-MBOA did not affect survival of short-day males, but reduced survival of long-day pups. Prenatal 6-MBOA treatment affected the sex ratio of pups that survived until weaning (13:21; male:female) and of pups that died prior to weaning (26:5). In contrast, photoperiod did not affect reproductive function in female voles. Although reproductive function appears to be uncoupled from photoperiodic regulation, female prairie voles processed photoperiodic information; pelage development was enhanced among short-day females. In part II, breeding pairs were exposed to long or short days and fed food devoid of 6-MBOA prior to the birth of the young. On the day of birth, and for the following 3 weeks, either 6-MBOA-free or 6-MBOA-enriehed food was provided. After weaning, pups received their parents type of food until autopsy 3 or 7 weeks later. Again, photoperiod affected male, but not female, reproductive function. No consistent pattern of results from postnatal exposure to 6-MBOA was discerned for either males or females. Acceleration of reproductive function among short-day males because of prenatal influences may, be involved in the rapid rate of sexual maturation observed prior to peak population densities of microtine rodents. Because males of this species induce females into estrus, male prairie voles may depend on environmental cues to regulate reproductive function, while females may respond only to males. The physiological and ecological implications of these data are discusse

Effective pain reduction of non-pharmacological interventions for procedural pain during repetitive immunizations of children born pre-term

Psychological distress is common during pregnancy but the field of perinatal-psychology and related research as well as health services have focused their attention essentially on the post-partunl period. New data suggest that stress and psychopathology during pregnancy may be associated with significant risks for the mother and the baby and may lead to detrimental effects for both. Maternal psychopathology is related to reduced quality of life, higher rate of risk behaviors, postpartunl psychopathology, and to a decline in the quality of dyadic relationship. Antenatal stress and psychological disease and their underlying neuroendocrine changes are associated with poor pregnancy and birth outcomes. Maternal stress and psychopathology together with fetal vulnerability and other ~ediating factors, such as pharmachological treatment, may determme long-term consequences by altering developmental processes, affecting the structural development of certain brain areas circuits, and systems, as well as brain functioning. ' ~fter h~ving studied a sample obtained from the general population, thIS study focused on a selected and high risk sample of women diagnosed with psychopathology and recruited from a center specialized in Women's lifecycle mood disorders (psicheDonna Center-Milan). General aim: The general aim of this study was threefold: - to study different manifestations ofpsychological illness as well as to detect risk and protective factors in this selected sample of pregnant and postpartunl women - to analyze the characteristics of these women in order to understand the mechanisms underlying the interaction between protective and risk factors which may predict the course of psychopathological manifestations across pregnancy and beyond. - to analyze, in follow-up, the choice and efficacy of pharmacological and-or psychotherapy treatment of these women. Methods: A sample of91 pregnant and postpartunl women was enrolled from the Center. Women were subjected to a test battery in order to evaluate: - ~ndex ofpregnancy specific-related anxiety (PRAQ-R, Huizink) - mdex of State Anxiety and of Trait Anxiety, as defined by Spielberger (STAI-Y, Spielberger) - ananmestic data and risk and protection factors (scale constructed ad hoc and PDPI, Tatano Beck) - index of depressionlpostpartunl depression (EPDS, Cox). Conclusions and Considerations: Differences were found between ~is sample and the one from the general population, not only m test scores but overall in the role played by risk and protection factors. The presence of pregnancy-related specific anxiety (detected by PRAQ-R) was found more in the general population: a working hypothesis as to the underlying cause was developed. Women recruited from the Center are characterized by a peculiar configuration of risk and protection factors, where a previous history of psychopathology seems to play a prominent role in defining the sample. This type of evidence indicates a possible new key point with respect to intervention and prevention, that is, the previous history of psychopathology. Consequently we need to consider the necessity of different approaches in the use of screening tools, and in the development of preventive measures and healing programs, and tailor all activities and interventions to the patient in order to be effective. This has lead to new evidence-based perspectives for intervention

BetaCasomorphin causes hypoalgesia in 10-day-old-rats:evidence for central mediation

Two experiments determined behavioral effectiveness of \u3b2-casomorphins(\u3b2-CM) in 10-d-old rats by evaluating changes in heat escape latency from a 48\ub0C stimulus applied to a forepaw. In one study rats were injected systemically with \u3b2-CM4, -5, or -7 at a dose range of 0.1-2.5 mg/kg. Only\u3b2-CM5 was effective, and the dose-response relationship was graded. The second study evaluated the locus of action of \u3b2-CM5 through two experimental manipulations: first, by injecting it (0.25 \u3bcg) into the lateral ventricles and by attempting to block its effects with systemic injections of naloxone. Second, rats received intracerebroventricular injections of naloxone (0.25 \u3bcg) and systemic injections of \u3b2-CM.\u3b2-CM was effective centrally, suggesting central detection of the drug. Naloxone injected into the lateral ventricles blocked the effects of systemic administration of \u3b2-CM, implying that circulating \u3b2-CM or their precursors cause behavioral change through central mechanisms

Photoperiodic effects on tumor development and immune function.

Seasonal changes in adaptations associated with winter coping strategies have been frequently studied. Central among the suite of energy-saving, winter-coping strategies is the suspension of reproductive activities. The inhibition of reproduction by nontropical rodents is mediated by daylength changes. Although balanced annual energy budgets are critical, survival and subsequent reproductive success also require avoiding predators, illness, and early death. Because the stressors of winter could lead to suppressed immune function, we hypothesized that animals should have evolved survival strategies involving immunoenhancement. Short daylengths provide a predictive cue to individuals that could be used to enhance immune function in advance of stress-induced immunosuppression. In Experiment 1, adult female deer mice (Peromyscus maniculatus) were housed in either long (LD 16:8) or short (LD 8:16) days for 8 weeks, then injected with the chemical carcinogen 9,10-dimethyl-1,2-benzanthracene (DMBA) dissolved in dimethyl sulfoxide (DMSO) or with the DMSO vehicle alone. Animals were evaluated weekly for 8 weeks after injection. None of the animals treated with DMSO developed tumors in any of the experiments. Nearly 90% of the long-day deer mice injected with DMBA developed squamous cell carcinoma. None of the short-day deer mice injected with DMBA developed tumors. Small lesions developed at the site of injection; short-day females had less severe lesions and healed faster than long-day females. Immunoglobulin G (IgG) response to i.p. injection of sheep red blood cells (SRBC) did not differ photoperiodic conditions. The role of estrogens in the photoperiodic responses was evaluated in Experiment 2: Ovariectomized or sham-ovariectomized deer mice received estradiol benzoate replacement therapy or a control procedure in long daylengths for 8 weeks prior to injection of DMBA or DMSO, then were monitored for 8 additional weeks. Females treated with DMBA developed tumors at the same rate, regardless of estrogen manipulation. Estrogen did not affect healing rates. In Experiment 3, female deer mice were injected with a slurry of microspheres that either contained bromocriptine or were empty. Suppression of prolactin with bromocriptine resulted in a decrease of tumor incidence from 55.6% to 24% in long-day females 8 weeks after injection with DMBA. Healing rates were not affected by prolactin manipulations. Silastic capsules that were filled with either melatonin or cholesterol were implanted into long-day female deer mice in Experiment 4; 8 weeks later, females received an injection of either DMBA or DMSO, then were monitored for 8 weeks.

Day length affects immune cell numbers in deer mice: The influence of age, sex, and in utero photoperiodic history

The extent to which day length affects immune function was examined in the present study. Three goals were pursued: 1) to confirm and extend the observation that the immune systems of adult deer mice (Peromyscus maniculatus) are responsive to changes in photoperiod, 2) to examine the development of the photoperiod-associated changes in immune function, and 3) to discover whether photoperiodic information transmitted to the young during gestation influences immune function. In experiment 1, adult mice housed in short days had higher white blood cell and lymphocyte numbers than their long-day cohorts. Red blood cell and differential cell counts did not differ between long- and short-day animals. No sex differences were observed in the pattern of immune responses to photoperiod. The effect of photoperiod on immune cells in prepubertal animals was examined in experiment 2; a similar pattern of results was obtained as that for experiment 1, suggesting that the photoperiodic effect on the immune system is not mediated by sex steroid hormones. Prenatal and postnatal photoperiodic effects on immune cells were examined in experiment 3; pups gestated in one day length were cross-fostered to mothers in the same day length conditions or to mothers maintained in the alternative day length. The results of experiment 3 suggested that photoperiodic information transmitted from the mother to the young in utero subsequently affected immune systems of the pups. Animals gestated in short day lengths displayed higher immune status throughout life than mice gestated in long days. These results are discussed from an adaptive functional perspective