Monitoraggio e salvaguardia delle razze asinine autoctone della Sardegna attraverso l'impiego di marcatori molecolari

Sardo and Asinara donkey are classified as endangered and critical rispectively (Risk status classification–FAO). Preservation of these genetic resources is of fundamental importance to protect animal biodiversity. To evaluate the genetic variability of Sardinian donkeys and to estimate their relationships with other breeds, we genotyped 130 individuals from 7 italian breeds using 3 FISSR(Fluorescent Inter Simple Sequence Repeat) markers. We also sequenced 120 individuals for 383bp of the mitochondrial HVSI region. Although FISSR have been never used in donkey, they appear to be effective tools for genotyping analysis.In this study, Bayesian cluster analysis underline that the two sardinian breeds are well-separate from other breeds, and high genetic similarity between them.Very different patterns were found between Asinara and Barockesel donkey, despite their phenotypic similarity. In mitochondrial (HVSI region) analysis, AMOVA and Haplotype analysis reveal that Sardo and Asinara donkey are poorly differentiated within Asinara Island, whereas haplotype differences are evident between Sardinia vs Asinara populations. The median joining network shows both typical donkey CLADE1 (Somalicus) and CLADE2 (Nubian) in Sardinian population, with high prevalence of CLADE1, whereas Asinara population has haplotypes assigned exclusively to CLADE2 in Asinara breed. These results suggest the potential use these genetic for conservation plans

Neurosteroidi: i modulatori endogeni delle emozioni

The discovery that facilitation or inhibition of γ aminobutyric acid (GABA)-mediated neurotransmission results in anxiolytic versus anxiogenic, hypnotic versus somnolitic, and anticonvulsant versus convulsant effects, respectively, provided important early insight into the physiology and pharmacology of central GABAergic transmission. This realization, together with subsequent evidence that high-affinity recognition sites for positive and negative allosteric modulators of GABAA receptors are located on these GABA-gated Cl– channels, led to the concept that GABAA receptors contribute directly not only to the pharmacology but also to the neurobiology and physiopathology of a variety of neurological and psychiatric diseases characterized by changes in emotional state, sleep pattern, or neuronal excitability. These findings have suggested the hypothesis that the brain and peripheral organs in mammals might produce endogenous compounds that selectively modulate central GABAA receptor function. Evidence directly supporting this hypothesis has been provided over the last decade by the discovery that steroid hormones synthesized in the brain or in peripheral organs are among the most selective, potent, and efficacious allosteric modulators of GABAA receptors. Neurosteroids are steroid derivatives that are synthesized de novo from cholesterol in the central nervous system (CNS), some of which modulate GABAA receptor function with potencies and efficacies similar to or greater than those of benzodiazepines and barbiturates. These molecules have thus been suggested to be the endogenous modulators of GABAA receptor–mediated neurotransmission. In fact some of these molecules have the capability to modulate synaptic activity by binding to membrane sites associated with ligand-gated ionotropic receptors including GABAA receptors. Here we summarize some of the most recent evidences obtained by our and other laboratories pertaining the role of two neuroactive steroids allopregnanolone (AP) and tetrahydrodeoxycorticosterone (THDOC) actives in modulating the function and plasticity of GABAA receptors in nature

International Expert Opinions and Recommendations on the Use of Melatonin in the Treatment of Insomnia and Circadian Sleep Disturbances in Adult Neuropsychiatric Disorders

Introduction: Insomnia and circadian rhythm disorders, such as the delayed sleep phase syndrome, are frequent in psychiatric disorders and their evaluation and management in early stages should be a priority. The aim of this paper was to express recommendations on the use of exogenous melatonin, which exhibits both chronobiotic and sleep-promoting actions, for the treatment of these sleep disturbances in psychiatric disorders.Methods: To this aim, we conducted a systematic review according to PRISMA on the use of melatonin for the treatment of insomnia and circadian sleep disorders in neuropsychiatry. We expressed recommendations for the use of melatonin in psychiatric clinical practice for each disorder using the RAND/UCLA appropriateness method.Results: We selected 41 studies, which included mood disorders, schizophrenia, substance use disorders, attention deficit hyperactivity disorders, autism spectrum disorders, neurocognitive disorders, and delirium; no studies were found for both anxiety and eating disorders.Conclusion: The administration of prolonged release melatonin at 2–10 mg, 1–2 h before bedtime, might be used in the treatment of insomnia symptoms or comorbid insomnia in mood disorders, schizophrenia, in adults with autism spectrum disorders, neurocognitive disorders and during sedative-hypnotics discontinuation. Immediate release melatonin at <1 mg might be useful in the treatment of circadian sleep disturbances of neuropsychiatric disorders

Vagus nerve stimulation increases norepinephrine concentration and the gene expression of BDNF and bFGF in the rat brain

Vagus nerve stimulation therapy, effective for treatment-resistant epilepsy, has recently been approved also for treatment-resistant depression; nevertheless, the molecular mechanism(s) underlying its therapeutic action remains unclear. Given that neurotrophic factors and monoamines could play a crucial role in the pathophysiology of depression, we tested whether vagus nerve stimulation increases the expression of brain-derived neurotrophic factor, fibroblast growth factor, and nerve growth factor as well as the concentration of norepinephrine in the rat brain. Rats were implanted with a vagus nerve stimulator device and the effects of acute stimulation were evaluated on the growth factors mRNA levels and norepinephrine concentration by ribonuclease protection assay and microdialysis, respectively. We found that acute vagus nerve stimulation increased the expression of brain-derived neurotrophic factor and fibroblast growth factor in the hippocampus and cerebral cortex, decreased the abundance of nerve growth factor mRNA in the hippocampus, and, similar to the antidepressant drug venlafaxine, increased the norepinephrine concentration in the prefrontal cortex. This study demonstrates that acute vagus nerve stimulation triggers neurochemical and molecular changes in the rat brain involving neurotransmitters and growth factors known to play a crucial role in neuronal trophism. These new findings contribute to the elucidation of the molecular mechanisms underlying the therapeutic actions of vagus nerve stimulation in both treatment-resistant depression and epilep

Increased Voluntary Ethanol Consumption and Changes in Hippocampal Synaptic Plasticity in Isolated C57BL/6J Mice

Social isolation (SI) is a notable model of prolonged mild stress, characterized by multiple neurochemical and behavioral alterations, that appears particularly suitable for studying different aspects of the interplay between stress and ethanol (EtOH) consumption in order to characterize potential molecular mechanisms, including changes in the function of inhibitory GABAergic synapses, underlying such interaction. In C57BL/6J mice, SI is associated with an altered hippocampal concentration of the neuroactive steroids 3α-hydroxy-5α-pregnan-20-one (3α-5α-THP), an increased expression of the α4 and δ subunit of γ-aminobutyric acid type A receptors (GABAARs) in the dentate gyrus (DG), and a parallel enhancement of the stimulatory action of 4,5,6,7-tetrahydroisoxazolo[5,4-c] pyridin-3-ol (THIP) on GABAergic tonic currents recorded in voltage-clamped DG granule cells (DGGCs). In addition, SI in C57BL/6J mice determines an increase in voluntary EtOH consumption and EtOH preference when compared to group-housed (GH) control animals. Furthermore, in hippocampal slices of SI mice we also observed a marked reduction of both cellular excitability and long term potentiation (LTP) in pyramidal neurons of the CA1 hippocampal sub-region, effects that were prevented by the long term treatment of SI mice with the neuroactive steroid precursor progesterone. In this article, we summarize some of our recent findings on the effects of SI in C57BL/6J mice on voluntary EtOH intake, regulation of GABAARs gene expression and function and hippocampal long term synaptic plasticity

Increase in expression of the GABA_A receptor alpha₄ subunit gene induced by withdrawal of, but not by long-term treatment with, benzodiazepine full or partial agonists

The effects of long-term exposure to, and subsequent withdrawal of, diazepam or imidazenil (full and partial agonists of the benzodiazepine receptor, respectively) on the abundance of GABAA receptor subunit mRNAs and peptides were investigated in rat cerebellar granule cells in culture. Exposure of cells to 10 μM diazepam for 5 days significantly reduced the amounts of α4 and γ2 subunit mRNAs, and had no effect on the amount of α1 mRNA. These effects were accompanied by a decrease in the levels of α1 and γ2 protein and by a reduction in the efficacy of diazepam with regard to potentiation of GABA-evoked Cl- current. Similar long-term treatment with 10 M imidazenil significantly reduced the abundance of only the γ2S subunit mRNA and had no effect on GABAA receptor function. Withdrawal of diazepam or imidazenil induced a marked increase in the amount of α4 mRNA; withdrawal of imidazenil also reduced the amounts of α1 and γ2 mRNAs. In addition, withdrawal of diazepam or imidazenil was associated with a reduced ability of diazepam to potentiate GABAA action. These data give new insights into the different molecular events related to GABAA receptor gene expression and function produced by chronic treatment and withdrawal of benzodiazepines with full or partial agonist properties

The metabolic profile of Asinara (albino) and Sardo donkeys (pigmented) (Equus asinus L., 1758) points to unequivocal breed assignment of individuals

This study pointed to explore if variations in circulating levels of metabolites in the blood stream of no. 25 feral donkeys occur in view of the different coat color between specimens of Asinara (albino, no. 8) vs. Sardo (dun-grey, no. 17) breed. All individuals involved in this investigation are living in the nature, at Mediterranean latitudes and roam in the same areas all over the National Park of Capo Caccia, where they feed on spontaneous vegetation sources. The study was conducted during the positive photoperiod of the boreal hemisphere (peak in the month of June, 2019) to maximize the effect of exposure to the natural sun radiation and thus elicit the coping ability of albino (Asinara) in comparison with pigmented donkeys (Sardo). The biochemical profile of all donkeys was used in a Discriminant Analysis (DA) to explore if circulating levels of metabolites could point to metabolic markers for breed assignment of individuals following a canonical discriminant analysis (CANDISC). The biochemical investigation included also the determination of the circulating Vitamin E (alpha tocopherol, α-TOH), as an essential biologically active compound involved in antioxidant mechanisms, and its respective status (circulating α-TOH to total triglycerides and total cholesterol ratio). In the CANDISC, the distance between the two breeds was not significant. However, it pointed to different metabolites (UREA, total protein, total triglycerides, Zn) capable of describing biochemical patterns on each respective breed (Asinara vs. Sardo). The multivariate analysis DA carried out using 22 metabolites correctly assigned individuals to the two breeds in the 100% of cases. In view of such metabolic background, circulating α-TOH found in the bloodstream of Asinara vs. Sardo donkeys under free grazing conditions turned out to reach similar values (2.114 vs. 1.872 µg/ml, respectively, p = 0.676). It is worth noting that significant differences were observed as to circulating lactate dehydrogenase (LDH, p = 0.022) levels, in association with increased creatine phosphokinase (CPK, p = 0.076), both above the upper limit of the physiological range reported in other donkey breeds, and found in the totality of Asinara (albino) donkeys solely, still apparently clinically healthy

Enhanced glutamatergic synaptic plasticity in the hippocampal CA1 field of food-restricted rats: involvement of CB1 receptors

The endogenous endocannabinoid system has a crucial role in regulating appetite and feeding behavior in mammals, as well as working memory and reward mechanisms. In order to elucidate the possible role of cannabinoid type-1 receptors (CB1Rs) in the regulation of hippocampal plasticity in animals exposed to food restriction (FR), we limited the availability of food to a 2-h daily period for 3 weeks in Sprague-Dawley rats. FR rats showed a higher long-term potentiation at hippocampal CA1 excitatory synapses with a parallel increase in glutamate release when compared with animals fed ad libitum. FR rats showed a significant increase in the long-term spatial memory determined by Barnes maze. FR was also associated with a decreased inhibitory effect of the CB1R agonist win55,212-2 on glutamatergic field excitatory postsynaptic potentials, together with a decrease in hippocampal CB1R protein expression. In addition, hippocampal brain-derived neurotrophic factor protein levels and mushroom dendritic spine density were significantly enhanced in FR rats. Altogether, our data suggest that alterations of hippocampal CB1R expression and function in FR rats are associated with dendritic spine remodeling and functional potentiation of CA1 excitatory synapses, and these findings are consistent with increasing evidence supporting the idea that FR may improve cognitive functions.Neuropsychopharmacology advance online publication, 21 October 2015; doi:10.1038/npp.2015.280