Stress-mediated hyperactivity and anhedonia resistant to diazepam and fluoxetine in Drosophila

Distresses may induce behavioral phenotypes constituting heuristic models for psychopharmacology studies. In several species, including Drosophila, antidepressants counteract stress-induced phenotypes allowing the use of these models to test new psychoactive drugs. Here, we developed a novel and time-efficient protocol to provoke stress-induced phenotypes in Drosophila for the study of psychopharmacological agents. In the first experiment, flies (n = 12/groups) were exposed to a random-sequence of different types of stresses during nearly 24 h (including social isolation, fasting, heat, and electric shock), a protocol named short-term variable stress (SVS). Second, flies were exposed to a single stressful stimulus (social isolation, fasting, heat shock or electric shock, n = 12/groups). Next, flies submitted to SVS protocol were treated with vehicle, diazepam or fluoxetine (n = 12/groups). At the end of the stress protocols, behavioral phenotypes were evaluated in the open field (OF) and sucrose preference tests. In comparison to the unstressed group, flies exposed to SVS exhibited hyperactivity, as well as shorter times exploring the boundaries of the OF. In contrast to fasting stress, SVS reduced sucrose preference in flies. By analyzing the effects of individual stimuli on fly behavior, fasting and electric shock appear to be the predominant influences on the SVS-induced behaviors. Although fluoxetine or diazepam reduced the initial locomotor activity of flies, no treatment prevented the sequelae of SVS. Altogether, this study provides a time-efficient model system for the study of stress-mediated hyperactivity and anhedonia-like state resistant to fluoxetine and diazepam. The applications of SVS in Drosophila to preclinical psychopharmacology require further studies.LAY SUMMARY Exposition to unpredictable stress plays a significant role in psychiatric disorder’s onset. Behavioral traits of these disorders can be partially modeled in rodents aimed at developing psychopharmacological therapies. However, studies in rodents were questioned by ethical issues. Focused on 3Rs principles, we developed a preclinical model for stress and psychopharmacology research in Drosophila. Variable stress induced behavioral alterations, including hyperlocomotion and reduced preference for sucrose in flies. However, behavioral alterations were resistant to fluoxetine and diazepam.Fil: Ramos Hryb, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Ramirez, Mauro Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Lino de Oliveira, Cilene. Universidade Federal de Santa Catarina; BrasilFil: Pagani, Mario Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentin

Partial Reversal of Striatal Damage by Palmitoylethanolamide Administration Following Perinatal Asphyxia

Perinatal asphyxia (PA) is a clinical condition brought by a birth temporary oxygen deprivation associated with long-term damage in the corpus striatum, one of the most compromised brain areas. Palmitoylethanolamide (PEA) is a neuromodulator well known for its protective effects in brain injury models, including PA, albeit not deeply studied regarding its particular effects in the corpus striatum following PA. Using Bjelke et al. (1991) PA model, full-term pregnant rats were decapitated, and uterus horns were placed in a water bath at 37°C for 19 min. One hour later, the pups were injected with PEA 10 mg/kg s.c., and placed with surrogate mothers. After 30 days, the animals were perfused, and coronal striatal sections were collected to analyze protein-level expression by Western blot and the reactive area by immunohistochemistry for neuron markers: phosphorylated neurofilament-heavy/medium-chain (pNF-H/M) and microtubule-associated protein-2 (MAP-2), and the astrocyte marker, glial fibrillary acidic protein (GFAP). Results indicated that PA produced neuronal damage and morphological changes. Asphyctic rats showed a decrease in pNF-H/M and MAP-2 reactive areas, GFAP+ cells number, and MAP-2 as well as pNF-H/M protein expression in the striatum. Treatment with PEA largely restored the number of GFAP+ cells. Most important, it ameliorated the decrease in pNF-H/M and MAP-2 reactive areas in asphyctic rats. Noticeably, PEA treatment reversed the decrease in MAP-2 protein expression and largely prevented PA-induced decrease in pNF-H/M protein expression. PA did not affect the GFAP protein level. Treatment with PEA attenuated striatal damage induced by PA, suggesting its therapeutic potential for the prevention of neurodevelopmental disorders.Fil: Udovin, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Kobiec, Tamara. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Facultad de Psicología y Psicopedagogía. Centro de Investigaciones en Psicología y Psicopedagogía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Herrera, María Inés. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Facultad de Psicología y Psicopedagogía. Centro de Investigaciones en Psicología y Psicopedagogía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Toro Urrego, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Kusnier, Carlos Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Kolliker Frers, Rodolfo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Ramos Hryb, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Luaces, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Otero-losada, Matilde Estela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Capani, Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina. Universidad Argentina "John F. Kennedy"; Argentin

Efecto neuroprotector de FK506 ante el estrés oxidativo

Las inmunofilinas son proteínas que unen drogas inmunosupresoras. Las que unen ciclosporina A (CsA) son las ciclofilinas (ej: CyPA), y las que reconocen al macrólido FK506 son las FKBPs (FK506-Binding Proteins), una subfamilia a la que pertenecen FKBP51 (51-kDa) y FKBP52 (52-kDa).Fil: Daneri Becerra, Cristina del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Rosbaco, Maria Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Galigniana, Mario Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Ramos Hryb, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaIX Jornadas de Jóvenes InvestigadoresBuenos AiresArgentinaUniversidad de Buenos Aires. Facultad de Ciencias Veterinaria

Neuroprotective effect of FK506 against oxidative stress

Immunophilins FKBP51 and FKBP52 bind the macrolide FK506. Previously, we reported that FK506 favours neurodifferentiation and the neuroregeneration. Here, we analyzed whether FK506 also shows neuroprotective action to oxidative stress. A rapid neuritogenesis was observed in undifferentiated N2a cells treated with 1 µM FK506 in the absence of trophic factors, including serum. Then, 250 µm slices from prefrontal cortexes from Balb-C mice (60 d) were prestabilized for 72 h and incubated for 4 h with 200 µM H2O2.Fil: Rosbaco, Maria Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Daneri Becerra, Cristina del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Galigniana, Mario Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Ramos Hryb, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaXXXIV Anual Meeting SAN 2019Villa Carlos PazArgentinaSociedad Argentina de Investigacion en Neurociencia

Neurorestorative and protective effects of palmitoylethanolamide in perinatal asphyxia: An analysis of the rat striatum

Perinatal asphyxia (PA) caused by low O2 availability during birth is associated with brain damage, been the striatum one of the most affected areas. Palmitoylethanolamide (PEA) is a neuroprotective amide in brain injury models, including PA. However, its effects against PA requires deeper study in the striatum. Using Bjelke? model, full-term pregnant rats were rapidly decapitated, and uterus horns placed in a water bath (37°C,19 min). One hour after, rats were treated with PEA (10 mg/kg,s.c.) and later given to surrogate mothers. 30d animals were perfused and coronal striatum sections as well as subcellular fractions were collected for immunohistochemistry and blotting (n=3). Phosphorylated high/medium molecular weight neurofilaments (pNF-H/M), Microtubule-Associated Protein 2 (MAP2), and Glial fibrillary acidic protein (GFAP) were analysed. All procedures were previously approved by our Institution. 2-way ANOVA followed by Tukey revealed a reduction in pNF-H/M and MAP2 reactive areas (45% and 70%), GFAP+ cells (20%) and reduced MAP2 and pNF-H/M immunocontents in PA-rat striatum. PEA totally restored GFAP+ cells and MAP2 immunoreaction and partially prevented the decreased pNF-H/M reactive area (78%) induced by PA. PEA also reversed the MAP2 reduction and partially prevented the decrease of pNF-H/M immunocontent induced by PA. No GFAP level alteration was detected. PEA treatment attenuated the striatum damage induced by PA, demonstrating its therapeutic potential against PA.Fil: Udovin, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional - Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas "Prof. Dr. Alberto C. Taquini". Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional; ArgentinaFil: Kobiec, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Pontificia Universidad Católica de Argentina Santa María de los Buenos Aires. Facutlad de Piscología y Psicopedagogía. Centro de Investigaciones en Psicología y Pedagogía; ArgentinaFil: Herrera, María Inés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Pontificia Universidad Católica de Argentina Santa María de los Buenos Aires. Facutlad de Piscología y Psicopedagogía. Centro de Investigaciones en Psicología y Pedagogía; ArgentinaFil: Ramos Hryb, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional - Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas "Prof. Dr. Alberto C. Taquini". Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Toro, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional - Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas "Prof. Dr. Alberto C. Taquini". Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional; ArgentinaFil: Kusnier, Carlos Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional - Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas "Prof. Dr. Alberto C. Taquini". Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional; ArgentinaFil: Capani, Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional - Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas "Prof. Dr. Alberto C. Taquini". Instituto Alberto C. Taquini de Investigaciones en Medicina Traslacional; ArgentinaXXXIII Congreso de la Sociedad Argentina de Investigación en NeurocienciasCórdobaArgentinaSociedad Argentina de Investigación en Neurociencia

Regulation of FKBP51 and FKBP52 functions by post-translational modifications

FKBP51 and FKBP52 are two iconic members of the family of peptidyl-prolyl-(cis/trans)-isomerases (EC: 5.2.1.8), which comprises proteins that catalyze the cis/trans isomerization of peptidyl-prolyl peptide bonds in unfolded and partially folded polypeptide chains and native state proteins. Originally, both proteins have been studied as molecular chaperones belonging to the steroid receptor heterocomplex, where they were first discovered. In addition to their expected role in receptor folding and chaperoning, FKBP51 and FKBP52 are also involved in many biological processes, such as signal transduction, transcriptional regulation, protein transport, cancer development, and cell differentiation, just to mention a few examples. Recent studies have revealed that both proteins are subject of post-translational modifications such as phosphorylation, SUMOlyation, and acetylation. In this work, we summarize recent advances in the study of these immunophilins portraying them as scaffolding proteins capable to organize protein heterocomplexes, describing some of their antagonistic properties in the physiology of the cell, and the putative regulation of their properties by those post-translational modifications.Fil: Daneri Becerra, Cristina del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Zgajnar, Nadia Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Lotufo, Cecilia Maricel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Ramos Hryb, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Piwien Pilipuk, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Galigniana, Mario Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentin

Master dissertation: Martins,T

The forced swimming test is a test applied to rodents, commonly used to predict the effects of potentially antidepressant substances. The evaluation of the effect of antidepressants occurs mainly by measuring the duration of the immobility behavior expressed by the rodent, which increases with prolonged exposure to the test, and treatment with antidepressants decrease this passive behavior. Over the years, the test has undergone refinements, accompanied by the insertion of cofactors, from the variation in the population, intervention, and even in the experimental protocols. In addition, naturally, the quality of exploratory studies has been increased, and guides for planning and reporting experiments have their influence on this change. Even with these sources of heterogeneity, the published results of the forced swim test outcomes are promising, raising the question of a possible publication bias, where only positive results are published, and negative ones are suppressed from the literature. Thereby, some questions were raised: "How effective are antidepressants on reducing rodent immobility time in the forced swim test?", "How do the characteristics and quality of studies affect the effect size of antidepressants?" and “Is there a publication bias in the area?”. In the present study, we expected to answer these questions by conducting a systematic review and network meta-analysis

Drosophila as an alternative preclinical model for stress and psychopharmacological research

Preclinical studies of depression and antidepressants currently involve the use of laboratory rodent as animal models. These models mimic behavioral characteristics of the human disorder, such as helplessness, despair, and anhedonia in laboratory animals. Uncontrollable stress may also alter the preference of animals for palatable food, which can be used as a predictive test for antidepressant activity. However, considering the heterogeneous symptomatology of depression and the fact its neurobiology has been not completely elucidated, these animal models still present low validity. Drosophila react to inescapable stress, which indicates their sufficiently complex mechanisms to study neurobiology of stress and depression. Therefore, by applying the ethical principles of animal experimentation such as 3Rs, our project aims to investigate novel methods for antidepressants research by replacing and reducing the use of vertebrates with fruit flies. We aim to create and describe behavioral tests in Drosophila based on the behavioral despair, helplessness, and anhedonia. In addition, we want to validate these tests by analyzing the response to antidepressants of Drosophila, and the possible related structural and biochemical alterations. The results of this research apply the 3Rs principles by promoting the use of Drosophila in the first phases of preclinical studies and development of antidepressants as well as to contribute to a better understanding of the stress response of Drosophila

A protocol for systematic review and meta-analysis of data from preclinical studies employing the forced swimming test

Depressive disorder is a highly prevalent psychiatric disorder that negatively affects the emotional well-being, and also causes a significant impact on health care costs and workplace productivity. Despite the wide range of antidepressants available, they are only marginally effective in patients. Therefore, there is a criticism related to the abundance of “positive results” contrasting with the partial success of antidepressant treatments in clinical trials or in therapeutics. Besides the lack of representativeness in animal models, many different reasons may contribute to the poor translation from preclinical to clinical studies in antidepressant research. Methodological factors such as poor experimental design and low power analysis as well as confirmation bias and publication bias may account for the low reproducibility and poor representation of animal models of depression. All these factors may contribute to the reduced translation of preclinical studies. In this regard, we aim to estimate the influence of methodological qualities on the outcomes of preclinical studies employing animal models and to determine the effect sizes for primary and secondary outcomes according to species, strains, sex, age, via of administration, natural compound class, treatment schedule and protocols of animal models of depression

Construction and reconstruction of brain circuits: normal and pathological axon guidance

Perception of our environment entirely depends on the close interaction between the central and peripheral nervous system. In order to communicate each other, both systems must develop in parallel and in coordination. During development, axonal projections from the CNS as well as the PNS must extend over large distances to reach their appropriate target cells. To do so, they read and follow a series of axon guidance molecules. Interestingly, while these molecules play critical roles in guiding developing axons, they have also been shown to be critical in other major neurodevelopmental processes, such as the migration of cortical progenitors. Currently, a major hurdle for brain repair after injury or neurodegeneration is the absence of axonal regeneration in the mammalian CNS. By contrasts, PNS axons can regenerate. Many hypotheses have been put forward to explain this paradox but recent studies suggest that hacking neurodevelopmental mechanisms may be the key to promote CNS regeneration. Here we provide a seminar report written by trainees attending the second Flagship school held in Alpbach, Austria in September 2018 organized by the International Society for Neurochemistry (ISN) together with the Journal of Neurochemistry (JCN). This advanced school has brought together leaders in the fields of neurodevelopment and regeneration in order to discuss major keystones and future challenges in these respective fields