Publicaciones por año
Publicaciones del Departamento de Histología y Embriología
Mitochondrial Modulation by Dichloroacetate Reduces Toxicity of Aberrant Glial Cells and Gliosis in the SOD1G93A Rat Model of Amyotrophic Lateral Sclerosis
Neurotherapeutics 2019 16(1):203-215
Laura Martínez-Palma 1 2 , Ernesto Miquel 3 4 , Valentina Lagos-Rodríguez 3 4 , Luis Barbeito 5 , Adriana Cassina 4 6 , Patricia Cassina 7 8
1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. lmartinezpalma07@gmail.com. 2 Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. lmartinezpalma07@gmail.com. 3 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. 4 Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. 5 Institut Pasteur de Montevideo, Mataojo 2020, 11400, Montevideo, Uruguay. 6 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. 7 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. pcassina@fmed.edu.uy. 8 Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Av. Gral Flores 2125, 11800, Montevideo, Uruguay. pcassina@fmed.edu.uy.
DOI: 10.1007/s13311-018-0659-7
PMID: 30159850
Pubmed: https://pubmed.ncbi.nlm.nih.gov/30159850
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S1878-7479(23)01010-3
Abstract:
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by motor neuron (MN) degeneration and gliosis. Neonatal astrocytes obtained from the SOD1G93A rat model of ALS exhibit mitochondrial dysfunction and neurotoxicity that can be reduced by dichloroacetate (DCA), a metabolic modulator that has been used in humans, and shows beneficial effects on disease outcome in SOD1G93A mice. Aberrant glial cells (AbGC) isolated from the spinal cords of adult paralytic SOD1G93A rats exhibit highly proliferative and neurotoxic properties and may contribute to disease progression. Here we analyze the mitochondrial activity of AbGC and whether metabolic modulation would modify their phenotypic profile. Our studies revealed fragmented mitochondria and lower respiratory control ratio in AbGC compared to neonatal SOD1G93A and nontransgenic rat astrocytes. DCA (5 mM) exposure improved AbGC mitochondrial function, reduced their proliferative rate, and importantly, decreased their toxicity to MNs. Furthermore, oral DCA administration (100 mg/kg, 10 days) to symptomatic SOD1G93A rats reduced MN degeneration, gliosis, and the number of GFAP/S100β double-labeled hypertrophic glial cells in the spinal cord. DCA treatment of AbGC reduced extracellular lactate levels indicating that the main recognized DCA action, targeting the pyruvate dehydrogenase kinase/pyruvate dehydrogenase complex, may underlie our findings. Our results show that AbGC metabolic phenotype is related to their toxicity to MNs and indicate that its modulation can reduce glial mediated pathology in the spinal cord. Together with previous findings, these results further support glial metabolic modulation as a valid therapeutic strategy in ALS.
First ovarian response to gonadotrophin stimulation in rats exposed to neonatal androgen excess
J Mol Histol 2018 49(6):631-637
Rebeca Chávez-Genaro 1 , Gabriel Anesetti 2
1 Histology and Embryology Department, School of Medicine, UdelaR, General Flores 2125, CP 11800, Montevideo, Uruguay. rchavez@fmed.edu.uy. 2 Histology and Embryology Department, School of Medicine, UdelaR, General Flores 2125, CP 11800, Montevideo, Uruguay.
DOI: 10.1007/s10735-018-9800-5
PMID: 30302594
Pubmed: https://pubmed.ncbi.nlm.nih.gov/30302594
Texto completo: https://doi.org/10.1007/s10735-018-9800-5
Abstract:
This study analyzes the effects of neonatal androgenization on follicular growth and first ovulation in response to gonadotrophins, using a model of exogenous stimulation or the use of subcutaneous ovary grafts in castrated animals to replace the hypothalamus-pituitary signal. Neonatal rats (days 1-5) were treated with testosterone, dihydrotestosterone or vehicle. At juvenile period, rats were stimulated with PMSG, hCG (alone or combined) or used as ovarian donors to be grafted on castrated adult female rats. Ovulation and ovarian histology were analyzed in both groups. Animals treated with vehicle or dihydrotestosterone stimulated with gonadotrophins (pharmacological or by using an ovary graft) ovulated, showing a normal histological morphology whereas rats exposed to testosterone and injected with the same doses of gonadotrophins did not it. In this group, ovulation was reached using a higher dose of hCG. Ovaries in the testosterone group were characterized by the presence of follicles with atretic appearance and a larger size than those observed in control or dihydrotestosterone groups. A similar appearance was observed in testosterone ovary grafts although luteinization and some corpora lutea were also identified. Our findings suggest that neonatal exposure to aromatizable androgens induces a more drastic signalling on the ovarian tissue that those driven by non-aromatizable androgens in response to gonadotrophins.
Impact of monomeric, oligomeric and fibrillar alpha-synuclein on astrocyte reactivity and toxicity to neurons
Biochem J 2018 475(19):3153-3169
Cecilia Chavarría 1 , Sebastián Rodríguez-Bottero 2 , Celia Quijano 1 , Patricia Cassina 2 , José M Souza 3
1 Departamento de Bioquímica, Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Av. Gral. Flores 2125, Montevideo 11400, Uruguay. 3 Departamento de Bioquímica, Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Montevideo, Uruguay jsouza@fmed.edu.uy.
DOI: 10.1042/BCJ20180297
PMID: 30185433
Pubmed: https://pubmed.ncbi.nlm.nih.gov/30185433
Texto completo: https://portlandpress.com/biochemj/article-lookup/doi/10.1042/BCJ20180297
Abstract:
Synucleinopathies are a group of neurodegenerative disorders characterized by the presence of aggregated and fibrillar forms of alpha-synuclein (α-syn). Here, we analyze the effect of different species of α-syn, including monomeric, oligomeric and fibrillar forms of the protein, on rat astrocytes. Astrocytes treated with these distinct forms of α-syn showed an increase in long and thin processes and glial fibrillary acidic protein expression, indicating cell activation, high levels of intracellular oxidants and increased expression of cytokines. Moreover, astrocytes incubated with the different species induced hippocampal neuronal death in co-culture, and cytotoxicity was particularly enhanced by exposure to fibrillar α-syn. Further exploration of the mechanisms behind astrocyte activation and cytotoxicity revealed differences between the assessed α-syn species. Only oligomers induced mitochondrial dysfunction in astrocytes and significantly increased extracellular hydrogen peroxide production by these cells. Besides, TNF-α and IL-1β (interleukin 1β) expression presented different kinetics and levels depending on which species induced the response. Our data suggest that α-syn species (monomeric, oligomeric and fibrillar) induce astrocyte activation that can lead to neuronal death. Nevertheless, the tested α-syn species act through different preferential mechanisms and potency. All together these results help to understand the effect of α-syn species on astrocyte function and their potential impact on the pathogenesis of Parkinson's disease and related α-synucleinopathies.
CD200 modulates spinal cord injury neuroinflammation and outcome through CD200R1
Brain Behav Immun 2018 73:416-426
Natalia Lago 1 , Bruno Pannunzio 2 , Jesús Amo-Aparicio 3 , Rubèn López-Vales 3 , Hugo Peluffo 2
1 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Mataojo 2020, 11400 Montevideo, Uruguay. Electronic address: nlago@pasteur.edu.uy. 2 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Mataojo 2020, 11400 Montevideo, Uruguay; Department of Histology and Embryology, Faculty of Medicine, UDELAR, Montevideo, Uruguay. 3 Departament de Biologia Cel·lular, Fisiologia i Immunologia, Institut de Neurociències, Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Universitat Autònoma de Barcelona, 08193 Bellaterra, Catalonia, Spain.
DOI: 10.1016/j.bbi.2018.06.002
PMID: 29870752
Pubmed: https://pubmed.ncbi.nlm.nih.gov/29870752
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S0889-1591(18)30218-6
Abstract:
The interaction between CD200 and its receptor CD200R1 is among the central regulators of microglia and macrophage phenotype. However, it remains to be established whether, in the context of a traumatic CNS injury, CD200R1 act as a negative regulator of these particular innate immune cells, and if the exogenous delivery of CD200 may ameliorate neurological deficits. In the present study, we first evaluated whether preventing the local interaction between the pair CD200-CD200R1, by using a selective blocking antibody against CD200R1, has a role on functional and inflammatory outcome after contusion-induced spinal cord injury (SCI) in mice. The injection of the αCD200R1, but not control IgG1, into the lesioned spinal cord immediately after the SCI worsened locomotor performance and exacerbated neuronal loss and demyelination. At the neuroimmunological level, we observed that microglial cells and macrophages showed increased levels of iNOS and Ly6C upon CD200R1 blockade, indicating that the disruption of CD200R1 drove these cells towards a more pro-inflammatory phenotype. Moreover, although CD200R1 blockade had no effect in the initial infiltration of neutrophils into the lesioned spinal cord, it significantly impaired their clearance, which is a key sign of excessive inflammation. Interestingly, intraparenchymal injection of recombinant CD200-His immediately after the injury induced neuroprotection and robust and long-lasting locomotor recovery. In conclusion, this study reveals that interaction of CD200-CD200R1 plays a crucial role in limiting inflammation and lesion progression after SCI, and that boosting the stimulation of this pathway may constitute a new therapeutic approach.
Profile of Arachidonic Acid-Derived Inflammatory Markers and Its Modulation by Nitro-Oleic Acid in an Inherited Model of Amyotrophic Lateral Sclerosis
Front Mol Neurosci 2018 11:131
Andrés Trostchansky 1 2 , Mauricio Mastrogiovanni 1 2 , Ernesto Miquel 2 3 , Sebastián Rodríguez-Bottero 2 3 , Laura Martínez-Palma 2 3 , Patricia Cassina 2 3 , Homero Rubbo 1 2
1 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Center for Free Radical and Biomedical Research, Universidad de la República, Montevideo, Uruguay. 3 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
DOI: 10.3389/fnmol.2018.00131
PMID: 29760648
Pubmed: https://pubmed.ncbi.nlm.nih.gov/29760648
Texto completo: https://doi.org/10.3389/fnmol.2018.00131
Abstract:
The lack of current treatments for amyotrophic lateral sclerosis (ALS) highlights the need of a comprehensive understanding of the biological mechanisms of the disease. A consistent neuropathological feature of ALS is the extensive inflammation around motor neurons and axonal degeneration, evidenced by accumulation of reactive astrocytes and activated microglia. Final products of inflammatory processes may be detected as a screening tool to identify treatment response. Herein, we focus on (a) detection of arachidonic acid (AA) metabolization products by lipoxygenase (LOX) and prostaglandin endoperoxide H synthase in SOD1G93A mice and (b) evaluate its response to the electrophilic nitro-oleic acid (NO2-OA). Regarding LOX-derived products, a significant increase in 12-hydroxyeicosatetraenoic acid (12-HETE) levels was detected in SOD1G93A mice both in plasma and brain whereas no changes were observed in age-matched non-Tg mice at the onset of motor symptoms (90 days-old). In addition, 15-hydroxyeicosatetraenoic acid (15-HETE) levels were greater in SOD1G93A brains compared to non-Tg. Prostaglandin levels were also increased at day 90 in plasma from SOD1G93A compared to non-Tg being similar in both types of animals at later stages of the disease. Administration of NO2-OA 16 mg/kg, subcutaneously (s/c) three times a week to SOD1G93A female mice, lowered the observed increase in brain 12-HETE levels compared to the non-nitrated fatty acid condition, and modified many others inflammatory markers. In addition, NO2-OA significantly improved grip strength and rotarod performance compared to vehicle or OA treated animals. These beneficial effects were associated with increased hemeoxygenase 1 (HO-1) expression in the spinal cord of treated mice co-localized with reactive astrocytes. Furthermore, significant levels of NO2-OA were detected in brain and spinal cord from NO2-OA -treated mice indicating that nitro-fatty acids (NFA) cross brain-blood barrier and reach the central nervous system to induce neuroprotective actions. In summary, we demonstrate that LOX-derived oxidation products correlate with disease progression. Overall, we are proposing that key inflammatory mediators of AA-derived pathways may be useful as novel footprints of ALS onset and progression as well as NO2-OA as a promising therapeutic compound.
Thy1-YFP-H Mice and the Parallel Rod Floor Test to Evaluate Short- and Long-Term Progression of Traumatic Brain Injury
Curr Protoc Immunol 2018 120:24.1.1-24.1.25
Monique Richter 1 2 , María Luciana Negro-Demontel 3 4 , Daniela Blanco-Ocampo 5 , Eliseo Taranto 5 , Natalia Lago 3 , Hugo Peluffo 3 4
1 Neurodegeneration Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 2 Current Address: Roche Diagnostics GmbH, Penzberg, Germany. 3 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 4 Department of Histology and Embryology, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay. 5 Department of Physiopathology, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay.
DOI: 10.1002/cpim.42
PMID: 29512144
Pubmed: https://pubmed.ncbi.nlm.nih.gov/29512144
Texto completo: https://doi.org/10.1002/cpim.42
Abstract:
Traumatic brain injury (TBI) is a leading cause of death and disability and is a risk factor for the later development of neuropsychiatric disorders and neurodegenerative diseases. Many models of TBI have been developed, but their further refinement and a more detailed long-term follow-up is needed. We have used the Thy1-YFP-H transgenic mouse line and the parallel rod floor test to produce an unbiased and robust method for the evaluation of the multiple effects of a validated model of controlled cortical injury. This approach reveals short- and long-term progressive changes, including compromised biphasic motor function up to 85 days post-lesion, which correlates with neuronal atrophy, dendrite and spine loss, and long-term axonal pathology evidenced by axon spheroids and fragmentation. Here we present methods for inducing a controlled cortical injury in the Thy1-YFP-H transgenic mouse line and for evaluating the resulting deficits in the parallel rod floor test. This technique constitutes a new, unbiased, and robust method for the evaluation of motor and behavioral alterations after TBI. © 2018 by John Wiley & Sons, Inc.
Kinesin 1 regulates cilia length through an interaction with the Bardet-Biedl syndrome related protein CCDC28B
Sci Rep 2018 8(1):3019
Rossina Novas 1 , Magdalena Cardenas-Rodriguez 1 , Paola Lepanto 1 , Matías Fabregat 1 , Magela Rodao 2 , María Inés Fariello 3 4 , Mauricio Ramos 5 , Camila Davison 2 , Gabriela Casanova 2 , Lucía Alfaya 6 , Federico Lecumberry 3 5 , Gualberto González-Sapienza 6 , Florencia Irigoín 1 7 , Jose L Badano 8
1 Human Molecular Genetics Laboratory, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, CP11400, Uruguay. 2 Unidad de Microscopía Electrónica, Facultad de Ciencias, Iguá 4225, Montevideo, CP11400, Uruguay. 3 Facultad de Ingeniería, Universidad de la República, Julio Herrera y Reissig 565, Montevideo, CP11300, Uruguay. 4 Bioinformatics Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay. 5 Signal Processing Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 6 Cátedra de Inmunología, DEPBIO, Facultad de Química, Universidad de la República, Av. Alfredo Navarro 3051, Montevideo, Uruguay. 7 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Gral. Flores 2125, Montevideo, CP11800, Uruguay. 8 Human Molecular Genetics Laboratory, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, CP11400, Uruguay. jbadano@pasteur.edu.uy.
DOI: 10.1038/s41598-018-21329-6
PMID: 29445114
Pubmed: https://pubmed.ncbi.nlm.nih.gov/29445114
Texto completo: https://doi.org/10.1038/s41598-018-21329-6
Abstract:
Bardet-Biedl syndrome (BBS) is a ciliopathy characterized by retinal degeneration, obesity, polydactyly, renal disease and mental retardation. CCDC28B is a BBS-associated protein that we have previously shown plays a role in cilia length regulation whereby its depletion results in shortened cilia both in cells and Danio rerio (zebrafish). At least part of that role is achieved by its interaction with the mTORC2 component SIN1, but the mechanistic details of this interaction and/or additional functions that CCDC28B might play in the context of cilia remain poorly understood. Here we uncover a novel interaction between CCDC28B and the kinesin 1 molecular motor that is relevant to cilia. CCDC28B interacts with kinesin light chain 1 (KLC1) and the heavy chain KIF5B. Notably, depletion of these kinesin 1 components results in abnormally elongated cilia. Furthermore, through genetic interaction studies we demonstrate that kinesin 1 regulates ciliogenesis through CCDC28B. We show that kinesin 1 regulates the subcellular distribution of CCDC28B, unexpectedly, inhibiting its nuclear accumulation, and a ccdc28b mutant missing a nuclear localization motif fails to rescue the phenotype in zebrafish morphant embryos. Therefore, we uncover a previously unknown role of kinesin 1 in cilia length regulation that relies on the BBS related protein CCDC28B.
A Rapid and Efficient Method to Dissect Pupal Wings of Drosophila Suitable for Immunodetections or PCR Assays
J Vis Exp 2017 (130):55854
Carmen Bolatto 1 , Cristina Parada 2 , Victoria Colmenares 2
1 Departamento de Histología y Embriología, Facultad de Medicina, Laboratorio de Biología del Desarrollo; cbolatto@fmed.edu.uy. 2 Departamento de Histología y Embriología, Facultad de Medicina, Laboratorio de Biología del Desarrollo.
DOI: 10.3791/55854
PMID: 29364201
Pubmed: https://pubmed.ncbi.nlm.nih.gov/29364201
Texto completo: https://doi.org//10.3791/55854
Abstract:
Wing development in Drosophila melanogaster is an ideal model for studying morphogenesis at tissue level. These appendages develop from a group of cells named wing imaginal discs formed during embryonic development. In the larval stages the imaginal discs grow, increasing its number of cells and forming monolayered epithelial structures. Inside the pupal case, the imaginal discs bud out and fold into bilayers along a line that becomes the future margin of the wing. During this process, the longitudinal primodia veins originate vein cells on the prospective dorsal and ventral surfaces of the wing. During the pupal stage the stripes of vein cells of each surface communicate in order to generate tight tubes; at the same time, the cross-veins begin their formation. With the help of appropriate molecular markers, it is possible to identify the major elements composing the wing during its development. For this reason, the ability to accurately detect transcripts or proteins in this structure is critical for studying their abundance and localization related to the development process of the wing. The procedure described here focuses on manipulating pupal wings, providing detailed instructions on how to dissect the wing during the pupal stage. The dissection of pupal tissue is more difficult to perform than their counterparts in third instar larvae. This is why this approach was developed, to obtain rapid and efficient high quality samples. Details of how to immunostain and mount these wing samples, to allow the visualization of proteins or cell components, are provided in the protocol. With little expertise it is possible to collect 8-10 high quality pupal wings in a short amount of time.
Effect of Specific Mutations in Cd300 Complexes Formation; Potential Implication of Cd300f in Multiple Sclerosis
Sci Rep 2017 7(1):13544
Águeda Martínez-Barriocanal 1 2 , Andrea Arcas-García 1 , Miriam Magallon-Lorenz 1 , Aroa Ejarque-Ortíz 1 , María Luciana Negro-Demontel 3 4 , Emma Comas-Casellas 1 , Simo Schwartz Jr 5 2 , Sunny Malhotra 3 , Xavier Montalban 3 , Hugo Peluffo 4 6 , Margarita Martín 7 , Manuel Comabella, Joan Sayós 8 9
1 CIBBIM-Nanomedicine-Immune Regulation and Immunotherapy Group. Institut de Recerca Vall Hebrón (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain. 2 Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBBER-BBN), Instituto de Salud Carlos III, Barcelona, Spain. 3 Servei de Neurologia-Neuroimmunologia. Centre d'Esclerosi Múltiple de Catalunya (Cemcat). Institut de Recerca Vall Hebrón (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain. 4 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur Montevideo, Montevideo, Uruguay. 5 CIBBIM-Nanomedicine-Drug Delivery and Targeting Group. Institut de Recerca Vall Hebrón (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain. 6 Department of Histology and Embryology, Faculty of Medicine, UDELAR, Montevideo, Uruguay. 7 Biochemistry Unit, Institut d'Investigacions Biomèdiques August Pi i Sunyer, Faculty of Medicine, University of Barcelona, Barcelona, Spain. 8 CIBBIM-Nanomedicine-Immune Regulation and Immunotherapy Group. Institut de Recerca Vall Hebrón (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain. joan.sayos@vhir.org. 9 Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBBER-BBN), Instituto de Salud Carlos III, Barcelona, Spain. joan.sayos@vhir.org.
DOI: 10.1038/s41598-017-12881-8
PMID: 29051512
Pubmed: https://pubmed.ncbi.nlm.nih.gov/29051512
Texto completo: https://doi.org/10.1038/s41598-017-12881-8
Abstract:
Herein, we have used bioinformatics tools to predict five clusters defining ligand-binding sites on the extracellular domain of human CD300b receptor, presumably involved in the formation of both homodimers and heterodimers with other CD300 family members. Site-directed mutagenesis revealed residues glutamic acid 28 and glutamine 29 in cluster 5 to be necessary for the formation of CD300b complexes. Surprisingly, the disruption of cluster 2 and 4 reconstituted the binding capability lost by the mutation of residues glutamic acid 28 to alanine, glutamine 29 to alanine (E28A-Q29G). We identified a missense mutation arginine 33 to glutamine (R33Q) in CD300f by direct sequencing of exon 2 in peripheral blood samples from 50 patients with multiple sclerosis (MS). Levels of expression of CD300f were almost undetectable on monocytes from the patient bearing the R33Q mutation compared with healthy individuals. Whereas R33Q mutation had no effect in the formation of CD300f complexes, the inhibition of protein synthesis with cycloheximide indicated that CD300f R33Q is less stable than native CD300f. Finally, we report that the levels of expression of CD300f on the surface of classical and intermediate monocytes from MS patients are significantly lower when compared to the same cell populations in healthy individuals.
Stem cells distribution, cellular proliferation and migration in the adult Austrolebias charrua brain
Brain Res 2017 1673:11-22
Maximiliano Torres-Pérez 1 , Juan Carlos Rosillo 2 , Ines Berrosteguieta 3 , Silvia Olivera-Bravo 4 , Gabriela Casanova 5 , José Manuel García-Verdugo 6 , Anabel Sonia Fernández 7
1 Departamento NCIC, Neuroanatomía Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay. Electronic address: mtorres@iibce.edu.uy. 2 Departamento NCIC, Neuroanatomía Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay; Departamento de Histología y Embriología de la Facultad de Medicina UdelaR, Avda. General Flores 2125, 11800 Montevideo, Uruguay. Electronic address: jrosillo@iibce.edu.uy. 3 Departamento NCIC, Neuroanatomía Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay. 4 Neurobiología Celular y Molecular, Instituto de Investigaciones Biológicas "Clemente Estable" (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay. Electronic address: solivera@iibce.edu.uy. 5 Unidad de Microscopia Electrónica, Facultad de Ciencias, Universidad de la República (UdelaR), Iguá 4225, 11400 Montevideo, Uruguay. Electronic address: casanova@fcien.edu.uy. 6 Laboratorio de Neurobiología Comparada, Instituto Cavanilles, Universidad de Valencia, Paterna 46980, CIBERNED, Spain. Electronic address: j.manuel.garcia@uv.es. 7 Departamento NCIC, Neuroanatomía Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay; Neuroanatomía Comparada, Unidad Asociada a la Facultad de Ciencias, Universidad de la República (UdelaR), Iguá 4225, 11400 Montevideo, Uruguay. Electronic address: anabelsonia@gmail.com.
DOI: 10.1016/j.brainres.2017.08.003
PMID: 28797690
Pubmed: https://pubmed.ncbi.nlm.nih.gov/28797690
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S0006-8993(17)30330-X
Abstract:
Our previous studies demonstrated that Austrolebias charrua annual fish is an excellent model to study adult brain cell proliferation and neurogenesis due to the presence of active and fast neurogenesis in several regions during its short lifespan. Our main goal was to identify and localize the cells that compose the neurogenic areas throughout the Austrolebias brain. To do this, we used two thymidine halogenated analogs to detect cell proliferation at different survival times: 5-chloro-2'-deoxyuridine (CldU) at 1day and 5-iodo-2'-deoxyuridine (IdU) at 30days. Three types of proliferating cells were identified: I - transient amplifying or fast cycling cells that uptake CldU; II - stem cells or slow cycling cells, that were labeled with both CldU and IdU and did not migrate; and III - migrant cells that uptake IdU. Mapping and 3D-reconstruction of labeled nuclei showed that type I and type II cells were preferentially found close to ventricle walls. Type III cells appeared widespread and migrating in tangential and radial routes. Use of proliferation markers together with Vimentin or Nestin evidenced that type II cells are the putative stem cells that are located at the ventricular lumen. Double label cells with IdU+ and NeuN or HuC/D allowed us identify migrant neurons. Quantitation of labeled nuclei indicates that the proportion of putative stem cells is around 10% in all regions of the brain. This percentage of stem cells suggests the existence of a constant brain cell population in Austrolebias charrua that seems functional to the maintainance of adult neurogenesis.