Publicaciones por año
Publicaciones del Departamento de Histología y Embriología
Neonatal testosterone exposure induces early development of follicular cysts followed by sympathetic ovarian hyperinnervation
Reprod Fertil Dev 2015 28(11) 1753-1761
Gabriel Anesetti, Rebeca Chávez-Genaro
Histology and Embryology Department, School of Medicine, General Flores 2125, CP 11800, Montevideo, Uruguay
DOI: 10.1071/RD14460
PMID: 25989716
Pubmed: https://pubmed.ncbi.nlm.nih.gov/25989716
Texto completo: https://www.publish.csiro.au/rd/RD14460
Abstract:
This study analysed the temporal association between ovarian cyst development induced by neonatal androgenisation and sympathetic innervation. Neonatal rats (postnatal Days 1 to 5) were treated with testosterone or dihydrotestosterone and the effects were evaluated at postnatal Days 20, 40, 90 or 180. Ovulation rate, number of cystic follicles and density of sympathetic fibres were analysed. The effects of surgical denervation or gonadotrophin stimulation were also assessed. Rats exposed to testosterone showed no oestrous cycle activity and did not ovulate, maintaining a polycystic ovarian morphology at all ages studied. Also, a significant increase in ovarian density of noradrenergic fibres was detected at postnatal Days 90 and 180. Sympathectomy was unable to re-establish ovarian activity; however, human chorionic gonadotrophin stimulation was enough to induce ovulation. The impact of dihydrotestosterone on ovarian function was less noticeable, showing the coexistence of corpora lutea and cystic structures without changes in sympathetic innervation. Our findings suggest that a remodelling of ovarian sympathetic innervation occurs as a response to modifications in the pattern of follicular growth induced by testosterone. A role of sympathetic innervation in the maintenance of the polycystic condition is suggested.
The Receptor CMRF35-Like Molecule-1 (CLM-1) Enhances the Production of LPS-Induced Pro-Inflammatory Mediators during Microglial Activation
PLoS One 2015 10(4):e0123928
Aroa Ejarque-Ortiz 1 , Carme Solà 2 , Águeda Martínez-Barriocanal 1 , Simó Schwartz Jr 3 , Margarita Martín 4 , Hugo Peluffo 5 , Joan Sayós 1
1 Immunobiology Group, CIBBIM-Nanomedicine Program, Hospital Universitari Vall d'Hebrón, Institut de Recerca (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBBER-BBN), Instituto de Salud Carlos III, Barcelona, Spain. 2 Department of Cerebral Ischemia and Neurodegeneration, Institut d'Investigacions Biomèdiques de Barcelona-Consejo Superior de Investigaciones Científicas (CSIC), Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain. 3 Drug Delivery and Targeting Group, CIBBIM-Nanomedicine Program, Hospital Universitari Vall d'Hebrón, Institut de Recerca (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBBER-BBN), Instituto de Salud Carlos III, Barcelona, Spain. 4 Biochemistry Unit, Faculty of Medicine, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain. 5 Neurodegeneration Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay; Department of Histology and Embryology, Faculty of Medicine, UDELAR, Montevideo, Uruguay.
DOI: 10.1371/journal.pone.0123928
PMID: 25927603
Pubmed: https://pubmed.ncbi.nlm.nih.gov/25927603
Texto completo: https://dx.plos.org/10.1371/journal.pone.0123928
Abstract:
CMRF35-like molecule-1 (CLM-1) belongs to a receptor family mainly expressed in myeloid cells that include activating and inhibitory receptors. CLM-1 contains two ITIMs and a single immunoreceptor tyrosine-based switch motif (ITSM), although also displays a binding site for p85α regulatory subunit of PI3K. By using murine primary microglial cultures, we show the presence of all CLM members in microglial cells and characterize the expression of CLM-1 both in basal conditions and during microglial activation. The TLR4 agonist lipopolysaccharide (LPS) and the TLR3 agonist polyinosinic-polycytidylic acid (Poly I:C) induce an increase in microglial CLM-1 mRNA levels in vitro, whereas the TLR2/6 heterodimer agonist peptidoglycan (PGN) produces a marked decrease. In this study we also describe a new soluble isoform of CLM-1 that is detected at mRNA and protein levels in basal conditions in primary microglial cultures. Interestingly, CLM-1 engagement enhances the transcription of the pro-inflammatory mediators TNFα, COX-2 and NOS-2 in microglial cells challenged with LPS. These results reveal that CLM-1 can acts as a co-activating receptor and suggest that this receptor could play a key role in the regulation of microglial activation.
BBB-targeting, protein-based nanomedicines for drug and nucleic acid delivery to the CNS
Biotechnol Adv 2015 33(2):277-87
Hugo Peluffo 1 , Ugutz Unzueta 2 , María Luciana Negro-Demontel 1 , Zhikun Xu 2 , Esther Váquez 2 , Neus Ferrer-Miralles 2 , Antonio Villaverde 2
1 Neuroinflammation Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay; Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República (UDELAR), Montevideo, Uruguay. 2 Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain; Department de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain; CIBER en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, 08193 Barcelona, Spain.
DOI: 10.1016/j.biotechadv.2015.02.004
PMID: 25698504
Pubmed: https://pubmed.ncbi.nlm.nih.gov/25698504
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S0734-9750(15)00029-4
Abstract:
The increasing incidence of diseases affecting the central nervous system (CNS) demands the urgent development of efficient drugs. While many of these medicines are already available, the Blood Brain Barrier and to a lesser extent, the Blood Spinal Cord Barrier pose physical and biological limitations to their diffusion to reach target tissues. Therefore, efforts are needed not only to address drug development but specially to design suitable vehicles for delivery into the CNS through systemic administration. In the context of the functional and structural versatility of proteins, recent advances in their biological fabrication and a better comprehension of the physiology of the CNS offer a plethora of opportunities for the construction and tailoring of plain nanoconjugates and of more complex nanosized vehicles able to cross these barriers. We revise here how the engineering of functional proteins offers drug delivery tools for specific CNS diseases and more transversally, how proteins can be engineered into smart nanoparticles or 'artificial viruses' to afford therapeutic requirements through alternative administration routes.
Morphological evaluation of sperm from infertile men selected by magnetic activated cell sorting (MACS)
Reprod Biol 2014 14(4):289-92
Gianni Curti 1 , Fernanda Skowronek 2 , Rita Vernochi 3 , Ana Laura Rodriguez-Buzzi 4 , Juan Carlos Rodriguez-Buzzi 5 , Gabriela Casanova 6 , Rossana Sapiro 7
1 Departamento de Biologia Molecular - Instituto De Investigaciones Biológicas Clemente Estable (IIBCE), Avenida Italia 3318, CP 11600 Montevideo, Uruguay. Electronic address: giannicurti@gmail.com. 2 Laboratorio de Biología Molecular de la Reproducción - Departamento de Histología y Embriología, Facultad de Medicina, Gral. Flores 2125, 11800 Montevideo, Uruguay. Electronic address: fernandaskowronek@gmail.com. 3 Centro de Reproducción Humana del Interior (CERHIN), Bvar. Artigas 1665/102, Montevideo, Uruguay. Electronic address: draritaverno@gmail.com. 4 Centro de Reproducción Humana del Interior (CERHIN), Bvar. Artigas 1665/102, Montevideo, Uruguay. Electronic address: anucharod@gmail.com. 5 Centro de Reproducción Humana del Interior (CERHIN), Bvar. Artigas 1665/102, Montevideo, Uruguay. Electronic address: jcrbuzzi@gmail.com. 6 Unidad de Microscopía Electrónica de Transmisión - Facultad de Ciencias, Iguá 4225, 11400 Montevideo, Uruguay. Electronic address: paragaby@gmail.com. 7 Laboratorio de Biología Molecular de la Reproducción - Departamento de Histología y Embriología, Facultad de Medicina, Gral. Flores 2125, 11800 Montevideo, Uruguay. Electronic address: rsapiro@fmed.edu.uy.
DOI: 10.1016/j.repbio.2014.07.002
PMID: 25454495
Pubmed: https://pubmed.ncbi.nlm.nih.gov/25454495
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S1642-431X(14)00074-6
Abstract:
Electron microscopy analysis performed in five infertile human subjects after sperm selection by swim-up followed by magnetic activated cell sorting (MACS) demonstrated a decrease in the number of spermatozoa with characteristics compatible with cell death. However, no significant differences were found when the swim-up/MACS semen fraction was compared with swim-up fraction alone.
Comparative analysis of lentiviral vectors and modular protein nanovectors for traumatic brain injury gene therapy
ol Ther Methods Clin Dev 2014 1:14047.
María Luciana Negro-Demontel 1 , Paolo Saccardo 2 , Cecilia Giacomini 3 , Rafael Joaquín Yáñez-Muñoz 4 , Neus Ferrer-Miralles 2 , Esther Vazquez 2 , Antonio Villaverde 2 , Hugo Peluffo 1
1 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo , Montevideo, Uruguay ; Departmento de Histología y Embriología, Facultad de Medicina, UDELAR , Montevideo, Uruguay. 2 Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona , Barcelona, Spain ; Department de Genètica i de Microbiologia, Universitat Autònoma de Barcelona , Barcelona, Spain ; CIBER en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN) , Barcelona, Spain. 3 Cátedra de Bioquímica, Departamento de Biociencias, Facultad de Química, UDELAR , Montevideo, Uruguay. 4 Royal Holloway University of London , Egham, UK.
DOI: 10.1038/mtm.2014.47
PMID: 26015985
Pubmed: https://pubmed.ncbi.nlm.nih.gov/26015985
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S2329-0501(16)30115-2
Abstract:
Traumatic brain injury (TBI) remains as one of the leading causes of mortality and morbidity worldwide and there are no effective treatments currently available. Gene therapy applications have emerged as important alternatives for the treatment of diverse nervous system injuries. New strategies are evolving with the notion that each particular pathological condition may require a specific vector. Moreover, the lack of detailed comparative studies between different vectors under similar conditions hampers the selection of an ideal vector for a given pathological condition. The potential use of lentiviral vectors versus several modular protein-based nanovectors was compared using a controlled cortical impact model of TBI under the same gene therapy conditions. We show that variables such as protein/DNA ratio, incubation volume, and presence of serum or chloroquine in the transfection medium impact on both nanovector formation and transfection efficiency in vitro. While lentiviral vectors showed GFP protein 1 day after TBI and increased expression at 14 days, nanovectors showed stable and lower GFP transgene expression from 1 to 14 days. No toxicity after TBI by any of the vectors was observed as determined by resulting levels of IL-1β or using neurological sticky tape test. In fact, both vector types induced functional improvement per se.
Neuroprotective effects of the mitochondria-targeted antioxidant MitoQ in a model of inherited amyotrophic lateral sclerosis
Free Radic Biol Med 2014 70:204-13
Ernesto Miquel 1 , Adriana Cassina 2 , Laura Martínez-Palma 1 , José M Souza 2 , Carmen Bolatto 1 , Sebastián Rodríguez-Bottero 1 , Angela Logan 3 , Robin A J Smith 4 , Michael P Murphy 3 , Luis Barbeito 5 , Rafael Radi 2 , Patricia Cassina 6
1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay. 2 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay; Center for Free Radical and Biomedical Research, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay. 3 Mitochondrial Biology Unit, Medical Research Council, Cambridge CB2 0XY, UK. 4 Department of Chemistry, University of Otago, Dunedin 9054, New Zealand. 5 Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay. 6 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay; Center for Free Radical and Biomedical Research, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay
DOI: 10.1016/j.freeradbiomed.2014.02.019
PMID: 24582549
Pubmed: https://pubmed.ncbi.nlm.nih.gov/24582549
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S0891-5849(14)00096-3
Abstract:
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by motor neuron degeneration that ultimately results in progressive paralysis and death. Growing evidence indicates that mitochondrial dysfunction and oxidative stress contribute to motor neuron degeneration in ALS. To further explore the hypothesis that mitochondrial dysfunction and nitroxidative stress contribute to disease pathogenesis at the in vivo level, we assessed whether the mitochondria-targeted antioxidant [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methane sulfonate (MitoQ) can modify disease progression in the SOD1(G93A) mouse model of ALS. To do this, we administered MitoQ (500 µM) in the drinking water of SOD1(G93A) mice from a time when early symptoms of neurodegeneration become evident at 90 days of age until death. This regime is a clinically plausible scenario and could be more easily translated to patients as this corresponds to initiating treatment of patients after they are first diagnosed with ALS. MitoQ was detected in all tested tissues by liquid chromatography/mass spectrometry after 20 days of administration. MitoQ treatment slowed the decline of mitochondrial function, in both the spinal cord and the quadriceps muscle, as measured by high-resolution respirometry. Importantly, nitroxidative markers and pathological signs in the spinal cord of MitoQ-treated animals were markedly reduced and neuromuscular junctions were recovered associated with a significant increase in hindlimb strength. Finally, MitoQ treatment significantly prolonged the life span of SOD1(G93A) mice. Our results support a role for mitochondrial nitroxidative damage and dysfunction in the pathogenesis of ALS and suggest that mitochondria-targeted antioxidants may be of pharmacological use for ALS treatment.
Pharmacological study of the one spike spherical neuron phenotype in Gymnotus omarorum
Neuroscience 2014 258:347-54
J Nogueira 1, A A Caputi 2
1 Department of Integrative and Computational Neurosciences, Instituto de Investigaciones Biológicas Clemente Estable, Avenue Italia 3318, Montevideo, Uruguay; Department of Histology and Embriology, Facultad de Medicina, Universidad de la República, Gral. Flores 2515, Montevideo, Uruguay. 2 Department of Integrative and Computational Neurosciences, Instituto de Investigaciones Biológicas Clemente Estable, Avenue Italia 3318, Montevideo, Uruguay
DOI: 10.1016/j.neuroscience.2013.11.021
PMID: 24269939
Pubmed: https://pubmed.ncbi.nlm.nih.gov/24269939
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S0306-4522(13)00961-5
Abstract:
The intrinsic properties of spherical neurons play a fundamental role in the sensory processing of self-generated signals along a fast electrosensory pathway in electric fish. Previous results indicate that the spherical neuron's intrinsic properties depend mainly on the presence of two resonant currents that tend to clamp the voltage near the resting potential. Here we show that these are: a low-threshold potassium current blocked by 4-aminopyridine and a mixed cationic current blocked by cesium chloride. We also show that the low-threshold potassium current also causes the long refractory period, explaining the necessary properties that implement the dynamic filtering of the self-generated signals previously described. Comparative data from other fish and from the auditory system indicate that other single spiking onset neurons might differ in the channel repertoire observed in the spherical neurons of Gymnotus omarorum.
From the intrinsic properties to the functional role of a neuron phenotype: an example from electric fish during signal trade-off
J Exp Bio 2013 216(Pt 13):2380-92
Javier Nogueira 1 2, Angel A Caputi 2
1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Avenida General Flores, 2125 Montevideo, Uruguay 2 Departamento de Neurociencias Integrativas y Computacionales, Instituto de Investigaciones Biológicas Clemente Estable, Avenida Italia, 3318 Montevideo, Uruguay
DOI: 10.1242/jeb.082651
PMID: 23761463
Pubmed: https://pubmed.ncbi.nlm.nih.gov/23761463
Texto completo: https://journals.biologists.com/jeb/article-lookup/doi/10.1242/jeb.082651
Abstract:
This review deals with the question: what is the relationship between the properties of a neuron and the role that the neuron plays within a given neural circuit? Answering this kind of question requires collecting evidence from multiple neuron phenotypes and comparing the role of each type in circuits that perform well-defined computational tasks. The focus here is on the spherical neurons in the electrosensory lobe of the electric fish Gymnotus omarorum. They belong to the one-spike-onset phenotype expressed at the early stages of signal processing in various sensory modalities and diverse taxa. First, we refer to the one-spike neuron intrinsic properties, their foundation on a low-threshold K(+) conductance, and the potential roles of this phenotype in different circuits within a comparative framework. Second, we present a brief description of the active electric sense of weakly electric fish and the particularities of spherical one-spike-onset neurons in the electrosensory lobe of G. omarorum. Third, we introduce one of the specific tasks in which these neurons are involved: the trade-off between self- and allo-generated signals. Fourth, we discuss recent evidence indicating a still-undescribed role for the one-spike phenotype. This role deals with the blockage of the pathway after being activated by the self-generated electric organ discharge and how this blockage favors self-generated electrosensory information in the context of allo-generated interference. Based on comparative analysis we conclude that one-spike-onset neurons may play several functional roles in animal sensory behavior. There are specific adaptations of the neuron's 'response function' to the circuit and task. Conversely, the way in which a task is accomplished depends on the intrinsic properties of the neurons involved. In short, the role of a neuron within a circuit depends on the neuron and its functional context.
Overexpression of the nuclear factor kappaB inhibitor A20 is neurotoxic after an excitotoxic injury to the immature rat brain
Neurol Res 2013 35(3):308-19
Hugo Peluffo 1 2, Pau Gonzalez 3, Laia Acarin 4, Anna Arís 5, Rudy Beyaert 6 7, Antonio Villaverde 5 8, Berta Gonzalez 4
1 Laboratorio de Neurodegeneración, Institut Pasteur de Montevideo, Uruguay 2 Laboratorio de Neurobiología Celular y Molecular, Depto. Histología y Embriología, Facultad de Medicina, UDELAR, Uruguay, 3 Laboratorio de Neurología Molecular, Hospital Nacional de Parapléjicos de Toledo, Spain, 4 Unitat d’Histologia, Torre M5, Facultat de Medicina, Departament de Biologia Cellular, Fisiologia i Immunologia, and Institut de Neurociéncies, Universitat Autònoma de Barcelona, Spain, 5 Institut de Biotecnologia i de Biomedicina and Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Spain, 6 Department of Molecular Biomedical Research, Unit of Molecular Signal Transduction in Inflammation, VIB, Belgium, 7 Department of Molecular Biology, Ghent University, Belgium, 8CIBER de Bioingenierı ´a, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, Barcelona, Spain
DOI: 10.1179/1743132812Y.0000000139
PMID: 23336395
Pubmed: https://pubmed.ncbi.nlm.nih.gov/23336395
Texto completo: https://www.tandfonline.com/doi/full/10.1179/1743132812Y.0000000139
Abstract:
Background: The zinc finger protein A20 is an ubiquitinating/deubiquitinating enzyme essential for the termination of inflammatory reactions through the inhibition of nuclear factor kappaB (NF-kappaB) signaling. Moreover, it also shows anti-apoptotic activities in some cell types and proapoptotic/pronecrotic effects in others. Although it is known that the regulation of inflammatory and cell death processes are critical in proper brain functioning and that A20 mRNA is expressed in the CNS, its role in the brain under physiological and pathological conditions is still unknown.
Methods: In the present study, we have evaluated the effects of A20 overexpression in mixed cortical cultures in basal conditions: the in vivo pattern of endogenous A20 expression in the control and N-methyl-d-aspartate (NMDA) excitotoxically damaged postnatal day 9 immature rat brain, and the post-injury effects of A20 overexpression in the same lesion model.
Results: Our results show that overexpression of A20 in mixed cortical cultures induced significant neuronal death by decreasing neuronal cell counts by 45 ± 9%. in vivo analysis of endogenous A20 expression showed widespread expression in gray matter, mainly in neuronal cells. However, after NMDA-induced excitotoxicity, neuronal A20 was downregulated in the neurodegenerating cortex and striatum at 10-24 hours post-lesion, and it was re-expressed at longer survival times in reactive astrocytes located mainly in the lesion border. When A20 was overexpressed in vivo 2 hours after the excitotoxic damage, the lesion volume at 3 days post-lesion showed a significant increase (20.8 ± 7.0%). No A20-induced changes were observed in the astroglial response to injury.
Conclusions: A20 is found in neuronal cells in normal conditions and is also expressed in astrocytes after brain damage, and its overexpression is neurotoxic for cortical neurons in basal mixed neuron-glia culture conditions and exacerbates postnatal brain excitotoxic damage.
Convergence of multiple mechanisms of steroid hormone action
Horm Metab Res 2012 44(8):569-76
S K Mani 1 *, P G Mermelstein 2 *, M J Tetel 3 *, G Anesetti 4 *
1 Department of Molecular & Cellular Biology and Neuroscience, Baylor College of Medicine, Houston, TX, USA 2 Department of Neuroscience, University of Minnesota, Minneapolis, MN, USA 3 Neuroscience Program, Wellesley College, Wellesley, MA, USA 4 Departamento de Hostologia y Embriologia, Facultad de Medicine, Universidad de la Republica, Montevideo, Uruguay *All the authors contributed equally to this work.
DOI: 10.1055/s-0032-1306343
PMID: 22454239
Pubmed: https://pubmed.ncbi.nlm.nih.gov/22454239
Texto completo: https://www.thieme-connect.com/products/ejournals/abstract/10.1055/s-0032-1306343
Abstract:
Steroid hormones modulate a wide array of physiological processes including development, metabolism, and reproduction in various species. It is generally believed that these biological effects are predominantly mediated by their binding to specific intracellular receptors resulting in conformational change, dimerization, and recruitment of coregulators for transcription-dependent genomic actions (classical mechanism). In addition, to their cognate ligands, intracellular steroid receptors can also be activated in a "ligand-independent" manner by other factors including neurotransmitters. Recent studies indicate that rapid, nonclassical steroid effects involve extranuclear steroid receptors located at the membrane, which interact with cytoplasmic kinase signaling molecules and G-proteins. The current review deals with various mechanisms that function together in an integrated manner to promote hormone-dependent actions on the central and sympathetic nervous systems.