Publicaciones por año
Publicaciones del Departamento de Histología y Embriología
Mitochondrial metabolism determines the functional status of human sperm and correlates with semen parameters
Front Cell Dev Biol 2022 10:926684
Pilar Irigoyen 1 , Paula Pintos-Polasky 1 , Lucia Rosa-Villagran 1 , Maria Fernanda Skowronek 1 , Adriana Cassina 2 3 , Rossana Sapiro 1 3
1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
DOI: 10.3389/fcell.2022.926684
PMID: 36111336
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36111336
Texto completo: https://doi.org/10.3389/fcell.2022.926684
Abstract:
The diagnosis of male infertility is based essentially on the patient's medical history and a standard semen analysis. However, the latter rarely provides information on the causes of a possible infertility, emphasizing the need to extend the analysis of the sperm function. Mitochondrial function has been associated with sperm function and dysfunction, the latter primarily through the production of excessive amounts of reactive oxygen species (ROS). We hypothesized that analysis of sperm mitochondrial metabolism together with sperm ROS production could be an additional tool to improve routine semen analysis, after appropriate validations. To test our hypothesis, we performed several experiments using a non-routine method (high-resolution respirometry, HRR) to access mitochondrial function. First, we investigated whether mitochondrial function is related to human sperm motility and morphology. When mitochondrial metabolism was challenged, sperm motility decreased significantly. Additionally, morphological abnormalities in the sperm mid-piece and mitochondria were associated with global sperm defects evaluated by routine methods. Subsequently, sperm mitochondrial function was assessed by HRR. Respiratory control ratio (RCR) was determined and evaluated in the context of classical sperm analysis. In parallel, sperm hydrogen peroxide (H2O2) production and seminal plasma (SP) antioxidant capacity were measured. The percentage of sperm with progressive motility correlated positively with RCR, SP antioxidant capacity, and negatively with the concentration of extracellular H2O2 production ([H2O2]). The percentage of normal sperm morphology correlated positively with RCR and negatively with [H2O2]. Sperm morphology did not correlate with seminal plasma antioxidant capacity. Furthermore, Receiver Operating Characteristic curves were used for the first time to test the diagnostic ability of RCR, [H2O2], and SP antioxidant capacity as binary classifiers. An RCR cut off value of 3.2 was established with a sensitivity of 73% and a specificity of 61%, using reference values considered normal or abnormal in routine semen analysis. The cut off value for [H2O2] was 0.2 μM/106 sperm (sensitivity = 65%, specificity = 60%). There were no reference values for SP antioxidant capacity that distinguished between abnormal and normal sperm samples. We conclude that sperm mitochondrial function indices in combination with [H2O2] may be useful tools to complement the routine semen analysis.
The application of artificial gravity in medicine and space
Front Physiol 2022 13:952723
Eugenia Isasi 1 2 , Maria E Isasi 1 , Jack J W A van Loon 3 4
1 Centro de Terapia Gravitacional, Montevideo, Uruguay. 2 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Department of Oral and Maxillofacial Surgery/Pathology, Amsterdam Movement Sciences & Amsterdam Bone Center (ABC), Amsterdam UMC location Vrije Universiteit Amsterdam & Academic Center for Dentistry Amsterdam (ACTA), Amsterdam, Netherlands. 4 Life Support and Physical Sciences Section (TEC-MMG), European Space Agency (ESA), European Space Research and Technology Centre (ESTEC), Noordwijk, Netherlands.
DOI: 10.3389/fphys.2022.952723
PMID: 36105282
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36105282
Texto completo: https://doi.org/10.3389/fphys.2022.952723
Abstract:
Gravity plays a crucial role in physiology. The lack of gravity, like in long duration spaceflight missions, cause pathologies in e.g., the musculoskeletal system, cardiovascular deconditioning, immune system deprivation or brain abnormalities, to just mention a few. The application of artificial gravity through short-arm human centrifugation (SAHC) has been studied as a possible countermeasure to treat spaceflight deconditioning. However, hypergravity protocols applied by using SAHC have also been used to treat different, ground-based pathologies. Such gravitational therapies have been applied in Uruguay for more than four decades now. The aim of this overview is to summarize the most important findings about the effects of gravitational therapy in different, mainly vascular based pathologies according to the experience in the Gravitational Therapy Center and to discuss the current research in the field of hypergravity applications in medicine but also as multisystem countermeasure for near weightlessness pathologies. New insight is needed on the use of hypergravity in medicine and space research and application.
Patched-Related Is Required for Proper Development of Embryonic Drosophila Nervous System
Front Neurosci 2022 16:920670
Carmen Bolatto 1 2 , Sofía Nieves 1 , Agustina Reyes 1 , Silvia Olivera-Bravo 2 , Verónica Cambiazo 3
1 Developmental Biology Laboratory, Histology and Embryology Department, Faculty of Medicine, Universidad de la República (UdelaR), Montevideo, Uruguay. 2 Cell and Molecular Neurobiology Laboratory, Computational and Integrative Neuroscience (NCIC) Department, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Montevideo, Uruguay. 3 Bioinformatic and Gene Expression Laboratory, Institute of Nutrition and Food Technology (INTA)-Universidad de Chile and Millennium Institute Center for Genome Regulation (CRG), Santiago, Chile.
DOI: 10.3389/fnins.2022.920670
PMID: 36081658
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36081658
Texto completo: https://doi.org/10.3389/fnins.2022.920670
Abstract:
Patched-related (Ptr), classified primarily as a neuroectodermal gene, encodes a protein with predicted topology and domain organization closely related to those of Patched (Ptc), the canonical receptor of the Hedgehog (Hh) pathway. To investigate the physiological function of Ptr in the developing nervous system, Ptr null mutant embryos were immunolabeled and imaged under confocal microscopy. These embryos displayed severe alterations in the morphology of the primary axonal tracts, reduced number, and altered distribution of the Repo-positive glia as well as peripheral nervous system defects. Most of these alterations were recapitulated by downregulating Ptr expression, specifically in embryonic nerve cells. Because similar nervous system phenotypes have been observed in hh and ptc mutant embryos, we evaluated the Ptr participation in the Hh pathway by performing cell-based reporter assays. Clone-8 cells were transfected with Ptr-specific dsRNA or a Ptr DNA construct and assayed for changes in Hh-mediated induction of a luciferase reporter. The results obtained suggest that Ptr could act as a negative regulator of Hh signaling. Furthermore, co-immunoprecipitation assays from cell culture extracts premixed with a conditioned medium revealed a direct interaction between Ptr and Hh. Moreover, in vivo Ptr overexpression in the domain of the imaginal wing disc where Engrailed and Ptc coexist produced wing phenotypes at the A/P border. Thus, these results strongly suggest that Ptr plays a crucial role in nervous system development and appears to be a negative regulator of the Hh pathway.
Generation and characterization of Ccdc28b mutant mice links the Bardet-Biedl associated gene with mild social behavioral phenotypes
PLoS Genet 2022 18(6):e1009896
Matías Fabregat 1 2 , Sofía Niño-Rivero 3 , Sabrina Pose 4 , Magdalena Cárdenas-Rodríguez 1 2 , Mariana Bresque 2 5 , Karina Hernández 6 , Victoria Prieto-Echagüe 1 2 , Geraldine Schlapp 7 , Martina Crispo 7 , Patricia Lagos 3 , Natalia Lago 4 , Carlos Escande 2 5 , Florencia Irigoín 1 2 6 , Jose L Badano 1 2
1 Human Molecular Genetics Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 2 INDICyO Institutional Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. 3 Departamento de Fisiología, Universidad de la República, Montevideo, Uruguay. 4 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 5 Metabolic Diseases and Aging Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 6 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 7 Laboratory Animal Biotechnology Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay.
DOI: 10.1371/journal.pgen.1009896
PMID: 35653384
Pubmed: https://pubmed.ncbi.nlm.nih.gov/35653384
Texto completo: https://dx.plos.org/10.1371/journal.pgen.1009896
Abstract:
CCDC28B (coiled-coil domain-containing protein 28B) was identified as a modifier in the ciliopathy Bardet-Biedl syndrome (BBS). Our previous work in cells and zebrafish showed that CCDC28B plays a role regulating cilia length in a mechanism that is not completely understood. Here we report the generation of a Ccdc28b mutant mouse using CRISPR/Cas9 (Ccdc28b mut). Depletion of CCDC28B resulted in a mild phenotype. Ccdc28b mut animals i) do not present clear structural cilia affectation, although we did observe mild defects in cilia density and cilia length in some tissues, ii) reproduce normally, and iii) do not develop retinal degeneration or obesity, two hallmark features of reported BBS murine models. In contrast, Ccdc28b mut mice did show clear social interaction defects as well as stereotypical behaviors. This finding is indeed relevant regarding CCDC28B as a modifier of BBS since behavioral phenotypes have been documented in BBS. Overall, this work reports a novel mouse model that will be key to continue evaluating genetic interactions in BBS, deciphering the contribution of CCDC28B to modulate the presentation of BBS phenotypes. In addition, our data underscores a novel link between CCDC28B and behavioral defects, providing a novel opportunity to further our understanding of the genetic, cellular, and molecular basis of these complex phenotypes.
Plasticity of cell proliferation in the retina of Austrolebias charrua fish under light and darkness conditions
Curr Res Neurobiol 2022 3:100042
Inés Berrosteguieta 1 , Juan Carlos Rosillo 1 2 , María Laura Herrera 1 3 , Silvia Olivera-Bravo 4 , Gabriela Casanova 5 , Vicente Herranz-Pérez 6 , José Manuel García-Verdugo 6 , Anabel Sonia Fernández 1 3
1 Departamento Neurociencias Integrativas, Lab. Neurobiología Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600, Montevideo, Uruguay. 2 Departamento de Histología y Embriología, Facultad de Medicina, UdelaR. Avda. General Flores 2125, 11800, Montevideo, Uruguay. 3 Facultad de Ciencias, UdelaR, Iguá 4225, 11400, Montevideo, Uruguay. 4 Neurobiología Celular y Molecular, IIBCE, Avenida. Italia 3318, 11600, Montevideo, Uruguay. 5 Unidad de Microscopía Electrónica, Facultad de Ciencias, Universidad de la República (UdelaR), Iguá 4225, 11400, Montevideo, Uruguay. 6 Laboratorio de Neurobiología Comparada, Instituto Cavanilles, Universitat de València, CIBERNED, 46980, Paterna, Spain.
DOI: 10.1016/j.crneur.2022.100042
PMID: 36518338
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36518338
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S2665-945X(22)00015-8
Abstract:
Austrolebias annual fishes exhibit cell proliferation and neurogenesis throughout life. They withstand extreme environmental changes as their habitat dries out, pressuring nervous system to adapt. Their visual system is challenged to adjust as the water becomes turbid. Therefore, this study focused on how change in photic environment can lead to an increased cell proliferation in the retina. We administered 5-chloro-2'- deoxyuridine (CldU) and 5-iodo-2'-deoxyuridine (IdU) at different temporal windows to detect cell proliferation in natural light and permanent darkness. Stem/progenitor cells were recognized as IdU+/CldU + nuclei co-labeled with Sox2, Pax6 or BLBP found in the ciliary marginal zone (CMZ). The expression pattern of BLBP + glial cells and ultrastructural analysis indicates that CMZ has different cell progenitors. In darkness, the number of dividing cells significantly increased, compared to light conditions. Surprisingly, CMZ IdU+/CldU + cell number was similar under light and darkness, suggesting a stable pool of stem/progenitor cells possibly responsible for retinal growth. Therefore, darkness stimulated cell progenitors outside the CMZ, where Müller glia play a crucial role to generate rod precursors and other cell types that might integrate rod-dependent circuits to allow darkness adaptation. Thus, the Austrolebias fish retina shows great plasticity, with cell proliferation rates significantly higher than that of brain visual areas.
Gestational and Lactational Iron Deficiency Anemia Impairs Myelination and the Neurovascular Unit in Infant Rats
Mol Neurobiol 2022 59(6):3738-3754
Eugenia Isasi 1 2 , Martin Figares 2 3 , Verónica Abudara 3 , Silvia Olivera-Bravo 4
1 Neurobiología Celular y Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), 3318, Italia Av., 11600, Montevideo, Uruguay. 2 Neurobiología Celular y Molecular, Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Departamento de Fisiología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 4 Neurobiología Celular y Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), 3318, Italia Av., 11600, Montevideo, Uruguay. solivera2011@gmail.com.
DOI: 10.1007/s12035-022-02798-3
PMID: 35381889
Pubmed: https://pubmed.ncbi.nlm.nih.gov/35381889
Texto completo: https://dx.doi.org/10.1007/s12035-022-02798-3
Abstract:
Iron deficiency anemia is a prevalent health problem among pregnant women and infants, particularly in the developing countries that causes brain development deficits and poor cognitive outcomes. Since tissue iron depletion may impair myelination and trigger cellular hypoxic signaling affecting blood vessels, we studied myelination and the neurovascular unit (NVU) in infant rats born to mothers fed with an iron deficient (ID) or control diet from embryonic day 5 till weaning. Blood samples and brains of rat pups at postnatal day (PND) 14 and 30 were analyzed. PND 14 ID rats had severe microcytic hypochromic anemia that was almost reversed at PND 30 although hypomyelination and astrocyte immature phenotype in the corpus callosum were significant at that age. In CA1 hippocampal region, PND 14 and PND 30 ID rats showed significant reduced expression of the receptor β of the platelet-derived growth factor localized in pericytes and associated to aquaporin 4 (AQP4) immunopositive capillaries. Shorter AQP4 + capillaries and reduced AQP4 expression were also evidenced in PND 14 and PND 30 ID rats. In addition, pericyte membrane permeability through large-pore channels was transiently increased in ID rats at PND 14 but not at PND 30, while the blood-brain barrier permeability was not affected. Remarkably, transient increased pericyte permeability found in PND 14 ID rats was not directly related to iron depletion, suggesting the involvement of other iron deficiency anemia-induced mechanisms. In summary, severe ID during gestation and lactation produces persistent hypomyelination and significantly affects hippocampal pericytes and astrocytes in the NVU which may trigger impaired neurovascular function.
CD200R1 Contributes to Successful Functional Reinnervation after a Sciatic Nerve Injury
Cells 2022 11(11):1786
Bruno Pannunzio 1 2 , Jesús Amo-Aparicio 3 , Camila Julián 2 , Rubèn López-Vales 3 , Hugo Peluffo 1 2 4 , Natalia Lago 1 3
1 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay. 2 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo 11800, 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. 4 Unitat de Bioquímica i Biologia Molecular, Departament de Biomedicina, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, 08036 Barcelona, Catalonia, Spain.
DOI: 10.3390/cells11111786
PMID: 35681481
Pubmed: https://pubmed.ncbi.nlm.nih.gov/35681481
Texto completo: https://www.mdpi.com/resolver?pii=cells11111786
Abstract:
Activating and inhibitory immune receptors play a critical role in regulating systemic and central nervous system (CNS) immune and inflammatory processes. The CD200R1 immunoreceptor induces a restraining signal modulating inflammation and phagocytosis in the CNS under different inflammatory conditions. However, it remains unknown whether CD200R1 has a role in modulating the inflammatory response after a peripheral nerve injury, an essential component of the successful regeneration. Expression of CD200R1 and its ligand CD200 was analyzed during homeostasis and after a sciatic nerve crush injury in C57Bl/6 mice. The role of CD200R1 in Wallerian Degeneration (WD) and nerve regeneration was studied using a specific antibody against CD200R1 injected into the nerve at the time of injury. We found an upregulation of CD200R1 mRNA after injury whereas CD200 was downregulated acutely after nerve injury. Blockade of CD200R1 significantly reduced the acute entrance of both neutrophils and monocytes from blood after nerve injury. When long term regeneration and functional recovery were evaluated, we found that blockade of CD200R1 had a significant effect impairing the spontaneous functional recovery. Taken together, these results show that CD200R1 has a role in mounting a successful acute inflammatory reaction after injury, and contributes to an effective functional recovery.
Colocalization Analysis of Peripheral Myelin Protein-22 and Lamin-B1 in the Schwann Cell Nuclei of Wt and TrJ Mice
Biomolecules 2022 12(3):456
María Vittoria Di Tomaso 1 , Lucía Vázquez Alberdi 2 , Daniela Olsson 1 , Saira Cancela 1 , Anabel Fernández 3 4 , Juan Carlos Rosillo 3 5 , Ana Laura Reyes Ábalos 1 6 , Magdalena Álvarez Zabaleta 1 , Miguel Calero 7 8 , Alejandra Kun 2 9
1 Departamento de Genética, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo 11600, Uruguay. 2 Laboratorio de Biología Celular del Sistema Nervioso Periférico, Departamento de Proteínas y Ácidos Nucleicos, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo 11600, Uruguay. 3 Laboratorio de Neurobiología Comparada, Departamento de Neurociencias Integrativas, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo 11600, Uruguay. 4 Laboratorio de Neurociencias, Facultad de Ciencias, Universidad de la República, Montevideo 11400, Uruguay. 5 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo 11800, Uruguay. 6 Unidad de Microscopía Electrónica de Barrido, Universidad de la República, Montevideo 11400, Uruguay. 7 Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), Unidad de Encefalopatías Espongiformes (UFIEC), 28029 Madrid, Spain. 8 Queen Sofia Foundation Alzheimer Center, CIEN Foundation, 28031 Madrid, Spain. 9 Sección Bioquímica, Facultad de Ciencias, Universidad de la República, Montevideo 11400, Uruguay.
DOI: 10.3390/biom12030456
PMID: 35327648
Pubmed: https://pubmed.ncbi.nlm.nih.gov/35327648
Texto completo: https://www.mdpi.com/resolver?pii=biom12030456
Abstract:
Myelination of the peripheral nervous system requires Schwann cells (SC) differentiation into the myelinating phenotype. The peripheral myelin protein-22 (PMP22) is an integral membrane glycoprotein, expressed in SC. It was initially described as a growth arrest-specific (gas3) gene product, up-regulated by serum starvation. PMP22 mutations were pathognomonic for human hereditary peripheral neuropathies, including the Charcot-Marie-Tooth disease (CMT). Trembler-J (TrJ) is a heterozygous mouse model carrying the same pmp22 point mutation as a CMT1E variant. Mutations in lamina genes have been related to a type of peripheral (CMT2B1) or central (autosomal dominant leukodystrophy) neuropathy. We explore the presence of PMP22 and Lamin B1 in Wt and TrJ SC nuclei of sciatic nerves and the colocalization of PMP22 concerning the silent heterochromatin (HC: DAPI-dark counterstaining), the transcriptionally active euchromatin (EC), and the nuclear lamina (H3K4m3 and Lamin B1 immunostaining, respectively). The results revealed that the number of TrJ SC nuclei in sciatic nerves was greater, and the SC volumes were smaller than those of Wt. The myelin protein PMP22 and Lamin B1 were detected in Wt and TrJ SC nuclei and predominantly in peripheral nuclear regions. The level of PMP22 was higher, and those of Lamin B1 lower in TrJ than in Wt mice. The level of PMP22 was higher, and those of Lamin B1 lower in TrJ than in Wt mice. PMP22 colocalized more with Lamin B1 and with the transcriptionally competent EC, than the silent HC with differences between Wt and TrJ genotypes. The results are discussed regarding the probable nuclear role of PMP22 and the relationship with TrJ neuropathy.
Neuroprotective effects of violacein in a model of inherited amyotrophic lateral sclerosis
Sci Rep 2022 12(1):4439
Silvia Olivera-Bravo 1 , Carmen Bolatto 2 3 , Gabriel Otero Damianovich 2 , Matías Stancov 2 , Sofía Cerri 2 3 , Paola Rodríguez 2 , Daniela Boragno 2 , Karina Hernández Mir 3 , María Noel Cuitiño 2 , Fernanda Larrambembere 2 , Eugenia Isasi 2 3 , Diego Alem 4 , Lucía Canclini 4 , Marta Marco 2 5 , Danilo Davyt 6 , Pablo Díaz-Amarilla 2
1 Cell and Mol Neurobiol Lab, NCIC Department, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Montevideo, Uruguay. solivera@iibce.edu.uy. 2 Cell and Mol Neurobiol Lab, NCIC Department, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Montevideo, Uruguay. 3 Histology and Embryology Department, Faculty of Medicine, Universidad de La República (UdelaR), Montevideo, Uruguay. 4 Genetic Department, IIBCE, Montevideo, Uruguay. 5 Tumoral Biol Area, Clin Biochem Department, Faculty of Chemistry, UdelaR, Montevideo, Uruguay. 6 Pharm Chem Lab, Organic Chemistry Department, Faculty of Chemistry, UdelaR, Montevideo, Uruguay.
DOI: 10.1038/s41598-022-06470-7
PMID: 35292673
Pubmed: https://pubmed.ncbi.nlm.nih.gov/35292673
Texto completo: https://doi.org/10.1038/s41598-022-06470-7
Abstract:
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive death of motor neurons and muscle atrophy, with defective neuron-glia interplay and emergence of aberrant glial phenotypes having a role in disease pathology. Here, we have studied if the pigment violacein with several reported protective/antiproliferative properties may control highly neurotoxic astrocytes (AbAs) obtained from spinal cord cultures of symptomatic hSOD1G93A rats, and if it could be neuroprotective in this ALS experimental model. At concentrations lower than those reported as protective, violacein selectively killed aberrant astrocytes. Treatment of hSOD1G93A rats with doses equivalent to the concentrations that killed AbAs caused a marginally significant delay in survival, partially preserved the body weight and soleus muscle mass and improved the integrity of the neuromuscular junction. Reduced motor neuron death and glial reactivity was also found and likely related to decreased inflammation and matrix metalloproteinase-2 and -9. Thus, in spite that new experimental designs aimed at extending the lifespan of hSOD1G93A rats are needed, improvements observed upon violacein treatment suggest a significant therapeutic potential that deserves further studies.
SIRT6 stabilization and cytoplasmic localization in macrophages regulates acute and chronic inflammation in mice
J Biol Chem 2022 298(3):101711
Mariana Bresque 1 , Karina Cal 2 , Valentina Pérez-Torrado 3 , Laura Colman 4 , Jorge Rodríguez-Duarte 5 , Cecilia Vilaseca 6 , Leonardo Santos 7 , María Pía Garat 7 , Santiago Ruiz 7 , Frances Evans 8 , Rosina Dapueto 1 , Paola Contreras 9 , Aldo Calliari 2 , Carlos Escande 10
1 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay; Laboratory of Vascular Biology and Drug Development, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay. 2 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay; Departamento de Biociencias, Facultad de Veterinaria, Universidad de la República (UdelaR), Montevideo, Uruguay. 3 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay; Departamento de Inmunobiología, Facultad de Medicina, Universidad de la República (UdelaR), Montevideo, Uruguay; Área Inmunología, Departamento de Biociencias, Facultad de Química, Universidad de la República (UdelaR), Montevideo, Uruguay. 4 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay; Departamento de Química Orgánica, Facultad de Química, Universidad de la República (UdelaR), Montevideo, Uruguay. 5 Laboratory of Vascular Biology and Drug Development, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay. 6 Departamento de Fisiología, Facultad de Medicina, Universidad de la República (UdelaR), Montevideo, Uruguay. 7 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay. 8 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República (UdelaR), Montevideo, Uruguay; Laboratory of Neuroinflammation and Gene Therapy, Institut Pasteur Montevideo, Montevideo, Uruguay. 9 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay; Departamento de Fisiología, Facultad de Medicina, Universidad de la República (UdelaR), Montevideo, Uruguay. 10 Laboratory of Metabolic Diseases and Aging, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay. Electronic address: escande@pasteur.edu.uy.
DOI: 10.1016/j.jbc.2022.101711
PMID: 35150745
Pubmed: https://pubmed.ncbi.nlm.nih.gov/35150745
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(22)00151-X
Abstract:
Acute and chronic inflammations are key homeostatic events in health and disease. Sirtuins (SIRTs), a family of NAD-dependent protein deacylases, play a pivotal role in the regulation of these inflammatory responses. Indeed, SIRTs have anti-inflammatory effects through a myriad of signaling cascades, including histone deacetylation and gene silencing, p65/RelA deacetylation and inactivation, and nucleotide‑binding oligomerization domain, leucine rich repeat, and pyrin domain‑containing protein 3 inflammasome inhibition. Nevertheless, recent findings show that SIRTs, specifically SIRT6, are also necessary for mounting an active inflammatory response in macrophages. SIRT6 has been shown to positively regulate tumor necrosis factor alpha (TNFα) secretion by demyristoylating pro-TNFα in the cytoplasm. However, how SIRT6, a nuclear chromatin-binding protein, fulfills this function in the cytoplasm is currently unknown. Herein, we show by Western blot and immunofluorescence that in macrophages and fibroblasts there is a subpopulation of SIRT6 that is highly unstable and quickly degraded via the proteasome. Upon lipopolysaccharide stimulation in Raw 264.7, bone marrow, and peritoneal macrophages, this population of SIRT6 is rapidly stabilized and localizes in the cytoplasm, specifically in the vicinity of the endoplasmic reticulum, promoting TNFα secretion. Furthermore, we also found that acute SIRT6 inhibition dampens TNFα secretion both in vitro and in vivo, decreasing lipopolysaccharide-induced septic shock. Finally, we tested SIRT6 relevance in systemic inflammation using an obesity-induced chronic inflammatory in vivo model, where TNFα plays a key role, and we show that short-term genetic deletion of SIRT6 in macrophages of obese mice ameliorated systemic inflammation and hyperglycemia, suggesting that SIRT6 plays an active role in inflammation-mediated glucose intolerance during obesity.