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Publicaciones del Departamento de Histología y Embriología
Phasor-based multi-harmonic unmixing for in-vivo hyperspectral imaging
Methods Appl Fluoresc 2022 11(1)
Alexander Vallmitjana 1 , Paola Lepanto 2 , Florencia Irigoin 2 3 4 , Leonel Malacrida 4 5
1 Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, United States of America. 2 Human Molecular Genetics Lab, Institut Pasteur de Montevideo, Montevideo, Mataojo 2020, CP11400, Uruguay. 3 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 4 Advanced Bioimaging Unit, Institut Pasteur of Montevideo and Universidad de la República, Montevideo, Uruguay. 5 Departamento de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
DOI: 10.1088/2050-6120/ac9ae9
PMID: 36252561
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36252561
Texto completo: https://doi.org/10.1088/2050-6120/ac9ae9
Abstract:
Hyperspectral imaging (HSI) is a paramount technique in biomedical science, however, unmixing and quantification of each spectral component is a challenging task. Traditional unmixing relies on algorithms that need spectroscopic parameters from the fluorescent species in the sample. The phasor-based multi-harmonic unmixing method requires only the empirical measurement of the pure species to compute the pixel-wise photon fraction of every spectral component. Using simulations, we demonstrate the feasibility of the approach for up to 5 components and explore the use of adding a 6th unknown component representing autofluorescence. The simulations show that the method can be successfully used in typical confocal imaging experiments (with pixel photon counts between 101and 103). As a proof of concept, we tested the method in living cells, using 5 common commercial dyes for organelle labeling and we easily and accurately separate them. Finally, we challenged the method by introducing a solvatochromic probe, 6-Dodecanoyl-N,N-dimethyl-2-naphthylamine (LAURDAN), intended to measure membrane dynamics on specific subcellular membrane-bound organelles by taking advantage of the linear combination between the organelle probes and LAURDAN. We succeeded in monitoring the membrane order in the Golgi apparatus, Mitochondria, and plasma membrane in the samein-vivocell and quantitatively comparing them. The phasor-based multi-harmonic unmixing method can help expand the outreach of HSI and democratize its use by the community for it does not require specialized knowledge.
Standards in semen examination: publishing reproducible and reliable data based on high-quality methodology
Hum Reprod 2022 37(11):2497-2502
Lars Björndahl 1 , Christopher L R Barratt 2 , David Mortimer 3 , Ashok Agarwal 4 , Robert J Aitken 5 , Juan G Alvarez 6 7 , Natalie Aneck-Hahn 8 , Stefan Arver 1 , Elisabetta Baldi 9 , Lluís Bassas 10 , Florence Boitrelle 11 12 , Riana Bornman 13 , Douglas T Carrell 14 15 , José A Castilla 16 17 , Gerardo Cerezo Parra 18 , Jerome H Check 19 20 , Patricia S Cuasnicu 21 , Sally Perreault Darney 22 23 , Christiaan de Jager 24 , Christopher J De Jonge 25 , Joël R Drevet 26 , Erma Z Drobnis 27 , Stefan S Du Plessis 28 , Michael L Eisenberg 29 30 , Sandro C Esteves 31 32 33 , Evangelini A Evgeni 34 35 , Alberto Ferlin 36 , Nicolas Garrido 37 , Aleksander Giwercman 38 , Ilse G F Goovaerts 39 , Trine B Haugen 40 , Ralf Henkel 41 42 , Lars Henningsohn 43 44 , Marie-Claude Hofmann 45 , James M Hotaling 46 , Piotr Jedrzejczak 47 , Pierre Jouannet 48 , Niels Jørgensen 49 50 , Jackson C Kirkman Brown 51 52 53 , Csilla Krausz 54 , Maciej Kurpisz 55 , Ulrik Kvist 1 , Dolores J Lamb 56 , Hagai Levine 57 , Kate L Loveland 58 , Robert I McLachlan 59 , Ali Mahran 60 61 , Liana Maree 42 , Sarah Martins da Silva 2 , Michael T Mbizvo 62 , Andreas Meinhardt 63 , Roelof Menkveld 64 , Sharon T Mortimer 3 65 , Sergey Moskovtsev 66 67 , Charles H Muller 68 , Maria José Munuce 69 , Monica Muratori 54 , Craig Niederberger 70 71 , Cristian O'Flaherty 72 , Rafael Oliva 73 74 , Willem Ombelet 75 76 , Allan A Pacey 77 78 , Michael A Palladino 79 , Ranjith Ramasamy 80 , Liliana Ramos 81 , Nathalie Rives 82 , Eduardo Rs Roldan 83 , Susan Rothmann 84 , Denny Sakkas 85 , Andrea Salonia 86 87 , Maria Cristina Sánchez-Pozo 88 , Rosanna Sapiro 89 , Stefan Schlatt 90 , Peter N Schlegel 91 , Hans-Christian Schuppe 92 , Rupin Shah 93 , Niels E Skakkebæk 49 , Katja Teerds 94 , Igor Toskin 95 , Herman Tournaye 96 , Paul J Turek 97 , Gerhard van der Horst 98 99 100 , Monica Vazquez-Levin 101 , Christina Wang 102 103 , Alex Wetzels 104 , Theodosia Zeginiadou 105 106 , Armand Zini 107
1 ANOVA, Karolinska University Hospital and Karolinska Institutet, Stockholm, Sweden. 2 Reproductive Medicine Research Group, Division of Systems Medicine, School of Medicine, Ninewells Hospital and Medical School, University of Dundee, Dundee, UK. 3 Oozoa Biomedical Inc., West Vancouver, BC, Canada. 4 Case Western Reserve University, Moreland Hills, OH, USA. 5 Priority Research Centre for Reproductive Science, Faculty of Science and Faculty of Health & Medicine, University of Newcastle, Callaghan, NSW, Australia. 6 Centro Androgen, La Coruña, Spain. 7 Harvard Medical School, Boston, MA, USA. 8 Department of Urology, University of Pretoria, Pretoria, South Africa. 9 Department of Experimental and Clinical Medicine, University of Florence, Florence, Tuscany, Italia. 10 Andrology Department, Laboratory of Andrology and Sperm Bank, Fundació Puigvert, Barcelona, Spain. 11 Department of Reproductive Biology, Fertility Preservation, Andrology, CECOS, Poissy Hospital, Poissy, France. 12 Paris Saclay University, UVSQ, INRAE, BREED, Jouy-en-Josas, France. 13 School of Health Systems and Public Health, University of Pretoria, Pretoria, South Africa. 14 Andrology and IVF Laboratory, Division of Urology, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA. 15 Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT, USA. 16 GAMETIA Biobank, Granada, Spain. 17 Hospital Universitario Virgen de las Nieves and Instituto de Investigación Biosanitaria ibs. GRANADA, Granada, Spain. 18 LAFER Sperm Bank, Tuxpan 10-606, Roma Sur, C.P. 06760, Cuauhtémoc, Mexico City, Mexico. 19 Robert Wood Johnson Medical School at Camden, The University of Medicine and Dentistry of New Jersey, Camden, NJ, USA. 20 Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology & Infertility, Cooper Hospital/University Medical Center, Melrose Park, PA, USA. 21 Instituto de Biología y Medicina Experimental (IbyME-CONICET), Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina. 22 US EPA, Cary, NC, USA. 23 US NIH, Cary, NC, USA. 24 Faculty of Health Science, University of Pretoria, Pretoria, South Africa. 25 University of Minnesota Medical Center, University of Minnesota, Minneapolis, MN, USA. 26 Université Clermont Auvergne/CNRS/INSERM-GreD Institute, Clermont-Ferrand, France. 27 School of Medicine, University of Missouri, Columbia, MI, USA. 28 College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates. 29 Male Reproductive Medicine and Surgery, Stanford University School of Medicine, Stanford, CA, USA. 30 Department of Urology, Stanford University School of Medicine, Stanford, CA, USA. 31 ANDROFERT, Andrology and Human Reproduction Clinic, Campinas, Brazil. 32 Department of Surgery (Division of Urology), University of Campinas (UNICAMP), Campinas, Brazil. 33 Faculty of Health, Aarhus University, Aarhus C, Denmark. 34 CRYOGONIA Cryopreservation Bank, Athens, Greece. 35 Laboratory of Physiology, Department of Medicine, Democritus University of Thrace, Greece. 36 Unit of Andrology and Reproductive Medicine, Department of Medicine, University of Padova, Padova, Italia. 37 IVI Foundation, Health Research Institute La Fe, Valencia, Spain. 38 Department of Translational Medicine, Lund University, Malmö, Sweden. 39 Antwerp University Hospital, Edegem, Belgium. 40 Department of Life Sciences and Health, Oslo Metropolitan University, Oslo, Norway. 41 Department of Metabolism, Digestion & Reproduction, Imperial College London, London, UK. 42 Department of Medical Bioscience, University of the Western Cape, Bellville, South Africa. 43 Division of Urology, Department of CLINTEC, Karolinska Institutet, Stockholm, Sweden. 44 Department of Urology, Karolinska University Hospital, Stockholm, Sweden. 45 Department of Endocrine Neoplasia & Hormonal Disorders, University of Texas MD Anderson Cancer Center, Houston, TX, USA. 46 Division of Urology, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA. 47 Department of Cell Biology, Poznan University of Medical Science, Poznan, Poland. 48 Université Paris Descartes, Paris, France. 49 Department of Growth and Reproduction, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark. 50 International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC), Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark. 51 Centre for Human Reproductive Science (ChRS), UK. 52 College of Medical & Dental Sciences, University of Birmingham, UK. 53 Birmingham Women's and Children's NHS Foundation Trust, UK. 54 Department of Biomedical, Experimental and Clinical Sciences "Mario Serio", University of Florence, Florence, Italy. 55 Department of Reproductive Biology and Stem Cells, Institutet of Human Genetics, Poznan, Poland. 56 Brady Department of Urology, Center for Reproductive Genomics and Englander Institute for Precision Medicine, Weill Cornell Medical College, Cornell University, New York, NY, USA. 57 Braun School of Public Health and Community Medicine, Hadassah Medical Center, The Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel. 58 Hudson Institute, Centre for Reproductive Health, Monash University, Clayton, VIC, Australia. 59 Hudson Institute of Medical Research, Centre for Endocrinology and Metabolism, Monash University, Clayton, VIC, Australia. 60 Dermatology and Andrology Department, Assiut University Hospital, Assiut, Egypt. 61 Faculty of Medicine, Assiut University, Assiut, Egypt. 62 Country Director, Population Council (International Programs). 63 Department of Anatomy and Cell Biology, Justus-Liebig-University of Giessen, Giessen, Germany. 64 Department of Obstetrics and Gynaecology, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg, South Africa. 65 Division of REI, Department of Obstetrics & Gynaecology, Faculty of Medicine, University of British Columbia, Vancouver, BC, Canada. 66 Department of Laboratory Medicine & Pathobiology, University of Toronto, Toronto, ON, Canada. 67 CreATe Fertility Centre, Toronto, ON, Canada. 68 Male Fertility Laboratory, Department of Urology, University of Washington School of Medicine, Seattle, WA, USA. 69 Laboratorio de Medicina Reproductiva, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina. 70 Department of Urology, UIC College of Medicine, IL, USA. 71 Department of Bioengineering, UIC College of Engineering, IL, USA. 72 Department of Surgery (Urology Division), McGill University, Montréal, QC, Canada. 73 Molecular Biology of Reproduction and Development Group, Biomedical Research Institute August Pi I Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain. 74 Hospital Clínic, University of Barcelona, Barcelona, Spain. 75 Genk Institute for Fertility Technology, Genk, Belgium. 76 Department of Obstetrics and Gynaecology, ZOL Hospitals and Hasselt University, Genk, Belgium. 77 Department of Oncology and Metabolism, University of Sheffield, Sheffield, UK. 78 Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, UK. 79 Bloomfield College, Bloomfield, NJ, USA. 80 Department of Urology, Miller School of Medicine, University of Miami, Miami, FL, USA. 81 Division of Reproductive Medicine, Department of Obstetrics and Gynaecologie, Radboud UMC, Nijmegen, The Netherlands. 82 Service Laboratoire de Biologie de la Reproduction-CECOS, Equipe Physiopathologie Surrénalienne et Gonadique, Unité Inserm 1239 NorDic, CHU-Hôpitaux de Rouen, UFR Santé-Université de Rouen, Rouen, France. 83 Department of Biodiversity and Evolutionary Biology, Museo Nacional de Ciencias Naturales (CSIC), Madrid, Spain. 84 Fertility Solutions Inc., Warrensville Heights, OH, USA. 85 Boston IVF, Boston, MA, USA. 86 University Vita-Salute San Raffaele, Milan, Italy. 87 Division of Experimental Oncology/Unit of Urology, URI, IRCCS Ospedale San Raffaele, Milan, Italy. 88 Department of Clinical Chemistry and Molecular Biology, Virgen del Rocío University Hospital, Seville, Spain. 89 Depto de Histologia y Embriología, Facultad de Medicina, Gral. Flores, Uruguay. 90 Centre of Reproductive Medicine and Andrology, Münster, Germany. 91 Department of Urology, Weill Cornell Medicine, New York, NY, USA. 92 Section of Andrology, Department of Urology, Pediatric Urology & Andrology, Justus-Liebig-University/University Hospital of Giessen-Marburg, Giessen, Germany. 93 Lilavati Hospital & Research Centre, Mumbai, India. 94 Human and Animal Physiology, Wageningen University, Wageningen, The Netherlands. 95 WHO Department of Sexual and Reproductive Health and Research (includes the UNDP/UNFPA/UNICEF/WHO/World Bank Special Programme of Research, Development and Research Training in Human Reproduction-HRP), Geneva, Switzerland. 96 Centre for Reproductive Medicine, Vrije Universiteit Brussel, Brussels, Belgium. 97 The Turek Clinic, San Francisco, CA, USA. 98 Medical Bioscience, University of the Western Cape, Bellville, South Africa. 99 Physiology Medical School, Stellenbosch University, Stellenbosch, South Africa. 100 Department of Animal Science, Stellenbosch University, Stellenbosch, South Africa. 101 IBYME, CONICET-FIBYME, Buenos Aires, Argentina. 102 Clinical and Translational Science Institute, The Lundquist Institute, Torrance, CA, USA. 103 Division of Endocrinology, Department of Medicine, Harbor-UCLA Medical Center, Torrance, CA, USA. 104 Fertility Laboratory, Radboud University Medical Centre, Nijmegen, The Netherlands. 105 Thessaloniki Andrology Laboratory-Hellenic Sperm Bank, Thessaloniki, Greece. 106 Laboratory of Histology-Embryology, Medical School, University of Athens, Athens, Greece. 107 Division of Urology, Department of Surgery, St Mary's Hospital, McGill University, Montreal, Canada.
DOI: 10.1093/humrep/deac189
PMID: 36112046
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36112046
Texto completo: https://academic.oup.com/humrep/article-lookup/doi/10.1093/humrep/deac189
Abstract:
Biomedical science is rapidly developing in terms of more transparency, openness and reproducibility of scientific publications. This is even more important for all studies that are based on results from basic semen examination. Recently two concordant documents have been published: the 6th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen, and the International Standard ISO 23162:2021. With these tools, we propose that authors should be instructed to follow these laboratory methods in order to publish studies in peer-reviewed journals, preferable by using a checklist as suggested in an Appendix to this article.
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.