Universidad de la República

8 ARTICLES PUBLISHED IN JoVE

Biology

Simple and Efficient Technique for the Preparation of Testicular Cell Suspensions

Rosana Rodríguez-Casuriaga¹, Gustavo A. Folle², Federico Santiñaque², Beatriz López-Carro², Adriana Geisinger^1,3

¹Departamento de Biología Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), ²Servicio de Citometría de Flujo y Clasificación Celular (SECIF), Instituto de Investigaciones Biológicas Clemente Estable, ³Sección Bioquímica, Facultad de Ciencias, Universidad de la República

A novel protocol for the mechanical preparation of testicular cell suspensions from rodent material, avoiding enzymes and detergents, is described. The method is very simple, fast, reproducible, and renders good quality cell suspensions, which are suitable for flow sorting and RNA extraction.

JoVE Core

Sublingual Immunotherapy as an Alternative to Induce Protection Against Acute Respiratory Infections

Natalia Muñoz-Wolf^1,2, Analía Rial¹, José M. Saavedra¹, José A. Chabalgoity¹

¹Departamento de Desarrollo Biotecnológico, Universidad de la República, ²Present Affiliation: Trinity Biomedical Science Institute, Trinity College Dublin

The present work illustrates the convenience of using sublingual immunotherapy to boost the innate immune response in the lungs and confer protection against acute pneumococcal pneumonia in mouse.

Developmental Biology

Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos

Daniel Prieto¹, Gonzalo Aparicio^1,2, Matías Machado¹, Flavio R. Zolessi^1,2

¹Institut Pasteur de Montevideo, ²Sección Biología Celular, Facultad de Ciencias, Universidad de la República

A method for fluorescent staining of fixed biological material with the specific DNA label methyl green is described. Methyl green is used in a diluted aqueous solution and is very resistant to photobleaching. Its far-red emission allows for deep specimen imaging, making it particularly adequate for whole embryos.

Developmental Biology

A Rapid and Efficient Method to Dissect Pupal Wings of Drosophila Suitable for Immunodetections or PCR Assays

Carmen Bolatto¹, Cristina Parada¹, Victoria Colmenares¹

¹Departamento de Histología y Embriología, Facultad de Medicina, Laboratorio de Biología del Desarrollo

The ability to accurately detect transcripts or proteins in Drosophila tissues is critical for studying their abundance and localization related to the development process. Here is the description of a straightforward procedure to dissect pupal wings. These wings can be used as samples in several techniques (immunohistochemistry, PCR assay, etc.).

Neuroscience

Dissection of Single Skeletal Muscle Fibers for Immunofluorescent and Morphometric Analyses of Whole-Mount Neuromuscular Junctions

Carmen Bolatto^*1, Silvia Olivera-Bravo^*2, Sofía Cerri^1,2

¹Developmental Biology Laboratory, Histology and Embryology Department, Faculty of Medicine, Universidad de la República, ²Cell and Molecular Neurobiology Laboratory, Clemente Estable Biology Research Institute (IIBCE), Ministerio de Educación y Cultura

The ability to accurately detect neuromuscular junction components is crucial in evaluating modifications in its architecture because of pathological or developmental processes. Here we present a complete description of a straightforward method to obtain high-quality images of whole-mount neuromuscular junctions that can be used to perform quantitative measurements.

Developmental Biology

Atomic Force Microscopy to Study the Physical Properties of Epidermal Cells of Live Arabidopsis Roots

Ines Rauschert^*2, Juan Claudio Benech^*2, Martha Sainz¹, Omar Borsani¹, Mariana Sotelo-Silveira¹

¹Laboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República (UdelaR), ²Laboratorio de Señalización Celular y Nanobiología, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE)

The atomic force microscopy indentation protocol offers the possibility to dissect the role of the physical properties of the cell wall of a particular cell of a tissue or organ during normal or constrained growth (i.e., under water deficit).

Developmental Biology

Polysome Purification from Soybean Symbiotic Nodules

María Martha Sainz¹, Carla Valeria Filippi¹, Guillermo Eastman^2,3, Mariana Sotelo-Silveira¹, C. Mauro Martinez¹, Omar Borsani¹, José Sotelo-Silveira^2,4

¹Laboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, ²Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, ³Department of Biology, University of Virginia, ⁴Departamento de Biología Celular y Molecular, Facultad de Ciencias, Universidad de la República

This protocol describes a method for eukaryotic polysome purification from intact soybean nodules. After sequencing, standard pipelines for gene expression analysis can be used to identify differentially expressed genes at the transcriptome and translatome levels.

Medicine

High-Resolution Respirometry to Assess Mitochondrial Function in Human Spermatozoa

Pilar Irigoyen¹, Rossana Sapiro^1,3, Adriana Cassina^2,3

¹Departamento de Histología y Embriología, Unidad Académica Histología y Embriología, ²Departamento de Bioquímica, Unidad Académica Bioquímica, ³Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República

The analysis of sperm mitochondrial function by high-resolution respirometry permits the measurement of the oxygen consumption of freely moving spermatozoa in a closed-chamber system. The technique can be applied to measure respiration in human spermatozoa, which provides information on sperm mitochondrial characteristics and integrity.