Expertise

The expertise of the Institute of Cell Biology and Neurobiology includes a wide range of methodologies and know-how:

in the field of cell biology and organisms

- Culture of in vitro stabilized tumor cell lines , preparation and maintenance of dissociated primary cultures (neuronal, glia, muscle, liver) and organotypic cultures (from central nervous system, epithelium, muscle)

- Transformation of prokaryotic cells

- Transfection and infection (retrovirus, lentivirus, adenovirus) of eukaryotic cells (stable and/or transient)

- Expression of fluorescent chimeric proteins

- Enzymatic assays in cultured cells

- Signal transduction analysis

- Assays for proliferation, migration and cell adhesion

- Analysis of programmed cell death (FACS: sub-G1 fraction, Annexin V assay, Tunel assay, mithocondrial membrane potential, ROS; WB and gene expression of main cell death associated molecules: bcl-2 family, p53, IAP proteins, caspases)

- Analysis of cellular DNA content and cell cycle (FACS: cytometric ploidy, PI staining, BrDU labeling; cell cycle checkpoint regulators)

- Analysis of RNA metabolism

- Growth, genetic manipulation and analysis of yeast

- Isolation and characterization of stem cells

-Maintenance and expansion of Schistosoma mansoni

- Flow cytometry analysis (FACS CANTO II and FACScalibur)

- Receptor binding studies and evaluation of hormone levels

in the field of molecular biology

- Extraction, purification and analysis of RNA and DNA

- "in situ" hybridization

- Southern and Northern blots

- Assays of nuclear "run off"

- DNase footprinting and EMSA

- PCR, q-RT-PCR, differential display PCR

- Analysis of gene regulatory activities (assays with reporter enzymes)

- Standard recombinant DNA techniques

- Gene cloning techniques

- RNA interference

- RNase protection assays

- Site-specific mutations of nucleic acids

- DNA microarrays and arrays PCR for the expression of genes and miRNAs in cells and tissues.

- Extracellular microRNA from cell culture and blood plasma

- Purification, isolation and characterization of extracellular vesicles (exosomes)

- Chromatin immunoprecipitation

- Sequencing and analysis of nucleic acid fragments

- Construction of retroviral vectors, lentiviral and adenoviral, production of recombinant viral particles

in the field of biochemistry

- Electrophoresis of DNA and proteins

- Protein cross-linking

- Enzyme assays

- Protein phosphorylation techniques

- Expression, purification and characterization of recombinant proteins

- Purification and analysis of proteins

- Cellular fractionation techniques

- Affinity radioligand assays for receptors and ion channels

- Immunoprecipitation and analysis of protein and protein-RNA complexes

- Determination of radical species  

in the field of immunology

- ELISA

- Preparation, purification and characterization of monoclonal and polyclonal antibodies

- Immunopurification of proteins

in the context of morphology

- Determinations and morphometric analysis

- Immunohistochemistry and immunocytochemistry techniques

(Immunofluorescence, immuno-peroxidase and electron immunomicroscopy)

- Optical microscopy, fluorescence, confocal, and videoimaging (time lapse)

- Preparation and analysis of serial sections of organs and tissues (microtome, cryostat ...)

- Visualization techniques and image processing

- High resolution micro-CT imaging, 2D / 3D quantitative analysis

as part of the analysis of behaviour and cognition in mouse and rat models

Tests for the study of:

- Motor and sensory-motor abilities (SHIRPA, developmental milestones), muscle strength (grip strength)

- Exploratory behaviour and locomotor activity (open field, running wheels, treadmill)

- Coordination and motor learning (rotarod, pole test, balance beam, foot print test)

- Episodic memory and spatial navigation (Morris water maze, radial arm maze, fear conditioning. Object displacement and object recognition, Y-maze)

- Operant conditioning (active avoidance, alternation task)

- Sensory-motor gating (acoustic startle and Pre-Pulse Inhibition)

- Depression and anxiety (elevated plus maze, light-dark, forced swim, open arena)

- Substance abuse (Conditioned Place Preference, Conditioned Taste Aversion)

- Analysis and microstructure of eating behaviour

- Perception of pain and noxious stimuli (tail flick, hot plate, von Frey, Dynamic aesthesiometer, Incapacitance, Plantar, inflammatory / postoperative pain)

- Maternal behaviour

- Physiopathology of Development

- Social behaviour

- Ultrasonic vocalizations

- Olfactory tests

in the field of pharmacology and neuropharmacology in vivo

- Behavioural pharmacology

- Screening, characterization and search for new bioactive compounds

- Stereotaxic techniques (focal lesions, injections "in situ" and reversible inactivation)

in the context of in vivo metabolism

- Analysis of the circadian caloric consumption in different nutritional conditions; interface with motor activity; Basal and total metabolism; nutritional substrate (e.g. oxidation macronutrients).

Generation of mutant mouse strains

- Production and cryopreservation of mouse embryonic stem cells (ES)

- Planning and development of targeting constructs for the generation of mouse mutants

- Extraction and collection of cultured mouse embryos (popping and flushing)

- Injection of ES cells into mouse blastocysts

- Implant of mouse embryos into surrogate mothers

- Generation of transgenic mouse models by pronuclear injection.

- Generation of transgenic mouse models using CRISPR/Cas9 technology.

- Maintenance and management of murine colonies such as knock-out, knock-in and conditional mutant models

- Xenografts in immunodeficient mice

- Pharmacological treatment in mouse models

Archiving, cryopreservation, health monitoring and phenotyping of mouse mutants strains

- EU Framework Programmes-supported, strain models submission/cryopreservation/specific-pathogen-free derivation/breeding/sanitary analysis and world-wide distribution, according to EMMA-INFRAFRONTIER validated, FELASA standard-compliant procedures (https://www.infrafrontier.eu/search?keyword=browse_strain_types)

- EU Framework Programmes-supported, strain model production, according to 
EUCOMM standardized procedures


(https://www.infrafrontier.eu/search?keyword=eucomm


- professional services for standardized, primary phenotypic analysis of mutant strain models, 
according to IMPC validated, standardized procedures 
("http://www.mousephenotype.org/impress") (http://www.mousephenotype.org/impress)