Our lab aims to combine mouse-human behavioural and genetics data, taking advantage of cutting-edge techniques available in rodents to dissect in vivo circuit-, time- and cell-specific mechanisms of cognitive and social processes. Our behavioural, genetics, pharmacological and molecular expertise is integrated with the interdisciplinary environment of the Istituto Italiano di Tecnologia, with state of the art equipment and recognized scientists with diverse backgrounds (e.g. neuroimaging, electrophysiology, molecular biology, nanotechnology, bioinformatics, drug discovery, computation, robotics, smart materials etc.).
Below are some of the main technologies we incorporate into our research to answer our scientific questions:
Biobanks & Patient Samples
We directly test and translate our preclinical findings into the clinics. Thanks to our collaborations with different clinical centers in Italy and around the world, we collect data on human neurocognitive and clinical assessments as well as biological samples (brain and blood samples) that are processed and analyzed in our laboratory. We also have exclusive access to different human databases.
To study gene functions and diseases mechanisms in vivo, we use genetically modified mice, as examples:
- Mouse lines with clinically-relevant genetic variants (e.g. 22q11.2, 16p11.2, COMT Val, COMT ko, Dysbindin-1, 1A, 1C, Arc/Arg3.1, D2L, D3, DAT, CRF/CRF1, CRF/CRF2, NRG1-IV, CB1, OXT, and double mutants with different combinations).
- Floxed and Cre transgenic mouse lines to investigate site-, time- and cell-type specific mechanisms.
Main brain regions of interest: prefrontal cortex, amygdala, hypothalamus, postero-parietal cortex.
Developmental time: perinatal, pre-adolescence, adolescence, adulthood.
Cell types: pyramidal neurons, SOM+ and PV+ interneurons, astrocytes, and microglia.
Systems: dopamine, oxytocin, CRF and cannabinoids.
Measuring simple and complex mouse behaviour
Visualizing the brain
We perform ex vivo and in vivo imaging analyses of the mouse brain to investigate the circuits of social/cognitive brain. We also study the morphological alterations in neurons and glial cell induced by genetic variations and/or drug administrations at different stages of development.
In vivo miniscopes
Miniscopes enables one-photon epifluorescent imaging of calcium dynamics in freely moving mice. With this technique, we map the brain activity with single-cell resolution, enabling us to compare the physiological activity of a specific cell type (e.g. neurons or astrocytes) during different social cognitive tasks within different conditions and between different days.
In vivo Fiber Photometry
Fibre photometry uses the intensity-based genetically encoded indicators for calcium (GCaMP) or dopamine (dLight1) to assess the neural activity of defined subpopulations of neurons through an optic fibre in freely moving animals. This technique allows us to record with a high spatiotemporal resolution neural activity in the nucleus accumbens, amygdala and prefrontal cortex, while animals are performing our cognitive or social tasks.
In vivo Electrophysiology
To study the neural substrates of cognitive and social behaviours we employ tetrode and silicon probe recordings in freely moving mice while performing social- and higher-order cognitive tasks. These methods allow us to analyze the activity of single cells as well as the activity of neuronal networks on a millisecond time scale. To address the contribution of defined cell populations to the neuronal network activity we also combine electrophysiological recordings with optogenetic manipulations.
In vivo microdialysis
Intracerebral in vivo microdialysis allows the collection of small-molecular-weight substances from brain interstitial space. Here we use this technique coupled to high-pressure liquid chromatography to monitor in a time-dependent manner the outflow of neurotransmitters (dopamine, norepinephrine, serotonin, GABA, glutamate) and their metabolites in discrete brain regions, such as the nucleus accumbens and the prefrontal cortex.
In vivo Optogenetics and Chemogenetics
To gain new mechanistic insights, we use these techniques to activate or inhibit with temporal and spatial specificity targeted subcellular populations or specific neuronal pathways in freely moving mice while performing our cognitive and social tasks.
Cell and molecular biology
Despite we are prevalently an in vivo laboratory, we employ classical techniques to both validate our models and get insights into the cellular and molecular mechanisms involved in the behavioural alterations we investigate.
We perform mRNA and protein quantifications through real-time PCR, western blot and ELISA tests. We analyse cell populations proportions in neural and immune samples, by fluorescence-based flow cytometry, and select specific cell populations by fluorescence-activated cell sorting (FACS) for gene expression analyses. Finally, we investigate cell and tissue morphology by cutting-edge fluorescence and confocal microscopes.