Mouse genetics and target-specific brain manipulation
To study gene function and disease mechanisms in-vivo, we use transgenic mouse models, together with site-specific manipulations of neuronal targets using viral injections. These tools include:
- Classical loss- and gain-of-function mouse lines: Knock-out and transgenic mice, injection of gene-silencing or over-expression vectors
- Tools to control target expression in a cell-type and time-specific manner: Cell-specific, drug- or activity-inducible Cre and Flp recombinases
- Cell-specific enrichment technologies: RiboTag mice, tAGO2 mice, INTACT mice, RNA-guided nucleases – e.g. CRISPR/Cas system
- Chemogenetic and optogenetic approaches for neuronal activation or inhibition in a cell- and site-specific manner
Visualizing the brain
To investigate neuronal circuits and visualize expression of our target genes and proteins, we use advanced tissue staining and fluorescent microscopy techniques, including single-molecule RNA hybridization, and confocal and light-sheet imaging. For non-invasive assessment of structural, functional and metabolic markers in living animals, we use magnetic resonance imaging techniques including structural MRI and manganese-enhanced MRI (MEMRI) for structural or functional characterization.
Measuring simple & complex mouse behavior
We use classical, standardized assays to measure mouse behavior following manipulation: Anxiety and depression like behavior are assessed by the open field, elevated plus maze, dark-light box, and sucrose preference tests. Cognitive assays include object recognition, fear conditioning, the Morris Water maze, and five-choice serial-reaction time task. To study a more complex and naturalistic behavioral repertoire, we developed a novel system called the Social Box for tracking multiple animals in an enriched environment, with minimal intervention and across long time spans. This allows us to characterize the nature of free behavior and interactions in a group setting.
Regulation of gene expression
We study the regulation of stress-related gene expression (including alternative splicing and polyadenylation) using both transcriptome-wide measurements, such as RNA-sequencing, and candidate-driven approaches like real-time PCR and in-situ hybridization. In addition to mRNA, our lab also focuses on measuring microRNA expression – using real-time PCR and dedicated small RNA-Sequencing and in-situ hybridization protocols – and to investigate their regulatory functions, using in vitro studies, genetic mouse models and virus-based manipulations. We also investigate how epigenetic mechanisms like DNA methylation regulate gene expression during stress, using DNA-pyrosequencing, traditional BS-sequencing, TAB-sequencing and genetic mouse models.
We study the neural substrates of behavior using patch-clamp and field potential recordings in-vitro (cell culture, organotypic and acute brain slices), as well as incorporating state-of-the-art optogenetic and chemogenetic manipulations.
Telemetry, Sleep and Physiology
Using cage-based and in vivo telemetry systems, we investigate physiological parameters such as heart rate, locomotor activity, respiration, and metabolic activity, as well as brain activity (ECG, EEG) in awake and sleeping animals. We are also developing methods to incorporate such telemetric systems into the enriched group environment of the Social Box.
Biobanks & Patient Samples
In addition to mouse models, we also study stress-related neurobiological functions directly in humans. We collect biological samples from study participants the clinic at Schneider Hospital and process them to obtain measures of DNA, RNA, miRNA, and proteomics. This clinic at Schneider Hospital is responsible for diagnosis, assessment and treatment of children and adolescents, aged 6-18, suffering from mental disorders. All clinical evaluations and assessments, as well as biological sample collection, are conducted by the team at the clinic, while molecular assessments and analyses are conducted at the lab in the Weizmann Institute.