Valery Grinevich Group
Summary of Research
There are three directions, which we are following in our research: 1) Employment and generation of transgenic mice to dissect the role of various genes in the regulation of stress responses; 2) Exploration of functional connectivity of hypothalamic neurons by viral techniques; 3) Application of immediate early gene-based indicator mice for monitoring activity of affected by stress brain circuitries.
To manipulate the hypothalamic-pituitary-adrenal (HPA) axis we are in the process of generating and analyzing transgenic BAC mouse lines, expressing Cre recombinase and GFP under the control of the promoter of corticotropin-releasing hormone (CRH) gene.
To trace anatomical connections of hypothalamic neurons we apply various recombinant viruses such as lentivirus, adeno-associated virus, and pseudotyped rabies virus. The cell-type specific targeting of hypothalamic neurons by virus is successfully achieved by using viral constructs containing evolutionarily-conserved promoter sequences of genes of hypothalamic neurohormones. In addition, we are combining viral techniques with transgenic Cre-expressing mouse lines in order to express genes of interest upon Cre recombinase activity. Both these strategies will lead to cell-specific expression of anterograde and retrograde markers in certain types of hypothalamic neurons, such as oxytocin, vasopressin and CRH neurons.
To monitor the activity of affected by stress brain circuits we apply our recently generated BAC indicator mouse, which expresses 4-hour half-life green fluorescent protein (d4eGFP) under the control of a promoter of activity regulated cytoskeletal protein (Arc). The use of this and other immediate early gene-based indicator mice will allow us to monitor activity of impaired brain circuits in animal models of stress-related diseases, as well as to screen pharmacological compounds for their ability to normalize activity in specific brain regions.
