The Animal Core will provide essential services and facilities to the preclinical projects in a standardized and efficient manner, utilizing shared facilities, equipment, and personnel in a coordinated fashion. The services to be provided to each preclinical project are divided into three specific aims. Dr. Thomas Martin will oversee the Animal Core and will meet with monthly or as needed with Drs. Hayashida, Peters, and Chen to assure that the Core efficiently performs these three key functions. Dr. Martin will provide oversight for regulatory affairs within the Program regarding animal use including approval and compliance with protocols for the IACUC and Biohazard Safety Committee.
Specific Aim 1 is to provide standardized in vivo procedures in animals utilized by preclinical projects. Both projects involving animals require animal procurement and identification, surgical preparation, behavioral manipulation using the repeated social defeat (RSD) paradigm, and in vivo electrophysiological recording of neuronal activity using state-of-the-art wireless technology.
Specific Aim 2 it to provide standardized methods for immunohistochemistry across preclinical projects for verification of viral mediated protein expression in discreet populations of neurons in brain or spinal cord tissue. These services include sacrifice and transcardial perfusion with paraformaldehyde, tissue harvest, cryoprotection and cryosectioning, immunohistochemistry, and fluorescent microscopy including data collection and analysis with assistance from the Bioinformatics Core.
Specific Aim 3 is to provide novel viral constructs to be utilized by preclinical projects. The two preclinical projects require generation of novel viral constructs that will provide the necessary tools for completion of specific experiments proposed. While the individual components are available through commercial sources (Addgene), these individual components must be assembled into specific vectors for the proposed studies. Dr. Rong Chen has designed subcloning strategies for each vector. She will also coordinate confirmation of sequences through outside vendors and redesign subcloning strategies if necessary. These essential functions will standardize and coordinate services and procedures throughout the Program and provide an efficient and cost-effective structure for collaboration.
The Animal Core will provide essential services and facilities to the preclinical projects in a standardized and efficient manner, utilizing shared facilities, equipment, and personnel in a coordinated fashion. The services to be provided to each preclinical project include animal procurement, identification, and surgical preparation; behavioral manipulation and assessment; in vivo electrophysiology; immunohistochemistry; and viral vector construction.
