School of Medicine and Health Sciences Poster Presentations
Localized, Ubiquitous Cell Ablation as a Novel, Non-Traumatic Model for Brain Atrophy
Poster Number
278
Document Type
Poster
Publication Date
3-2016
Abstract
The ability to selectively ablate cells in the central nervous system (CNS) based on their gene expression profile has been a useful tool in studying many forms of neurological disease. In previous studies, we have shown that we can locally or systemically ablate cells using a combination of a transgenic mouse line and a chemical inducer of dimerization (CID) to manipulate the caspase-9 mediated apoptotic pathway, thus producing cell death through activation of an inducible caspase-9 (iCP-9) signaling cascade. Our current study builds upon our previous model by making use of a site-specific transgene, the cytomegalovirus gene (CMV), as opposed to a random gene insertion, to allow for ubiquitous expression throughout the genome. By driving the CMV gene via a Cre-recombinase transgenic line and crossing it with our iCP-9 flox animal, we have engineered a transgenic mouse that globally expresses our transgene of interest on which to test the utility of chemically induced dimerization as an experimental model for controlling localized cell death. In order to experimentally test the efficacy of our system for inducing cell ablation in vitro, we prepared primary brain cell cultures from individual CMVcre; iCP9 transgenic and wild type mice at P2 and P3. Our data shows that within 24 hours of CID treatment, significant cell loss is detectable in cmvcre-iCP9 transgenic cultures, but not in wild type cultures. We have also shown, in vivo, that with direct injection of CID into the cerebellum of both adult (4 months) and developing animals (2 weeks), we can drive a localized region of complete cell death at just 2 days post injection. Therefore, our data demonstrates that the caspase-9 mediated apoptotic pathway can be manipulated using a chemical inducer of dimerization to induce localized, ubiquitous cell ablation in CMVcre; iCP-9 transgenic mice. These preliminary data suggest that our unique transgenic approach to controlling cell death may serve as a useful non-traumatic and non-immune-mediated alternative model for studying traumatic brain injury.
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Open Access
1
Localized, Ubiquitous Cell Ablation as a Novel, Non-Traumatic Model for Brain Atrophy
The ability to selectively ablate cells in the central nervous system (CNS) based on their gene expression profile has been a useful tool in studying many forms of neurological disease. In previous studies, we have shown that we can locally or systemically ablate cells using a combination of a transgenic mouse line and a chemical inducer of dimerization (CID) to manipulate the caspase-9 mediated apoptotic pathway, thus producing cell death through activation of an inducible caspase-9 (iCP-9) signaling cascade. Our current study builds upon our previous model by making use of a site-specific transgene, the cytomegalovirus gene (CMV), as opposed to a random gene insertion, to allow for ubiquitous expression throughout the genome. By driving the CMV gene via a Cre-recombinase transgenic line and crossing it with our iCP-9 flox animal, we have engineered a transgenic mouse that globally expresses our transgene of interest on which to test the utility of chemically induced dimerization as an experimental model for controlling localized cell death. In order to experimentally test the efficacy of our system for inducing cell ablation in vitro, we prepared primary brain cell cultures from individual CMVcre; iCP9 transgenic and wild type mice at P2 and P3. Our data shows that within 24 hours of CID treatment, significant cell loss is detectable in cmvcre-iCP9 transgenic cultures, but not in wild type cultures. We have also shown, in vivo, that with direct injection of CID into the cerebellum of both adult (4 months) and developing animals (2 weeks), we can drive a localized region of complete cell death at just 2 days post injection. Therefore, our data demonstrates that the caspase-9 mediated apoptotic pathway can be manipulated using a chemical inducer of dimerization to induce localized, ubiquitous cell ablation in CMVcre; iCP-9 transgenic mice. These preliminary data suggest that our unique transgenic approach to controlling cell death may serve as a useful non-traumatic and non-immune-mediated alternative model for studying traumatic brain injury.
Comments
Presented at: GW Research Days 2016