Kyoung-Ha So1,2, Jai Ho Choi3, Jaisan Islam4, Elina Kc4, Hyeong Cheol Moon4,5, So Yoon Won6, Hyong Kyu Kim6, Soochong Kim7, Sang-Hwan Hyun1,2, Young Seok Park1,4,5. 1. Institute for Stem Cell & Regenerative Medicine (ISCRM), Veterinary Medical Center and College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea. 2. Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Veterinary Medical Center and College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea. 3. Department of Neurosurgery, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea. 4. Department of Medical Neuroscience, College of Medicine, Chungbuk National University, Cheongju, Korea. 5. Department of Neurosurgery, Chungbuk National University Hospital, Cheongju, Korea. 6. Department of Biochemistry and Medical Research Center, Chungbuk National University, Cheongju, Korea. 7. Laboratory of Veterinary Pathology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea.
Abstract
Objective: No optimum genetic rat Huntington model both neuropathological using an adeno-associated virus (AAV-2) vector vector has been reported to date. We investigated whether direct infection of an AAV2 encoding a fragment of mutant huntingtin (AV2-82Q) into the rat striatum was useful for optimizing the Huntington rat model. Methods: We prepared ten unilateral models by injecting AAV2-82Q into the right striatum, as well as ten bilateral models. In each group, five rats were assigned to either the 2×1012 genome copies (GC)/mL of AAV2-82Q (×1, low dose) or 2×1013 GC/mL of AAV2-82Q (×10, high dose) injection model. Ten unilateral and ten bilateral models injected with AAV-empty were also prepared as control groups. We performed cylinder and stepping tests 2, 4, 6, and 8 weeks after injection, tested EM48 positive mutant huntingtin aggregates. Results: The high dose of unilateral and bilateral AAV2-82Q model showed a greater decrease in performance on the stepping and cylinder tests. We also observed more prominent EM48-positive mutant huntingtin aggregates in the medium spiny neurons of the high dose of AAV2-82Q injected group. Conclusion: Based on the results from the present study, high dose of AAV2-82Q is the optimum titer for establishing a Huntington rat model. Delivery of high dose of human AAV2-82Q resulted in the manifestation of Huntington behaviors and optimum expression of the huntingtin protein in vivo.
Objective: No optimum genetic ratHuntington model both neuropathological using an adeno-associated virus (AAV-2) vector vector has been reported to date. We investigated whether direct infection of an AAV2 encoding a fragment of mutant huntingtin (AV2-82Q) into the rat striatum was useful for optimizing the Huntingtonrat model. Methods: We prepared ten unilateral models by injecting AAV2-82Q into the right striatum, as well as ten bilateral models. In each group, five rats were assigned to either the 2×1012 genome copies (GC)/mL of AAV2-82Q (×1, low dose) or 2×1013 GC/mL of AAV2-82Q (×10, high dose) injection model. Ten unilateral and ten bilateral models injected with AAV-empty were also prepared as control groups. We performed cylinder and stepping tests 2, 4, 6, and 8 weeks after injection, tested EM48 positive mutant huntingtin aggregates. Results: The high dose of unilateral and bilateral AAV2-82Q model showed a greater decrease in performance on the stepping and cylinder tests. We also observed more prominent EM48-positive mutant huntingtin aggregates in the medium spiny neurons of the high dose of AAV2-82Q injected group. Conclusion: Based on the results from the present study, high dose of AAV2-82Q is the optimum titer for establishing a Huntingtonrat model. Delivery of high dose of humanAAV2-82Q resulted in the manifestation of Huntington behaviors and optimum expression of the huntingtin protein in vivo.
Authors: Ana Fajardo-Serrano; Alberto J Rico; Elvira Roda; Adriana Honrubia; Sandra Arrieta; Goiaz Ariznabarreta; Julia Chocarro; Elena Lorenzo-Ramos; Alvaro Pejenaute; Alfonso Vázquez; José Luis Lanciego Journal: Biomedicines Date: 2022-03-23