{"main_file_link":[{"open_access":"1","url":"https://doi.org/10.1101/2024.02.15.580289"}],"author":[{"full_name":"Jaeger, Eliza C.B.","first_name":"Eliza C.B.","last_name":"Jaeger"},{"full_name":"Vijatovic, David","id":"cf391e77-ec3c-11ea-a124-d69323410b58","first_name":"David","last_name":"Vijatovic"},{"full_name":"Deryckere, Astrid","last_name":"Deryckere","first_name":"Astrid"},{"last_name":"Zorin","first_name":"Nikol","full_name":"Zorin, Nikol"},{"last_name":"Nguyen","first_name":"Akemi L.","full_name":"Nguyen, Akemi L."},{"id":"eaf2b366-cfd1-11ee-bbdf-c8790f800a05","full_name":"Ivanian, Georgiy","last_name":"Ivanian","first_name":"Georgiy"},{"full_name":"Woych, Jamie","last_name":"Woych","first_name":"Jamie"},{"last_name":"Arnold","first_name":"Rebecca C","id":"d6cce458-14c9-11ed-a755-c1c8fc6fde6f","full_name":"Arnold, Rebecca C"},{"full_name":"Ortega Gurrola, Alonso","first_name":"Alonso","last_name":"Ortega Gurrola"},{"first_name":"Arik","last_name":"Shvartsman","full_name":"Shvartsman, Arik"},{"id":"a9492887-8972-11ed-ae7b-bfae10998254","full_name":"Barbieri, Francesca","last_name":"Barbieri","first_name":"Francesca"},{"id":"85dd99f2-15b2-11ec-abd3-d1ae4d57f3b5","full_name":"Toma, Florina-Alexandra","last_name":"Toma","first_name":"Florina-Alexandra"},{"full_name":"Gorbsky, Gary J.","first_name":"Gary J.","last_name":"Gorbsky"},{"last_name":"Horb","first_name":"Marko E.","full_name":"Horb, Marko E."},{"full_name":"Cline, Hollis T.","last_name":"Cline","first_name":"Hollis T."},{"full_name":"Shay, Timothy F.","last_name":"Shay","first_name":"Timothy F."},{"full_name":"Kelley, Darcy B.","last_name":"Kelley","first_name":"Darcy B."},{"last_name":"Yamaguchi","first_name":"Ayako","full_name":"Yamaguchi, Ayako"},{"first_name":"Mark","last_name":"Shein-Idelson","full_name":"Shein-Idelson, Mark"},{"full_name":"Tosches, Maria Antonietta","last_name":"Tosches","first_name":"Maria Antonietta"},{"first_name":"Lora Beatrice Jaeger","last_name":"Sweeney","orcid":"0000-0001-9242-5601","full_name":"Sweeney, Lora Beatrice Jaeger","id":"56BE8254-C4F0-11E9-8E45-0B23E6697425"}],"publication":"bioRxiv","day":"16","language":[{"iso":"eng"}],"year":"2024","oa":1,"abstract":[{"text":"The development, evolution, and function of the vertebrate central nervous system (CNS) can be best studied using diverse model organisms. Amphibians, with their unique phylogenetic position at the transition between aquatic and terrestrial lifestyles, are valuable for understanding the origin and evolution of the tetrapod brain and spinal cord. Their metamorphic developmental transitions and unique regenerative abilities also facilitate the discovery of mechanisms for neural circuit remodeling and replacement. The genetic toolkit for amphibians, however, remains limited, with only a few species having sequenced genomes and a small number of transgenic lines available. In mammals, recombinant adeno-associated viral vectors (AAVs) have become a powerful alternative to genome modification for visualizing and perturbing the nervous system. AAVs are DNA viruses that enable neuronal transduction in both developing and adult animals with low toxicity and spatial, temporal, and cell-type specificity. However, AAVs have never been shown to transduce amphibian cells efficiently. To bridge this gap, we established a simple, scalable, and robust strategy to screen AAV serotypes in three distantly-related amphibian species: the frogs Xenopus laevis and Pelophylax bedriagae, and the salamander Pleurodeles waltl, in both developing larval tadpoles and post-metamorphic animals. For each species, we successfully identified at least two AAV serotypes capable of infecting the CNS; however, no pan-amphibian serotype was identified, indicating rapid evolution of AAV tropism. In addition, we developed an AAV-based strategy that targets isochronic cohorts of developing neurons – a critical tool for parsing neural circuit assembly. Finally, to enable visualization and manipulation of neural circuits, we identified AAV variants for retrograde tracing of neuronal projections in adult animals. Our findings expand the toolkit for amphibians to include AAVs, establish a generalizable workflow for AAV screening in non-canonical research organisms, generate testable hypotheses for the evolution of AAV tropism, and lay the foundation for modern cross-species comparisons of vertebrate CNS development, function, and evolution. ","lang":"eng"}],"publication_status":"submitted","date_created":"2024-02-20T09:20:32Z","month":"02","_id":"15016","status":"public","date_published":"2024-02-16T00:00:00Z","type":"preprint","date_updated":"2024-02-20T09:34:25Z","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","project":[{"_id":"bd73af52-d553-11ed-ba76-912049f0ac7a","name":"Entwicklung und Funktion der V1 Interneuronen vom Schwimmen zum Laufen während der Metamorphose von Xenopus","grant_number":"FTI21-D-046"},{"_id":"ebb66355-77a9-11ec-83b8-b8ac210a4dae","name":"Development and Evolution of Tetrapod Motor Circuits","grant_number":"101041551"}],"oa_version":"Preprint","department":[{"_id":"LoSw"},{"_id":"MaDe"},{"_id":"GaNo"}],"doi":"10.1101/2024.02.15.580289","acknowledgement":"We would like to extend our thanks to members of the Sweeney, Tosches, Shein-Idelson,\r\nYamaguchi, Kelley, and Cline Labs for their contributions to this project, discussion and support.\r\nWe additionally thank the Beckman Institute Clover Center and Viviana Gradinaru (Caltech),\r\nKimberly Ritola (UNC NeuroTools), Flavia Gama Gomez Leite (ISTA Viral Core), and Hüseyin\r\nCihan Önal (Shigemoto Group, ISTA) for their consultation and assistance regarding AAVs, as\r\nwell as Andras Simon and Alberto Joven for feedback and discussions on AAVs in Pleurodeles.\r\nTo do these experiments, we have also benefited from the tremendous support of our animal care and imaging facilities at our respective institutions, as well as the amphibian stock centers\r\n(National Xenopus Resource Center, European Xenopus Resource Center, Xenopus Express)\r\nand our funding sources: U.S. National Science Foundation (NSF) Grant Number IOS 2110086\r\n(D.B.K., L.B.S., M.A.T., A.Y., and H.T.C.); United States-Israel Binational Science Foundation\r\n(BSF) Grant Number 2020702 (M.S.-I.); NSF Award Number 1645105 (G.J.G., M.E.H.); FTI\r\nStrategy Lower Austria Dissertation Grant Number FTI21-D-046 (D.V.); Horizon Europe ERC\r\nStarting Grant Number 101041551 (L.B.S.); NIH grant number R35GM146973 (M.A.T.); Rita Allen\r\nFoundation award number GA_032522_FE (M.A.T.); European Molecular Biology Organization\r\nLong-Term Fellowship ALTF 874-2021 (A.D.); National Science Foundation Graduate Research\r\nFellowship DGE 2036197 (E.C.J.B.); NIH grant number P40OD010997 (M.E.H).","title":"Adeno-associated viral tools to trace neural development and connectivity across amphibians","citation":{"short":"E.C.B. Jaeger, D. Vijatovic, A. Deryckere, N. Zorin, A.L. Nguyen, G. Ivanian, J. Woych, R.C. Arnold, A. Ortega Gurrola, A. Shvartsman, F. Barbieri, F.-A. Toma, G.J. Gorbsky, M.E. Horb, H.T. Cline, T.F. Shay, D.B. Kelley, A. Yamaguchi, M. Shein-Idelson, M.A. Tosches, L.B. Sweeney, BioRxiv (n.d.).","ama":"Jaeger ECB, Vijatovic D, Deryckere A, et al. Adeno-associated viral tools to trace neural development and connectivity across amphibians. <i>bioRxiv</i>. doi:<a href=\"https://doi.org/10.1101/2024.02.15.580289\">10.1101/2024.02.15.580289</a>","chicago":"Jaeger, Eliza C.B., David Vijatovic, Astrid Deryckere, Nikol Zorin, Akemi L. Nguyen, Georgiy Ivanian, Jamie Woych, et al. “Adeno-Associated Viral Tools to Trace Neural Development and Connectivity across Amphibians.” <i>BioRxiv</i>, n.d. <a href=\"https://doi.org/10.1101/2024.02.15.580289\">https://doi.org/10.1101/2024.02.15.580289</a>.","apa":"Jaeger, E. C. B., Vijatovic, D., Deryckere, A., Zorin, N., Nguyen, A. L., Ivanian, G., … Sweeney, L. B. (n.d.). Adeno-associated viral tools to trace neural development and connectivity across amphibians. <i>bioRxiv</i>. <a href=\"https://doi.org/10.1101/2024.02.15.580289\">https://doi.org/10.1101/2024.02.15.580289</a>","ieee":"E. C. B. Jaeger <i>et al.</i>, “Adeno-associated viral tools to trace neural development and connectivity across amphibians,” <i>bioRxiv</i>. .","ista":"Jaeger ECB, Vijatovic D, Deryckere A, Zorin N, Nguyen AL, Ivanian G, Woych J, Arnold RC, Ortega Gurrola A, Shvartsman A, Barbieri F, Toma F-A, Gorbsky GJ, Horb ME, Cline HT, Shay TF, Kelley DB, Yamaguchi A, Shein-Idelson M, Tosches MA, Sweeney LB. Adeno-associated viral tools to trace neural development and connectivity across amphibians. bioRxiv, <a href=\"https://doi.org/10.1101/2024.02.15.580289\">10.1101/2024.02.15.580289</a>.","mla":"Jaeger, Eliza C. B., et al. “Adeno-Associated Viral Tools to Trace Neural Development and Connectivity across Amphibians.” <i>BioRxiv</i>, doi:<a href=\"https://doi.org/10.1101/2024.02.15.580289\">10.1101/2024.02.15.580289</a>."},"article_processing_charge":"No"}