---
_id: '15016'
abstract:
- lang: eng
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. '
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)."
article_processing_charge: No
author:
- first_name: Eliza C.B.
full_name: Jaeger, Eliza C.B.
last_name: Jaeger
- first_name: David
full_name: Vijatovic, David
id: cf391e77-ec3c-11ea-a124-d69323410b58
last_name: Vijatovic
- first_name: Astrid
full_name: Deryckere, Astrid
last_name: Deryckere
- first_name: Nikol
full_name: Zorin, Nikol
last_name: Zorin
- first_name: Akemi L.
full_name: Nguyen, Akemi L.
last_name: Nguyen
- first_name: Georgiy
full_name: Ivanian, Georgiy
id: eaf2b366-cfd1-11ee-bbdf-c8790f800a05
last_name: Ivanian
- first_name: Jamie
full_name: Woych, Jamie
last_name: Woych
- first_name: Rebecca C
full_name: Arnold, Rebecca C
id: d6cce458-14c9-11ed-a755-c1c8fc6fde6f
last_name: Arnold
- first_name: Alonso
full_name: Ortega Gurrola, Alonso
last_name: Ortega Gurrola
- first_name: Arik
full_name: Shvartsman, Arik
last_name: Shvartsman
- first_name: Francesca
full_name: Barbieri, Francesca
id: a9492887-8972-11ed-ae7b-bfae10998254
last_name: Barbieri
- first_name: Florina-Alexandra
full_name: Toma, Florina-Alexandra
id: 85dd99f2-15b2-11ec-abd3-d1ae4d57f3b5
last_name: Toma
- first_name: Gary J.
full_name: Gorbsky, Gary J.
last_name: Gorbsky
- first_name: Marko E.
full_name: Horb, Marko E.
last_name: Horb
- first_name: Hollis T.
full_name: Cline, Hollis T.
last_name: Cline
- first_name: Timothy F.
full_name: Shay, Timothy F.
last_name: Shay
- first_name: Darcy B.
full_name: Kelley, Darcy B.
last_name: Kelley
- first_name: Ayako
full_name: Yamaguchi, Ayako
last_name: Yamaguchi
- first_name: Mark
full_name: Shein-Idelson, Mark
last_name: Shein-Idelson
- first_name: Maria Antonietta
full_name: Tosches, Maria Antonietta
last_name: Tosches
- first_name: Lora Beatrice Jaeger
full_name: Sweeney, Lora Beatrice Jaeger
id: 56BE8254-C4F0-11E9-8E45-0B23E6697425
last_name: Sweeney
orcid: 0000-0001-9242-5601
citation:
ama: Jaeger ECB, Vijatovic D, Deryckere A, et al. Adeno-associated viral tools to
trace neural development and connectivity across amphibians. bioRxiv. doi:10.1101/2024.02.15.580289
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. bioRxiv. https://doi.org/10.1101/2024.02.15.580289
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.” BioRxiv,
n.d. https://doi.org/10.1101/2024.02.15.580289.
ieee: E. C. B. Jaeger et al., “Adeno-associated viral tools to trace neural
development and connectivity across amphibians,” bioRxiv. .
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, 10.1101/2024.02.15.580289.
mla: Jaeger, Eliza C. B., et al. “Adeno-Associated Viral Tools to Trace Neural Development
and Connectivity across Amphibians.” BioRxiv, doi:10.1101/2024.02.15.580289.
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.).
date_created: 2024-02-20T09:20:32Z
date_published: 2024-02-16T00:00:00Z
date_updated: 2024-02-20T09:34:25Z
day: '16'
department:
- _id: LoSw
- _id: MaDe
- _id: GaNo
doi: 10.1101/2024.02.15.580289
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://doi.org/10.1101/2024.02.15.580289
month: '02'
oa: 1
oa_version: Preprint
project:
- _id: bd73af52-d553-11ed-ba76-912049f0ac7a
grant_number: FTI21-D-046
name: Entwicklung und Funktion der V1 Interneuronen vom Schwimmen zum Laufen während
der Metamorphose von Xenopus
- _id: ebb66355-77a9-11ec-83b8-b8ac210a4dae
grant_number: '101041551'
name: Development and Evolution of Tetrapod Motor Circuits
publication: bioRxiv
publication_status: submitted
status: public
title: Adeno-associated viral tools to trace neural development and connectivity across
amphibians
type: preprint
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
year: '2024'
...