CaltechAUTHORS
  A Caltech Library Service

Multicolor sparse viral labeling and 3D digital tracing of enteric plexus in mouse proximal colon using a novel adeno‐associated virus capsid

Wang, Lixin and Challis, Collin and Li, Songlin and Fowlkes, Charless C. and Ravindra Kumar, Sripriya and Yuan, Pu-Qing and Taché, Yvette F. (2020) Multicolor sparse viral labeling and 3D digital tracing of enteric plexus in mouse proximal colon using a novel adeno‐associated virus capsid. Neurogastroenterology and Motility . ISSN 1350-1925. (In Press) https://resolver.caltech.edu/CaltechAUTHORS:20201023-130934157

[img] PDF - Supplemental Material
See Usage Policy.

525Kb
[img] PDF (Fig. S1-S10) - Supplemental Material
See Usage Policy.

12Mb
[img] Video (MPEG) (Video S1) - Supplemental Material
See Usage Policy.

19Mb
[img] Video (MPEG) (Video S2) - Supplemental Material
See Usage Policy.

24Mb
[img] Video (MPEG) (Video S3) - Supplemental Material
See Usage Policy.

2314Kb

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20201023-130934157

Abstract

Background. Intravenous administration of adeno‐associated virus (AAV) can be used as a noninvasive approach to trace neuronal morphology and links. AAV‐PHP.S is a variant of AAV9 that effectively transduces the peripheral nervous system. The objective was to label randomly and sparsely enteric plexus in the mouse colon using AAV‐PHP.S with a tunable two‐component multicolor vector system and digitally trace individual neurons and nerve fibers within microcircuits in three dimensions (3D). Methods. A vector system including a tetracycline inducer with a tet‐responsive element driving three separate fluorophores was packaged in the AAV‐PHP.S capsid. The vectors were injected retro‐orbitally in mice, and the colon was harvested 3 weeks after. Confocal microscopic images of enteric plexus were digitally segmented and traced in 3D using Neurolucida 360, neuTube, or Imaris software. Key Results. The transduction of multicolor AAV vectors induced random sparse spectral labeling of soma and neurites primarily in the myenteric plexus of the proximal colon, while neurons in the submucosal plexus were occasionally transduced. Digital tracing in 3D showed various types of wiring, including multiple conjunctions of one neuron with other neurons, neurites en route, and endings; clusters of neurons in close apposition between each other; axon–axon parallel conjunctions; and intraganglionic nerve endings consisting of multiple nerve endings and passing fibers. Most of digitally traced neuronal somas were of small or medium in size. Conclusions & Inferences. The multicolor AAV‐PHP.S‐packaged vectors enabled random sparse spectral labeling and revealed complexities of enteric microcircuit in the mouse proximal colon. The techniques can facilitate digital modeling of enteric micro‐circuitry.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1111/nmo.14014DOIArticle
ORCID:
AuthorORCID
Wang, Lixin0000-0001-8140-6325
Challis, Collin0000-0003-4716-6086
Fowlkes, Charless C.0000-0002-2990-1780
Ravindra Kumar, Sripriya0000-0001-6033-7631
Yuan, Pu-Qing0000-0002-0067-510X
Taché, Yvette F.0000-0002-1044-5279
Additional Information:© 2020 John Wiley & Sons. Version of Record online: 23 October 2020. Manuscript accepted: 22 September 2020. Manuscript revised: 15 August 2020. Manuscript received: 24 September 2019. Funding information: NIH SPARC award OT2OD24899, Department of Veterans Affair Rehabilitation Research and Development grant 1RX001685, Veterans Administration Senior Research Career Scientist Award, P30 NIHDDK‐41301 UCLA/Digestive Diseases Research Core Center Grant.
Funders:
Funding AgencyGrant Number
NIHOT2OD24899
Department of Veterans Affairs1RX001685
NIHP30 NIHDDK‐41301
Record Number:CaltechAUTHORS:20201023-130934157
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201023-130934157
Official Citation:Wang L, Challis C, Li S, et al. Multicolor sparse viral labeling and 3D digital tracing of enteric plexus in mouse proximal colon using a novel adeno-associated virus capsid. Neurogastroenterology & Motility. 2020;00:e14014. https://doi.org/10.1111/nmo.14014
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:106263
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:23 Oct 2020 20:21
Last Modified:23 Oct 2020 20:21

Repository Staff Only: item control page