دورية أكاديمية
أعرض في EDS

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

التفاصيل البيبلوغرافية
العنوان: Multicolor sparse viral labeling and 3D digital tracing of enteric plexus in mouse proximal colon using a novel adeno-associated virus capsid.
المؤلفون: Wang L; Department of Medicine, Taman Manoukisan Digestive Diseases Division, UCLA, Los Angeles, CA, USA.; Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, CA, USA., Challis C; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Li S; Department of Medicine, Taman Manoukisan Digestive Diseases Division, UCLA, Los Angeles, CA, USA., Fowlkes CC; Department of Computer Science, University of California, Irvine, CA, USA., Ravindra Kumar S; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Yuan PQ; Department of Medicine, Taman Manoukisan Digestive Diseases Division, UCLA, Los Angeles, CA, USA., Taché YF; Department of Medicine, Taman Manoukisan Digestive Diseases Division, UCLA, Los Angeles, CA, USA.; Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, CA, USA.
المصدر: Neurogastroenterology and motility [Neurogastroenterol Motil] 2021 Aug; Vol. 33 (8), pp. e14014. Date of Electronic Publication: 2020 Oct 23.
نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.
اللغة: English
بيانات الدورية: Publisher: Blackwell Scientific Publications Country of Publication: England NLM ID: 9432572 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1365-2982 (Electronic) Linking ISSN: 13501925 NLM ISO Abbreviation: Neurogastroenterol Motil Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Osney Mead, Oxford, UK : Blackwell Scientific Publications, c1994-
مواضيع طبية MeSH: Colon/*metabolism , Enteric Nervous System/*metabolism , Submucous Plexus/*metabolism, Animals ; Colon/innervation ; Dependovirus ; Enteric Nervous System/virology ; Female ; Gene Transfer Techniques ; Green Fluorescent Proteins ; Male ; Mice ; Submucous Plexus/virology
مستخلص: 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.
(© 2020 John Wiley & Sons Ltd.)
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معلومات مُعتمدة: P30 DK041301 United States DK NIDDK NIH HHS; OT2 OD024899 United States OD NIH HHS; I01 RX001685 United States RX RRD VA
فهرسة مساهمة: Keywords: 3D imaging; adeno-associated virus; colonic enteric neuron; digital tracing; mouse
المشرفين على المادة: 147336-22-9 (Green Fluorescent Proteins)
تواريخ الأحداث: Date Created: 20201023 Date Completed: 20220124 Latest Revision: 20220124
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC8568587
DOI: 10.1111/nmo.14014
PMID: 33094876
قاعدة البيانات: MEDLINE
الوصف
تدمد:1365-2982
DOI:10.1111/nmo.14014