A two-compartment organotypic model of mammalian peripheral nerve repair

Rezina Siddique, Alka Vyas, Nitish Thakor, Thomas M. Brushart

Research output: Contribution to journalArticle

Abstract

Background: Schwann cells in the distal stump of transected nerve upregulate growth factors that support regeneration on a modality-specific basis. It is unclear, however, which of these preferentially support motor axon regeneration. Identification of these factors will require a model that can isolate growth factor effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. New method: A two-compartment PDMS base is topped by a collagen-coated membrane that supports a spinal cord cross-section above one compartment. Fluorescent motoneurons in this section reinnervate a segment of peripheral nerve that directs axons through a water-tight barrier to the second compartment, where nerve repair is performed. Results: Motoneurons remain healthy for several weeks. The axons they project through the water-tight barrier survive transection and cross a nerve repair in substantial numbers to reinnervate an additional nerve segment. Fluidic isolation of the two compartments was confirmed with a dye leakage test, and the physiologic integrity of the system was tested by retrograde labeling of only those motor neurons to which tracer was exposed and by limitation of toxin effects to a single compartment. Comparison with existing methods: Nerve repair cannot be modeled in monolayer cell culture. Our previous organotypic model accurately modeled nerve repair, but did not allow individual control of motoneuron and growth cone environments. Conclusions: This model isolates treatment effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. Additionally, it facilitates surgical manipulation of tissues and high-resolution imaging.

Original languageEnglish (US)
Pages (from-to)84-92
Number of pages9
JournalJournal of Neuroscience Methods
Volume232
DOIs
StatePublished - Jul 30 2014

Fingerprint

Peripheral Nerves
Axons
Motor Neurons
Regeneration
Water
Growth Cones
Schwann Cells
Spinal Cord
Intercellular Signaling Peptides and Proteins
Coloring Agents
Up-Regulation
Collagen
Cell Culture Techniques
Membranes

Keywords

  • Organotypic
  • Peripheral nerve
  • Regeneration
  • Spinal cord
  • YFP

ASJC Scopus subject areas

  • Neuroscience(all)
  • Medicine(all)

Cite this

A two-compartment organotypic model of mammalian peripheral nerve repair. / Siddique, Rezina; Vyas, Alka; Thakor, Nitish; Brushart, Thomas M.

In: Journal of Neuroscience Methods, Vol. 232, 30.07.2014, p. 84-92.

Research output: Contribution to journalArticle

Siddique, Rezina; Vyas, Alka; Thakor, Nitish; Brushart, Thomas M. / A two-compartment organotypic model of mammalian peripheral nerve repair.

In: Journal of Neuroscience Methods, Vol. 232, 30.07.2014, p. 84-92.

Research output: Contribution to journalArticle

@article{12875abecb9047e7919666f0cd6bcc54,
title = "A two-compartment organotypic model of mammalian peripheral nerve repair",
abstract = "Background: Schwann cells in the distal stump of transected nerve upregulate growth factors that support regeneration on a modality-specific basis. It is unclear, however, which of these preferentially support motor axon regeneration. Identification of these factors will require a model that can isolate growth factor effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. New method: A two-compartment PDMS base is topped by a collagen-coated membrane that supports a spinal cord cross-section above one compartment. Fluorescent motoneurons in this section reinnervate a segment of peripheral nerve that directs axons through a water-tight barrier to the second compartment, where nerve repair is performed. Results: Motoneurons remain healthy for several weeks. The axons they project through the water-tight barrier survive transection and cross a nerve repair in substantial numbers to reinnervate an additional nerve segment. Fluidic isolation of the two compartments was confirmed with a dye leakage test, and the physiologic integrity of the system was tested by retrograde labeling of only those motor neurons to which tracer was exposed and by limitation of toxin effects to a single compartment. Comparison with existing methods: Nerve repair cannot be modeled in monolayer cell culture. Our previous organotypic model accurately modeled nerve repair, but did not allow individual control of motoneuron and growth cone environments. Conclusions: This model isolates treatment effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. Additionally, it facilitates surgical manipulation of tissues and high-resolution imaging.",
keywords = "Organotypic, Peripheral nerve, Regeneration, Spinal cord, YFP",
author = "Rezina Siddique and Alka Vyas and Nitish Thakor and Brushart, {Thomas M.}",
year = "2014",
month = "7",
doi = "10.1016/j.jneumeth.2014.05.005",
volume = "232",
pages = "84--92",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",

}

TY - JOUR

T1 - A two-compartment organotypic model of mammalian peripheral nerve repair

AU - Siddique,Rezina

AU - Vyas,Alka

AU - Thakor,Nitish

AU - Brushart,Thomas M.

PY - 2014/7/30

Y1 - 2014/7/30

N2 - Background: Schwann cells in the distal stump of transected nerve upregulate growth factors that support regeneration on a modality-specific basis. It is unclear, however, which of these preferentially support motor axon regeneration. Identification of these factors will require a model that can isolate growth factor effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. New method: A two-compartment PDMS base is topped by a collagen-coated membrane that supports a spinal cord cross-section above one compartment. Fluorescent motoneurons in this section reinnervate a segment of peripheral nerve that directs axons through a water-tight barrier to the second compartment, where nerve repair is performed. Results: Motoneurons remain healthy for several weeks. The axons they project through the water-tight barrier survive transection and cross a nerve repair in substantial numbers to reinnervate an additional nerve segment. Fluidic isolation of the two compartments was confirmed with a dye leakage test, and the physiologic integrity of the system was tested by retrograde labeling of only those motor neurons to which tracer was exposed and by limitation of toxin effects to a single compartment. Comparison with existing methods: Nerve repair cannot be modeled in monolayer cell culture. Our previous organotypic model accurately modeled nerve repair, but did not allow individual control of motoneuron and growth cone environments. Conclusions: This model isolates treatment effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. Additionally, it facilitates surgical manipulation of tissues and high-resolution imaging.

AB - Background: Schwann cells in the distal stump of transected nerve upregulate growth factors that support regeneration on a modality-specific basis. It is unclear, however, which of these preferentially support motor axon regeneration. Identification of these factors will require a model that can isolate growth factor effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. New method: A two-compartment PDMS base is topped by a collagen-coated membrane that supports a spinal cord cross-section above one compartment. Fluorescent motoneurons in this section reinnervate a segment of peripheral nerve that directs axons through a water-tight barrier to the second compartment, where nerve repair is performed. Results: Motoneurons remain healthy for several weeks. The axons they project through the water-tight barrier survive transection and cross a nerve repair in substantial numbers to reinnervate an additional nerve segment. Fluidic isolation of the two compartments was confirmed with a dye leakage test, and the physiologic integrity of the system was tested by retrograde labeling of only those motor neurons to which tracer was exposed and by limitation of toxin effects to a single compartment. Comparison with existing methods: Nerve repair cannot be modeled in monolayer cell culture. Our previous organotypic model accurately modeled nerve repair, but did not allow individual control of motoneuron and growth cone environments. Conclusions: This model isolates treatment effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. Additionally, it facilitates surgical manipulation of tissues and high-resolution imaging.

KW - Organotypic

KW - Peripheral nerve

KW - Regeneration

KW - Spinal cord

KW - YFP

UR - http://www.scopus.com/inward/record.url?scp=84901477199&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84901477199&partnerID=8YFLogxK

U2 - 10.1016/j.jneumeth.2014.05.005

DO - 10.1016/j.jneumeth.2014.05.005

M3 - Article

VL - 232

SP - 84

EP - 92

JO - Journal of Neuroscience Methods

T2 - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

ER -