Advanced

Neuronal integration in an abutting-retinas culture system

Zhang, Yiqin LU ; Caffé, A Romeo; Azadi, Seifollah LU ; van Veen, Theo LU ; Ehinger, Berndt LU and Perez, Maria-Thereza R LU (2003) In Investigative Ophthalmology & Visual Science 44(11). p.4936-4946
Abstract

PURPOSE: Limited integration is consistently observed between subretinal transplants and host retinas. In the current study, an in vitro model system for studying connections forming between two abutting retinas was developed.

METHODS: Neuroretinas were dissected from normal wild-type (WT) mice and green fluorescent protein (GFP) transgenic mice (obtained at postnatal days [P]0, P5, or P60), as well as from adult rd mice. Pieces from two different retinas (WT-WT, GFP-WT, GFP-rd) were placed side-by-side (contacting each other at the margins) or overlapping each other in organ cultures for 7 or 12 days. The abutting retinal pieces derived from animals of the same age (P5-P5; P60-P60) or of different ages (P0-P60; P5-P60). Retinal... (More)

PURPOSE: Limited integration is consistently observed between subretinal transplants and host retinas. In the current study, an in vitro model system for studying connections forming between two abutting retinas was developed.

METHODS: Neuroretinas were dissected from normal wild-type (WT) mice and green fluorescent protein (GFP) transgenic mice (obtained at postnatal days [P]0, P5, or P60), as well as from adult rd mice. Pieces from two different retinas (WT-WT, GFP-WT, GFP-rd) were placed side-by-side (contacting each other at the margins) or overlapping each other in organ cultures for 7 or 12 days. The abutting retinal pieces derived from animals of the same age (P5-P5; P60-P60) or of different ages (P0-P60; P5-P60). Retinal cells and fibers were visualized in wholemount preparations and in cross sections by immunocytochemistry using antibodies against neurofilament (NF+), neuronal nitric oxide synthase (NOS+), and protein kinase C (PKC+) and by GFP fluorescence (GFP+).

RESULTS: In side-by-side pairs (WT-WT, GFP-WT), numerous horizontal cell fibers (NF+) and amacrine cell fibers (NOS+) crossed the interface between the two pieces, forming continuous plexiform layers. In overlapping pairs, NF+, NOS+, and PKC+ fibers displayed parallel plexiform layers, and no crossover of fibers was observed in any of the pair combinations examined (WT-WT, GFP-WT, GFP-rd). Some integration was seen only in small areas where the structure of both retinal pieces was disrupted at the interface.

CONCLUSIONS: The results demonstrate the ability of neurites to extend between abutting retinas and to make appropriate target choices when they are placed side-by-side. However, this ability is limited when they overlap each other, similar to that observed in subretinal transplantation.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, Fluorescent Antibody Technique, Indirect, Green Fluorescent Proteins, Indicators and Reagents, Luminescent Proteins, Mice, Mice, Inbred C3H, Mice, Inbred C57BL, Mice, Transgenic, Neural Pathways, Neurites, Neurofilament Proteins, Neuroglia, Nitric Oxide Synthase, Nitric Oxide Synthase Type I, Organ Culture Techniques, Protein Kinase C, Retina, Retinal Degeneration, Journal Article, Research Support, Non-U.S. Gov't
in
Investigative Ophthalmology & Visual Science
volume
44
issue
11
pages
11 pages
publisher
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
external identifiers
  • pmid:14578420
  • wos:000186231800047
  • scopus:0142169869
ISSN
1552-5783
DOI
10.1167/iovs.02-0640
language
English
LU publication?
yes
id
40a44c28-ef84-44ce-8897-b6da81e4dd1b (old id 296467)
date added to LUP
2007-09-24 15:47:39
date last changed
2018-01-07 09:05:14
@article{40a44c28-ef84-44ce-8897-b6da81e4dd1b,
  abstract     = {<p>PURPOSE: Limited integration is consistently observed between subretinal transplants and host retinas. In the current study, an in vitro model system for studying connections forming between two abutting retinas was developed.</p><p>METHODS: Neuroretinas were dissected from normal wild-type (WT) mice and green fluorescent protein (GFP) transgenic mice (obtained at postnatal days [P]0, P5, or P60), as well as from adult rd mice. Pieces from two different retinas (WT-WT, GFP-WT, GFP-rd) were placed side-by-side (contacting each other at the margins) or overlapping each other in organ cultures for 7 or 12 days. The abutting retinal pieces derived from animals of the same age (P5-P5; P60-P60) or of different ages (P0-P60; P5-P60). Retinal cells and fibers were visualized in wholemount preparations and in cross sections by immunocytochemistry using antibodies against neurofilament (NF+), neuronal nitric oxide synthase (NOS+), and protein kinase C (PKC+) and by GFP fluorescence (GFP+).</p><p>RESULTS: In side-by-side pairs (WT-WT, GFP-WT), numerous horizontal cell fibers (NF+) and amacrine cell fibers (NOS+) crossed the interface between the two pieces, forming continuous plexiform layers. In overlapping pairs, NF+, NOS+, and PKC+ fibers displayed parallel plexiform layers, and no crossover of fibers was observed in any of the pair combinations examined (WT-WT, GFP-WT, GFP-rd). Some integration was seen only in small areas where the structure of both retinal pieces was disrupted at the interface.</p><p>CONCLUSIONS: The results demonstrate the ability of neurites to extend between abutting retinas and to make appropriate target choices when they are placed side-by-side. However, this ability is limited when they overlap each other, similar to that observed in subretinal transplantation.</p>},
  author       = {Zhang, Yiqin and Caffé, A Romeo and Azadi, Seifollah and van Veen, Theo and Ehinger, Berndt and Perez, Maria-Thereza R},
  issn         = {1552-5783},
  keyword      = {Animals,Fluorescent Antibody Technique, Indirect,Green Fluorescent Proteins,Indicators and Reagents,Luminescent Proteins,Mice,Mice, Inbred C3H,Mice, Inbred C57BL,Mice, Transgenic,Neural Pathways,Neurites,Neurofilament Proteins,Neuroglia,Nitric Oxide Synthase,Nitric Oxide Synthase Type I,Organ Culture Techniques,Protein Kinase C,Retina,Retinal Degeneration,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  number       = {11},
  pages        = {4936--4946},
  publisher    = {ASSOC RESEARCH VISION OPHTHALMOLOGY INC},
  series       = {Investigative Ophthalmology & Visual Science},
  title        = {Neuronal integration in an abutting-retinas culture system},
  url          = {http://dx.doi.org/10.1167/iovs.02-0640},
  volume       = {44},
  year         = {2003},
}