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A method to stretch the retina in vitro and monitor effects on Müller glial cells

Lindqvist, Niclas LU ; Liu, Q; Zajadacz, Joachim; Franze, Kristian and Reichenbach, Andreas (2008) 38-th Annual Meeting, Society for Neuroscience
Abstract (Swedish)
The retina is a thin tissue layer on the inside of the eye and may be subject to tractional forces in diseases like myopia, retinoschisis, and retinal detachment. Müller glial cells (MGCs) are the only cells spanning the entire retinal thickness and hypothetically respond to mechanical stress imposed across retinal layers. We designed a device to apply force in vitro perpendicular to the retinal surface thereby stretching the retinal layers and the MGCs. A slice of retina, mounted between filters, was positioned between two hollow metallic tubes through which vacuum pressure was applied. The tubes met inside a buffer-filled container, one of them was fixed and the other was movable using a micrometer-stepping motor. The device was fitted... (More)
The retina is a thin tissue layer on the inside of the eye and may be subject to tractional forces in diseases like myopia, retinoschisis, and retinal detachment. Müller glial cells (MGCs) are the only cells spanning the entire retinal thickness and hypothetically respond to mechanical stress imposed across retinal layers. We designed a device to apply force in vitro perpendicular to the retinal surface thereby stretching the retinal layers and the MGCs. A slice of retina, mounted between filters, was positioned between two hollow metallic tubes through which vacuum pressure was applied. The tubes met inside a buffer-filled container, one of them was fixed and the other was movable using a micrometer-stepping motor. The device was fitted onto a microscope stage to monitor the retina in cross-section during stretch. First, we used guinea pig retinae with clearly distinguishable MGC processes. MGCs were selectively stained using the calcium-sensitive, fluorescent dye X-rhod-1. During stretch, we recorded an acute increase in fluorescence in MGCs indicative of increased intracellular calcium levels.
Further, we used rat retinae to study if a short period of stretching led to a subsequent activation of intracellular pathways. Using immunohistochemical detection we found a fast activation of extracellular regulated kinase (ERK) preferentially in the MGCs. One hour after stretch, the transcription factor c-fos was significantly upregulated mainly in the MGCs indicating stretch effects on gene transcription. Finally, we analyzed basic fibroblast growth factor expression, and found significantly increased expression in MGCs a few hours after stretch. In conclusion, we have developed a technique for real-time monitoring of MGCs during retinal stretch and we have identified the MGCs to be mechano-responsive cells in the retina. (Less)
Abstract
The retina is a thin tissue layer on the inside of the eye and may be subject to tractional forces in diseases like myopia, retinoschisis, and retinal detachment. Müller glial cells (MGCs) are the only cells spanning the entire retinal thickness and hypothetically respond to mechanical stress imposed across retinal layers. We designed a device to apply force in vitro perpendicular to the retinal surface thereby stretching the retinal layers and the MGCs. A slice of retina, mounted between filters, was positioned between two hollow metallic tubes through which vacuum pressure was applied. The tubes met inside a buffer-filled container, one of them was fixed and the other was movable using a micrometer-stepping motor. The device was fitted... (More)
The retina is a thin tissue layer on the inside of the eye and may be subject to tractional forces in diseases like myopia, retinoschisis, and retinal detachment. Müller glial cells (MGCs) are the only cells spanning the entire retinal thickness and hypothetically respond to mechanical stress imposed across retinal layers. We designed a device to apply force in vitro perpendicular to the retinal surface thereby stretching the retinal layers and the MGCs. A slice of retina, mounted between filters, was positioned between two hollow metallic tubes through which vacuum pressure was applied. The tubes met inside a buffer-filled container, one of them was fixed and the other was movable using a micrometer-stepping motor. The device was fitted onto a microscope stage to monitor the retina in cross-section during stretch. First, we used guinea pig retinae with clearly distinguishable MGC processes. MGCs were selectively stained using the calcium-sensitive, fluorescent dye X-rhod-1. During stretch, we recorded an acute increase in fluorescence in MGCs indicative of increased intracellular calcium levels.
Further, we used rat retinae to study if a short period of stretching led to a subsequent activation of intracellular pathways. Using immunohistochemical detection we found a fast activation of extracellular regulated kinase (ERK) preferentially in the MGCs. One hour after stretch, the transcription factor c-fos was significantly upregulated mainly in the MGCs indicating stretch effects on gene transcription. Finally, we analyzed basic fibroblast growth factor expression, and found significantly increased expression in MGCs a few hours after stretch. In conclusion, we have developed a technique for real-time monitoring of MGCs during retinal stretch and we have identified the MGCs to be mechano-responsive cells in the retina. (Less)
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38-th Annual Meeting, Society for Neuroscience
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@misc{684c0768-aad7-4047-a81d-4a29fa812df2,
  abstract     = {The retina is a thin tissue layer on the inside of the eye and may be subject to tractional forces in diseases like myopia, retinoschisis, and retinal detachment. Müller glial cells (MGCs) are the only cells spanning the entire retinal thickness and hypothetically respond to mechanical stress imposed across retinal layers. We designed a device to apply force in vitro perpendicular to the retinal surface thereby stretching the retinal layers and the MGCs. A slice of retina, mounted between filters, was positioned between two hollow metallic tubes through which vacuum pressure was applied. The tubes met inside a buffer-filled container, one of them was fixed and the other was movable using a micrometer-stepping motor. The device was fitted onto a microscope stage to monitor the retina in cross-section during stretch. First, we used guinea pig retinae with clearly distinguishable MGC processes. MGCs were selectively stained using the calcium-sensitive, fluorescent dye X-rhod-1. During stretch, we recorded an acute increase in fluorescence in MGCs indicative of increased intracellular calcium levels.<br/>Further, we used rat retinae to study if a short period of stretching led to a subsequent activation of intracellular pathways. Using immunohistochemical detection we found a fast activation of extracellular regulated kinase (ERK) preferentially in the MGCs. One hour after stretch, the transcription factor c-fos was significantly upregulated mainly in the MGCs indicating stretch effects on gene transcription. Finally, we analyzed basic fibroblast growth factor expression, and found significantly increased expression in MGCs a few hours after stretch. In conclusion, we have developed a technique for real-time monitoring of MGCs during retinal stretch and we have identified the MGCs to be mechano-responsive cells in the retina.},
  author       = {Lindqvist, Niclas and Liu, Q and Zajadacz, Joachim and Franze, Kristian and Reichenbach, Andreas},
  language     = {eng},
  title        = {A method to stretch the retina in vitro and monitor effects on Müller glial cells},
  year         = {2008},
}