Retinal cells produce TIMP-1 and TIMP-2 in response to cyclic mechanical stretching

Masako Namba, Toshihiko Matsuo, Fumio Shiraga, Hiroshi Ohtsuki

Research output: Contribution to journalArticle

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Abstract

Purpose: Tissue inhibitors of metalloproteinases (TIMPs) are crucial for the maintenance of retinal extracellular matrix such as interphotoreceptor matrix and internal limiting membrane. This study is to examine whether retinal cells respond to mechanical stretching and produce TIMPs. Methods: Chick retinal adherent cells in near confluency were exposed to mechanical stretching of the bottom of a 6-cm Petri dish at the maximum magnitude of 4,500 microstrain and at a cycle of 30 s for 72 h. TIMP-1 and TIMP-2 levels in the medium at 24, 48 and 72 h after the beginning of stretching were measured by enzyme immunoassay, and their expression was examined by immunohistochemistry. Results: The number of retinal cells during the 72-hour period of stretching did not change significantly both in the stretched group and in the nonstretched control group. Retinal cells in the stretched group produced significantly larger amounts of TIMP-1 and TIMP-2 at 48 h after stretching, compared with nonstretched controls (p = 0.0163 and p = 0.047, respectively, Mann-Whitney U test). Immunohistochemically, a large part of retinal cells in nonstretched Petri dishes was positive for glial fibrillary acidic protein, indicative of glial cells, while some small foci of cells were positive for neuron-specific enolase, indicative of neurons. Fluorescent double labeling demonstrated that both glial cells and neurons were positive for TIMP-1 and TIMP-2. Conclusion: Chick retinal cells, most of which were glial cells mixed with a small number of neurons, produced TIMP-1 and TIMP-2. Their production was enhanced by cyclic mechanical stretching.

Original languageEnglish
Pages (from-to)163-169
Number of pages7
JournalOphthalmic Research
Volume33
Issue number3
DOIs
Publication statusPublished - 2001

Fingerprint

Tissue Inhibitor of Metalloproteinase-2
Tissue Inhibitor of Metalloproteinase-1
Neuroglia
Tissue Inhibitor of Metalloproteinases
Neurons
Phosphopyruvate Hydratase
Glial Fibrillary Acidic Protein
Nonparametric Statistics
Immunoenzyme Techniques
Extracellular Matrix
Cell Count
Immunohistochemistry
Maintenance
Control Groups
Membranes

Keywords

  • Cyclic mechanical stretching
  • Retinal (glial) cells
  • Tissue inhibitors of metalloproteinases (TIMP)

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Retinal cells produce TIMP-1 and TIMP-2 in response to cyclic mechanical stretching. / Namba, Masako; Matsuo, Toshihiko; Shiraga, Fumio; Ohtsuki, Hiroshi.

In: Ophthalmic Research, Vol. 33, No. 3, 2001, p. 163-169.

Research output: Contribution to journalArticle

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AU - Shiraga, Fumio

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AB - Purpose: Tissue inhibitors of metalloproteinases (TIMPs) are crucial for the maintenance of retinal extracellular matrix such as interphotoreceptor matrix and internal limiting membrane. This study is to examine whether retinal cells respond to mechanical stretching and produce TIMPs. Methods: Chick retinal adherent cells in near confluency were exposed to mechanical stretching of the bottom of a 6-cm Petri dish at the maximum magnitude of 4,500 microstrain and at a cycle of 30 s for 72 h. TIMP-1 and TIMP-2 levels in the medium at 24, 48 and 72 h after the beginning of stretching were measured by enzyme immunoassay, and their expression was examined by immunohistochemistry. Results: The number of retinal cells during the 72-hour period of stretching did not change significantly both in the stretched group and in the nonstretched control group. Retinal cells in the stretched group produced significantly larger amounts of TIMP-1 and TIMP-2 at 48 h after stretching, compared with nonstretched controls (p = 0.0163 and p = 0.047, respectively, Mann-Whitney U test). Immunohistochemically, a large part of retinal cells in nonstretched Petri dishes was positive for glial fibrillary acidic protein, indicative of glial cells, while some small foci of cells were positive for neuron-specific enolase, indicative of neurons. Fluorescent double labeling demonstrated that both glial cells and neurons were positive for TIMP-1 and TIMP-2. Conclusion: Chick retinal cells, most of which were glial cells mixed with a small number of neurons, produced TIMP-1 and TIMP-2. Their production was enhanced by cyclic mechanical stretching.

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