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Mitsuru Arima, Shigeo Yoshida, Keijiro Ishikawa, Shintaro Nakao, Ryo Asato, Yukio Sassa, Takeshi Kita, Akira Kudo, Akira Matsuda, Tatsuro Ishibashi; The Role Of Periostin In The Regression Of Hyaloid Vascular System During Ocular Development. Invest. Ophthalmol. Vis. Sci. 2012;53(14):5123.
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The appropriate regression of hyaloid vascular system (HVS) is required for normal ocular development and the failure of the regression could cause severe visual loss. Although it is recognized vitreous macrophage lineage cells play a crucial role in this process, the accurate mechanism is unclear. Periostin is a matricellular protein that is involved in tissue and vascular remodeling. The purpose of this study was to investigate the role of periostin in the HVS regression.
Wild type and periostin null mice eyeballs were enucleated at postnatal day (P) 4, 5, 7, 9, 12 and 15. Hyaloid vessels and vitreous macrophages in flat-mounted ocular specimen at each time point were stained with lectin B4 or with F4/80 or Iba-1, respectively. For quantification of the apoptosis of endothelial cells of hyaloid vessels, TUNEL labeling was performed. Immunohistochemistry was performed to detect the localization of periostin. Laser capture microdissection (LCM) and semiquantitative RT-PCR was also performed to determine the levels of periostin mRNA.
Lectin flat mount staining revealed the pronounced delay of HVS regression in periostin null mice relative to wild type. However the total number of F4/80 positive cells significantly increased in periostin null mice at each time point examined. The number of TUNEL positive endothelial cells reached a peak at P7 in wild type and P12 in periostin null mice, respectively. Additionally, the only small population of Iba-1 positive cells in vitreous cavity, which located nearby hyaloid vessels, was costained with periostin. Immunofluorescent triple staining of acridine orange, F4/80 and periostin revealed that acridine orange staining colocalized only with F4/80 but not with periostin. RT-PCR of vitreous macrophage lineage cells collected by LCM revealed the production of periostin mRNA.
These results suggest that periostin is produced by the macrophages activated in the vitreous cavity and may be involved in the HVS regression.
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