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Kenneth M. Yee, Fred N. Ross-Cisneros, Bridget N. Ballard, Michele C. Madigan, Jan M. Provis, Lloyd P. Aiello, Alfredo A. Sadun, Jerry Sebag; Immunohistochemistry of Human Embryonic Vitreous. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4106.
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© ARVO (1962-2015); The Authors (2016-present)
Previous studies (Feener, ARVO 2010) characterized the proteomic profile of embryonic human vitreous and detected a significant (2 to 5-fold) change in protein concentrations during the 2nd trimester. The present study examined 5 of these proteins for which antibodies were commercially available to confirm their presence and to characterize the location of expression in or adjacent to vitreous. Differences from early to late 2nd trimester were also determined.
A formalin fixed eye was obtained from 10, 14, and 18 gestational week old (GW; determined by ultrasound) human embryos. Three membrane (Cadherin, Dystroglycan and Clusterin), one secretory (Cystatin), and one cytoplasmic protein (Profilin-1) were studied by immunoperoxidase staining with antigen retrieval and hematoxylin counterstain.
Four proteins stained positively in vitreous. No staining was observed for Cystatin. At all ages, Dystroglycan was found on the cell membranes of hyalocytes and endothelial cells of the hyaloid artery (HA) and tunica vasculosa lentis (TVL). Profilin-1 was seen in the cytoplasm of these cells. There were age-related differences in Clusterin, which stained positively in hyalocytes of the 10 and 14GW eyes, but not the HA or TVL. In the 18GW eye, Clusterin was apparent in the HA. Similarly, Cadherin did not stain the HA of the 10GW eye, but was observed in vessels at 14 and 18GW.
Immunohistochemistry substantiated previous proteomic studies by confirming the presence of Dystroglycan, Profilin, Clusterin, and Cadherin in the human embryonic vitreous. The cuurent results suggest that in hyalocytes and hyaloid vessels these proteins may play a role in the intraocular vascular regression that occurs during the 2nd trimester. Indeed, Clusterin and Cadherin are not present early in the 2nd trimester but do appear in the hyaloid vasculature by late 2nd trimester, concurrent with the previously reported 3-fold increase in protein concentrations during this time.
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