Purchase this article with an account.
N Tsuji, F Zhang, AL McNeal, JD Drake, DG Hwang; Detection of Leukocyte Chemotaxis Inhibitor slit2 mRNA in Human Corneas . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1619.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate the existence of slit2, an inhibitory factor for chemokine-induced leukocyte chemotaxis, in human corneas. Methods: Donor corneal rims and primary cultured corneal epithelial, stromal and endothelial cells were used for this experiment. Slit2 mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in primary cultured cells and further confirmed by digoxigenin-labeled in situ hybridization (DIG-ISH) in corneal tissue. (1) RT-PCR: Total RNAs were harvested from confluent primary cultured corneal epithelial (n=8), stromal (n=2) and endothelial cells (n=15). RT-PCR was performed for slit2. (2) DIG-ISH: The PCR product of slit2 was cloned and sense/anti-sense DIG labeled RNA probes were made. Corneal rims (n=2) were fixed in 4% paraformaldehyde followed by 30% sucrose cryoprotective treatment and frozen sections were made on silylated glass slides. The hybridization was visualized by colorimetric detection system. Results: Slit2 mRNA was found in all corneal epithelial, stromal and endothelial cells of human donor corneas by both RT-PCR and anti-sense DIG-ISH. Conclusion: This is the first report of slit2 in human corneas. This may provide additional clues to understanding the mechanisms underlying the immune privilege status of the eye.
This PDF is available to Subscribers Only