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. 2019 Dec:189:107811.
doi: 10.1016/j.exer.2019.107811. Epub 2019 Sep 21.

Significant upregulation of small heat shock protein αA-crystallin in retinal detachment

Sumaya Hamadmad  1 , Mohd Hussain Shah  1 , Rania Kusibati  1 , Bongsu Kim  1 , Brandon Erickson  1 , Tyler Heisler-Taylor  1 , Sanjoy K Bhattacharya  2 , Mohamed H Abdel-Rahman  3 ; OVER-PVR Study Group; Colleen M Cebulla  4
Collaborators, Affiliations

Significant upregulation of small heat shock protein αA-crystallin in retinal detachment

Sumaya Hamadmad et al. Exp Eye Res. 2019 Dec.
No abstract available

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Figures

Fig. 1.
Fig. 1.
Western Blot analysis of αΑ-crystallin in murine retinal detachment eyes. Retinal detachment (RD) was introduced in murine left eyes as described in methods section, right eyes were used as control (C). Retinas were harvested 1 week (n = 5) and 2–5 weeks (n = 6) after RD and equal amount of total protein was loaded for electrophoresis and subsequent Western Blotting with anti αΑ-crystallin antibody. Western blots representative of at least 3 different experiments are shown in A. The expression of α-tubulin was used as a control. Quantification of at least 3 different experiments is described in B.
Fig. 2.
Fig. 2.
Immunohistochemistry of αΑ-crystallin in murine retinal detachment eyes. Retinal detachment (RD) was introduced in murine left eyes and right eyes were used as control (C). Eyes were harvested 1 week after RD, fixed and sectioned as described in methods section. Sections were stained with αA-crystallin antibody (green) and counterstained with DAPI (blue) for nuclei. An increased diffuse αA-crystallin reactivity was observed diffusely in RD retina as compared to control (C). ONL: outer nuclear layer; INL: inner nuclear layer; GCL: ganglion cell layer.
Fig. 3.
Fig. 3.
Quantification of αΑ-crystallin mRNA using RT-PCR. RNA was collected from retinal samples 3 days, 1 week, or 2 weeks post RD (n = 4/group). RT-PCR shows a time dependent increase in αA-crystallin at the transcription level starting 3 days post RD as compared to control eyes. Error bars represent SEM of biological replicates.
Fig. 4.
Fig. 4.
αA-crystallin measurement in control and RD patient vitreous samples using ELISA. (A) Vitreous samples were collected from RD patients (n = 17) and non RD controls (n = 20). The samples were selectively drawn from pseudophakic individuals to rule out the lens as a source of αA-crystallin. Colorimetric competitive ELISA was used to measure αA-crystallin level in the vitreous. αA-crystallin levels were significantly increased in the vitreous of RD patients compared to their control counterparts (P-value = 0.038). (B) αA-crystallin levels were evaluated in the RD patients (n = 17) divided into smaller vs larger RDs (1 quadrant involved vs. > 1 quadrant) (p = 0.27).
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References

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