Journal List > Korean J Ophthalmol > v.19(2) > 1024400

Kim, Yu, Jeong, Kim, and Kim: Protective Effect of Heat Shock Proteins 70.1 and 70.3 on Retinal Photic Injury after Systemic Hyperthermia

Abstract

Purpose

This study aimed to determine the relationship between the heat shock protein 70 from hsps70.1 and 70.3 on retinal photic injury after systemic hyperthermia.

Methods

Eight-week-old female C57BL/6 mice were kept at a constant temperature of 41~42℃ for 25~30 minutes. After dark-adaptation for 8 hours, intense light of 11000 lux was maintained for 6 hours. Histology and immunohistochemistry for the inducible heat shock protein 70 (hsp70), the constitutive heat shock protein 70 (hsc70), and western blot analysis, reverse transcriptase-polymerase chain reaction for hsp70.1 and hsp70.3 were performed just before photic injury and after 1, 4, 7, and 14 days.

Results

Light-induced retinal degeneration was prevented by thermotolerance. After hyperthermia, hsp70 was densely expressed in the inner segment of the photoreceptor layer on the photic injury. Hsp70 expression increased for 4 days after photic injury and slowly decreased thereafter. mRNA from hsp70.3 was induced earlier than that of hsp70.1.

Conclusions

Retinal photic injury was prevented by hyperthermia-induced hsp70. Hsp70 from hsp70.3 may be a rapid and short-lived responder, and that from hsp70.1 is a slower and more sustained responder. Hsp70 from hsp70.3 may be an initial retinal chaperone while hsp70 from hsp70.1 may be a sustained chaperone.

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Fig. 1.
Histology of the retina after photic injury with or without hyperthermia. On photic injury without previous hyperthermia, diffuse disarrangements in the ONL and photoreceptor cell layer was observed (white arrows in B). However, photic injury after hyperthermia made no definite change in retina structure (white arrows with dotted line in B’). Hematoxylin & Eosin staining, magnification ×400 (A, A’: before photic injury; B, B’: 14 days after photic injury, L; light stress only, L+H; light stress after systemic hyperthermia)
kjo-19-116f1.tif
Fig. 2.
Immunohistochemistry for hsp70 and hsc70 in the retina. Hsp70 expression was most prominent in the inner segment of photoreceptors (black arrows) for 4 days after photic injury following hyperthermia. Hsc70 expression was detected in most retinal layers (black arrows with dotted line), and nearly constant without temporal or spatial difference. Hematoxylin & Eosin staining, magnification × 400 (A, A’: before photic injury; B, B’: 1 day after photic injury; C, C’: 4 days; D, D’: 7 days; E, E’: 14 days).
kjo-19-116f2.tif
Fig. 3.
Western blot analysis for hsp70. In A, enhanced hsp70 was due to hsp induction by systemic hyperthermia. The expression of hsp70 was sustained for 7 days. Although changes were subtle, the quantitative measurement showed that production of hsp70 increased for 4 days after photic injury and slowly decreased thereafter. (A: before photic injury; B, C, D and E: 1, 4, 7 and 14 days after photic injury, respectively.)
kjo-19-116f3.tif
Fig. 4.
Semiquantitative analysis of mRNA expression, RT-PCR, for hsp70.1 and hsp70.3. mRNA expression of hsp70.1 reached a peak 4 days after light stress and slowly decreased until day 14. The mRNA expression in hsp70.3 was most prominent at day 1 and slowly decreased thereafter. (A, A’: before photic injury; B, B’: 1 day after photic injury; C, C’: 4 days; D, D’: 7 days; E, E’: 14 days)
kjo-19-116f4.tif
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