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Heat Shock Protein Concentration and Clarity of Porcine Lenses Incubated at Elevated Temperatures
Author(s) -
Dzialoszynski Tomasz M,
Trevithick John R,
Noble Earl G
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.128
Subject(s) - incubation , lens (geology) , hsp70 , heat shock protein , shock (circulatory) , blot , hsp27 , chemistry , biology , biochemistry , medicine , gene , paleontology
Purpose: to quantify the concentration of heat shock proteins in lenses and culture medium in organ culture at elevated temperatures. Methods: Pig lenses were dissected aseptically and incubated in medium M199 without serum for 4 days and those with protein leakage less than 10 mg/L were taken for heat shock (HS). HS was performed by incubation for 1 hr at temperatures from 37°C to 55°C. After incubation for 24 hr, the lenses were weighed and homogenized and the medium was collected. The medium and lens homogenates were analysed for Hsp 70 and Hsp 27 by western blotting. ScantoxTM and analysis of lens photographs were used to estimate cataract focal length variability. Results: Hsp 70 was generally increased in the lens at temperatures above 39°C but the response was diminished at 55°C. No changes in Hsp27 concentration in the lens were observed. Interestingly, Hsp70 levels also increased in the culture medium after HS at 42.5°C. Increased temperatures were also associated with greater focal length variability and with loss of lens weight during the 24 hours following HS. Conclusions: Hsp70 concentration in the lens, its release, and the focusing ability of the lens were altered in response to HS The observation that Hsp70 is released by the stressed lens may suggest an important paracrine function, given the potential signaling properties of these proteins. Supported by CSA and NSERC 8170‐05 RGPIN