Normalization of lens protein kinase Cgamma in galactosemic dogs by a novel aldose reductase inhibitor.
The purpose of this study was to determine the effects of a novel aldose reductase inhibitor on lens protein kinase Cgamma (PKCgamma) levels in galactosemic dogs. Six-month old Beagles (12 total; 6 male and 6 female) were made galactosemic by feeding a diet of 40% galactose for 6 weeks. Three dogs per group were fed either control, normal diet, 40% galactose diet, 40% galactose diet with aldose reductase inhibitor at 100 mg/kg body weight per day given orally, or a control diet with aldose reductase inhibitor alone (1-H,7-H-5alpha,6,8,9-tetrahydro-1-oxopyran[4,3-beta](1) benzopyran, referred to herein as HAR-1). Lenses were removed and analyzed for toxicity by pathological examination. Lens polyol concentrations were determined by GC/MS. PKCgamma levels were determined by Western blot and by reverse transcriptase-polymerase chain reaction (RT-PCR). No toxicity was observed from the aldose reductase inhibitor when given at 100 mg/kg body weight per day for 6 weeks. Galactosemic dogs showed deterioration of lens cells. Deterioration included vacuole formation in the lens, cell lysis, and loss of cell nuclei. Galactosemic dogs given the HAR-1 appeared identical to control dogs. Polyol concentrations in the lenses were reduced by 50% in dogs fed the 40% galactose diet with the aldose reductase inhibitor, HAR-1. PKCgamma protein levels were reduced in the galactosemic dog lenses, but synthesis of PKCgamma was not affected, as measured by RT-PCR. The PKCgamma protein levels were similar to controls in dogs given the aldose reductase inhibitor, HAR-1, even when polyol concentrations remained 50% elevated above control levels. HAR-1, when given to control dogs, caused a reduction in the synthesis of PKCgamma mRNA but not in total PKCgamma protein levels. This study demonstrates the use of a novel aldose reductase inhibitor to control changes in PKCgamma in dog lens, a PKC that is known to control gap junction activity.
Takemoto
Dolores J
DJ
Department of Biochemistry, Willard Hall, Kansas State University, Manhattan, KS 66506, USA. Dtak@ksu.edu
Harris
Richard
R
Brightman
Al
A
McGill
John
J
Hua
Duy
D
Davidson
Harriet
H
Fenwick
Brad
B
Wagner
Lynn M
LM
D000085:Acetates / Q000008:administration & dosage / Q000627:therapeutic use*; D000284:Administration, Oral; D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors*; D000818:Animals; D001578:Benzopyrans / Q000008:administration & dosage / Q000627:therapeutic use*; D015153:Blotting, Western / Q000662:veterinary; D017931:DNA Primers; D004283:Dog Diseases / Q000188:drug therapy* / Q000201:enzymology* / Q000473:pathology; D004285:Dogs; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000627:therapeutic use*; D005260:Female; D005693:Galactosemias / Q000188:drug therapy / Q000201:enzymology / Q000662:veterinary*; D007908:Lens, Crystalline / Q000201:enzymology*; D008297:Male; D011493:Protein Kinase C / Q000032:analysis* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
C433083:1H,7H-5a,6,8,9-tetrahydro-1-oxopyrano(4,3-b)(1)benzopyran-3-acetic acid; D000085:Acetates; D001578:Benzopyrans; D017931:DNA Primers; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D000449:Aldehyde Reductase; C087857:protein kinase C gamma; D011493:Protein Kinase C