McDowell Colleen M, Dutca Laura M, Thompson Stewart, Riker Megan, Hedberg-Buenz Adam, Meyer Kacie J, Anderson Michael G
Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA, USA.
Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA, USA; Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA; Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA.
Exp Eye Res. 2025 Mar;252:110266. doi: 10.1016/j.exer.2025.110266. Epub 2025 Jan 31.
Intraocular pressure (IOP) follows a circadian rhythm. In both humans and mice, IOP is normally slightly elevated at night during the dark phase of the light cycle. In studying a strain of mice for possible indices of glaucoma, we incidentally discovered that C57BL/6J mice homozygous for the beige-J mutation of the Lyst gene lack a circadian fluctuation in IOP. Instead of having an elevated dark phase IOP, homozygotes exhibit a uniform IOP characteristic for light period values of C57BL/6J mice. The beige-J mutation results from deletion of a single isoleucine amino acid in the LYST WD40 motif likely to influence protein-protein interactions. Based on the literature, we hypothesized that CSNK2B (casein kinase 2, beta polypeptide) might be a relevant interacting protein, which we confirmed with a pulldown assay as a binding partner of wild-type, but not beige-J encoding, LYST protein. Treating wild-type mice with 4,5,6,7-tetrabromobenzotriazole (TBB), a casein kinase 2 inhibitor, recapitulated the beige-J mutant phenotype in preventing a rise in IOP during the dark period. Together, these results identify Lyst beige-J mice as a new strain for studying circadian IOP regulation and point to casein kinase 2 as a key molecule of interest.
眼压(IOP)遵循昼夜节律。在人类和小鼠中,眼压通常在光周期的黑暗阶段夜间略有升高。在研究一种可能作为青光眼指标的小鼠品系时,我们偶然发现,Lyst基因米色-J突变的纯合C57BL/6J小鼠缺乏眼压的昼夜波动。纯合子没有夜间眼压升高,而是表现出与C57BL/6J小鼠光照期值一致的眼压特征。米色-J突变是由于LYST WD40基序中一个异亮氨酸氨基酸缺失所致,这可能会影响蛋白质-蛋白质相互作用。基于文献,我们假设CSNK2B(酪蛋白激酶2,β多肽)可能是一种相关的相互作用蛋白,我们通过下拉试验证实它是野生型LYST蛋白(而非编码米色-J的LYST蛋白)的结合伴侣。用酪蛋白激酶2抑制剂4,5,6,7-四溴苯并三唑(TBB)处理野生型小鼠,重现了米色-J突变体表型,即在黑暗期阻止眼压升高。总之,这些结果确定Lyst米色-J小鼠是研究昼夜眼压调节的新菌株,并指出酪蛋白激酶2是一个关键的相关分子。