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  • Title: Isolation of mutant renal (LLC-PK1) epithelia defective in basolateral, Na(+)-independent glucose transport.
    Author: Mullin JM, Snock KV, McGinn MT, Kofeldt LM.
    Journal: Am J Physiol; 1989 Dec; 257(6 Pt 2):F1039-49. PubMed ID: 2603953.
    Abstract:
    To obtain mutant renal epithelia defective in the uptake of 2-deoxyglucose (Na(+)-independent glucose transport), LLC-PK1 renal epithelial cells were subjected to a combination of DNA alkylation, tritium-suicide with 2-[3H]deoxyglucose, and replica plating. One of the mutant sublines obtained, LLC-PK1M-7A, possesses only 40% of the 2-deoxyglucose uptake rate of the parent line, LLC-PK1M, and this defect is stable over at least 30 population doublings. Initial rate of 3-O-methylglucose transport into these mutant cells is only 20% of the parental rate, and efflux of 3-O-methylglucose from the mutant cells is correspondingly low. Glucose metabolism in the mutant cells does not appear to be altered, nor is free glucose accumulated in the cells against a concentration gradient. The uptake rate of L-leucine is the same in both mutant and parent, whereas the (Na(+)-dependent) uptake of alpha-methyl-D-glucoside and methylaminoisobutyric acid is greater in the mutant than the parent. The population doubling times of LLC-PK1M and LLC-PK1M-7A cultures are similar. LLC-PK1M-7A cell morphology is similar to LLC-PK1M when cultures are subconfluent, but on reaching confluence, LLC-PK1M-7A appear larger than LLC-PK1M cells. Their measured cell volume is then 150% of the volume of the parent cells. Future studies of the LLC-PK1M-7A mutant, and acquisition of additional mutant sublines, should elucidate the roles of transepithelial glucose transport in proximal tubular cell physiology.
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