Targeted Deletion of Both Thymidine Phosphorylase and Uridine Phosphorylase and Consequent Disorders in Mice

Abstract

Thymidine phosphorylase (TP) regulates intracellular and plasma thymidine levels. TP deficiency is hypothesized to (i) increase levels of thymidine in plasma, (ii) lead to mitochondrial DNA alterations, and (iii) cause mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). In order to elucidate the physiological roles of TP, we generated mice deficient in the TP gene. Although TP activity in the liver was inhibited in these mice, it was fully maintained in the small intestine. Murine uridine phosphorylase (UP), unlike human UP, cleaves thymidine, as well as uridine. We therefore generated TP-UP double-knockout (TP^−/− UP^−/−) mice. TP activities were inhibited in TP^−/− UP^−/− mice, and the level of thymidine in the plasma of TP^−/− UP^−/− mice was higher than for TP^−/− mice. Unexpectedly, we could not observe alterations of mitochondrial DNA or pathological changes in the muscles of the TP^−/− UP^−/− mice, even when these mice were fed thymidine for 7 months. However, we did find hyperintense lesions on magnetic resonance T₂ maps in the brain and axonal edema by electron microscopic study of the brain in TP^−/− UP^−/− mice. These findings suggested that the inhibition of TP activity caused the elevation of pyrimidine levels in plasma and consequent axonal swelling in the brains of mice. Since lesions in the brain do not appear to be due to mitochondrial alterations and pathological changes in the muscle were not found, this model will provide further insights into the causes of MNGIE.

We gratefully acknowledge R. Wesselshmidt (Genomesystems, Inc.) for culture of the ES cells and production of the chimeric mice. We also thank E. Sudo for the mice breeding and the PCR analyses; N. Hirata, T. Kodama, and N. Misawa for histological analysis; and G. Yamada, A. Aiba, M. Nakagawa, and H. Yamamoto for insightful comments. We thank S. Narisawa for many helpful suggestions.

This work was supported in part by a grant-in-aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.