INTRODUCTION
In humans, genetic deficiency of the enzyme hypoxanthine
phosphoribosyl transferase (HPRT; EC 2.4.2.8), which is
encoded by a gene on the X chromosome, causes Lesch-
Nyhan syndrome (reviewed by Stout and Caskey, 1989).
Lesch-Nyhan patients show a characteristic group of behav-
ioural abnormalities, including spastic cerebral palsy,
choreoathetosis and compulsive selfmutilation. It has been
proposed that at least some of these abnormalities result
from a deficiency in the dopamine-mediated pathways of the
basal ganglia of the brain, since 70-90% depletion of
dopamine was seen in all three Lesch-Nyhan patients
autopsied by Lloyd et al. (1981) and reduced dopamine
metabolite levels have been reported in the cerebrospinal
fluid of Lesch-Nyhan patients throughout life (Silverstein et
al., 1983). As well as these behavioural abnormalities,
effects on haematopoiesis are also present. In different
studies, megaloblastic anaemia, macrocytic and mega-
loblastic changes, haemolytic anaemia and subnormal pop-
ulations of B lymphocytes have been reported (reviewed by
Stout and Caskey, 1989). In heterozygous female carriers,
where X-chromosome inactivation produces a mixture of
functionally HPRT
+
and HPRT
-
cells, there is strong
selection against the HPRT
-
population in haematopoietic
tissues, to the extent that this population is undetectable in
adult lymphocytes and erythrocytes (Dancis et al., 1968;
McDonald and Kelley, 1972). However, in three young
female heterozygotes (7 to 17 years old), between 5 and 10%
of lymphocytes were HPRT deficient, suggesting that the
extent of selection is age dependent (Albertini and De Mars,
1974).
We and others have produced mice deficient in HPRT by
breeding from germ-line chimaeras generated by blastocyst
injection of HPRT-deficient embryonal stem cells (Hooper
et al., 1987; Kuehn et al., 1987). These mice do not show
the behavioural abnormalities of the human condition
(Finger et al., 1988; Hooper, 1988). Trauma to the ears and
flanks caused by overgrooming has been seen in some old
HPRT-deficient mice, but it remains to be established
whether this is due to the mutation (Williamson et al., 1992).
There is a significant reduction in dopamine levels in the
basal ganglia, but the extent of the reduction is much smaller
than in human patients (Finger et al., 1988; Dunnett et al.,
1989; Williamson et al., 1991). Recent work suggests that
HPRT deficiency has less severe neurological and behav-
ioural consequences in mice than in humans because mice
are more reliant on adenine phosphoribosyl transferase
(APRT) than HPRT for purine salvage (Wu and Melton,
1993).
859
Development 118, 859-863 (1993)
Printed in Great Britain © The Company of Biologists Limited 1993
The basis of a previously observed difference in the level
of contribution of hypoxanthine phosphoribosyltrans-
ferase-deficient cells between the haematopoietic and
non-haematopoietic tissues of chimaeric and heterozy-
gous mice has been clarified by studying two populations
of female mice that differ only in that one is heterozy-
gous for a null allele at the hprt locus and the other is
wild type at this locus. Both populations are heterozy-
gous for an electrophoretic variant allele at the X-linked
Pgk-1 locus, so that X-chromosome inactivation
generates cells expressing different isozymes of phos-
phoglycerate kinase which can be assayed to monitor cell
selection. The results show that hypoxanthine phospho-
ribosyltransferase deficiency itself, rather than an effect
of another X-linked gene, causes a reduced level of con-
tribution to haematopoietic tissues. Further, the extent
of the depletion increases significantly with age, and this
effect is due to a progressive reduction in the level of con-
tribution to haematopoietic tissues rather than to an
increase in the level of contribution to non-haematopoi-
etic tissues.
Key words: haematopoiesis, HPRT, Lesch-Nyhan syndrome
SUMMARY
Age-dependent selection against hypoxanthine phosphoribosyl
transferase-deficient cells in mouse haematopoiesis
K. Samuel
1
, A. R. Clarke
2
, J. D. Ansell
1,3
and M. L. Hooper
2,3,
*
1
Institute of Cell, Animal and Population Biology, Ashworth Laboratories, University of Edinburgh, West Mains Road,
Edinburgh EH9 3JT, UK
2
Cancer Research Campaign Laboratories, Department of Pathology, University of Edinburgh, Teviot Place, Edinburgh EH8
9AG, UK
3
AFRC Centre for Genome Research, King’s Buildings, University of Edinburgh, West Mains Road, Edinburgh EH9 3JQ, UK
*Author for correspondence at address
2