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					PROCEEDINGS OF THE BIOCHEMICAL SOCIETY                                                                          21 p

are apparently correlated with changes in dihydroxy-       of sucrose-fed animals were always higher than those
acetone phosphate concentration. In non-diabetic           of controls. ATP citrate lyase (EC and 'malic'
rats they are both decreased in starvation and in-         enzyme (EC increased in activity after
creased 48 h after re-feeding with glucose, glycerol or    weaning to a maximum at 35-40 days of age and then
sucrose. Further, in diabetes the activity of liver-type   fell to the end of the experiment, and feeding with
pyruvate kinase and dihydroxyacetone phosphate             sucrose caused elevated activities at all ages studied.
concentration are decreased, and in this state both        Extramitochondrial aconitate hydratase (EC
are raised by treatment with insulin and by re-feeding     and NADP-dependent isocitrate dehydrogenase (EC
with sucrose or glycerol but not with glucose. How- activities fell after weaning until 25-30 days
ever, no causal relationship has been established.         of age and then rose throughout the rest of the
   The authors are indebted to the Science Research
                                                           experimental period. From 30 days of age the activi-
Council for a grant to J. M. G.                            ties of both these enzymes were lowered by feeding
                                                           with sucrose. Glucose 6-phosphate dehydrogenase
Bailey, E., Taylor, C. B. & Bartley, W. (1968) Nature      (EC activity rose rapidly after weaning to
   (London) 217,471                                        40 days of age and then remained constant for the
Sillero, A., Sillero, M. A. G. & Sols, A. (1969) Eur. J.   remainder of the experimental period. Feeding with
   Biochem. 10, 351                                        sucrose increased this enzyme activity at all ages
Weber, G., Singhal, R. L., Stamm, N. B., Lea, M. A. &      studied.
   Fisher, E. A. (1966a) Advan. Enzyme Regul. 4, 59
Weber, G., Lea, M. A., Fisher, E. A. & Stamm, N. B.           We thank the Medical Research Council for a grant to
   (1966b) Enzymol. Biol. C/in. 7, 11                      J. R. W.
The Effect on the Activities of some Hepatic               Bailey, E., Taylor, C. B. & Bartley, W. (1968) Nature
                                                              (London) 217, 471-472
Enzymes of Weaning Rats on to a Diet High in               Lockwood, E. A., Bailey, E. & Taylor, C. B. (1970)
Sucrose Content                                               Biochem. J. 118,155-162
                                                           Taylor, C. B., Bailey, E. & Bartley, W. (1967) Biochem. J.
By JENNIFER R. WEBB and E. BAILEY (Department of              105, 717-722
Biochemistry, University of Sheffield, Sheffield
S10 2TN, U.K.)
                                                           Changes in the Activities of some Kidney
   There is a marked increase in hepatic lipogenesis       Enzymes during Development of the Rat
and the activities of enzymes associated with lipo-
genesis on weaning rats from the high-fat-content          By CHRIsTINE A. HAUSER, ELIZABErH A. LOCKWOOD
milk diet of suckling to the high-carbohydrate             and E. BAILEY (Department of Biochemistry, Univer-
(starch)-content laboratory diet (Taylor et al., 1967;     sity of Sheffield, Sheffield S10 2TN, U.K.)
Lockwood et al., 1970), Hepatic lipogenesis is also
considerably higher in rats fed on sucrose rather than        Although changes in the activities of many hepatic
on starch-containing diets (Bailey et al., 1968). To       enzymes during development of the rat are well docu-
investigate further the involvement of diet in the post-   mented, little is known about the developmental
weaning increase in lipogenesis we have studied the        changes that occur in the activity of rat kidney en-
effect on the activity of some enzymes involved in         zymes (for review see Greengard, 1971). We have
carbohydrate conversion into fatty acids of weaning        measured the postnatal changes that occur in the
female rats on to a diet high in sucrose content.          activities of the following kidney enzymes: hexo-
   Fructokinase (EC activity increased after      kinase (EC, pyruvate kinase (EC,
weaning (at 21 days of age) to a maximum at 40-45          'malic' enzyme (EC, glucose 6-phosphate
days and then fell to pre-weaning values by the end        dehydrogenase (EC, extramitochondrial
of the experiment (70 days of age), and feeding with       NADP-dependent isocitrate dehydrogenase (EC
sucrose had little effect on the enzyme activity., extramitochondrial aconitate hydratase
Phosphofructokinase (EC and fructose 1,6-        (EC, ATP citrate lyase (EC and
diphosphatase (EC activities varied little       fructose 1,6-diphosphatase (EC We have
during the period studied and dietary sucrose had          also measured developmental changes in the in-
little effect on the activity of either enzyme. Aldolase   corporation of [U-14C]acetate into lipid by kidney
(EC activity was constant between 21 days        homogenates. The experiments were carried out with
and 40 days of age, rose to a maximum at 50 days           both male and female rats weaned at 21 days of age,
and then fell to 70 days of age. Feeding with sucrose      and unless otherwise stated similar results were
caused slightly elevated aldolase activities at all ages   obtained with both sexes.
studied. Pyruvate kinase (EC activity rose          Hexokinase activity increases after birth to a maxi-
steadily during the experiment to a maximum at             mum at 30 days of age, thereafter falling to reach
about 50-60 days of age, and activities in the livers      adult values by 50 days of age. These results extend
22 p                                                   PROCEEDINGS OF THE BIOCHEMICAL SOCIETY

the findings of Sydow (1969). The activity of pyruvate        Male Wistar rats were force-fed on a protein-free
kinase, the other glycolytic enzyme studied, changed       purified diet for 5-7 days. In agreement with our
very little during the period investigated. Glucose        previous work, the activity of microsomal UDP-
6-phosphate dehydrogenase activity fell slightly after     glucuronyltransferase in livers from these animals
birth to a minimum at 20 days of age and then rose         was substantially higher than in those from control
gradually to reach adult values at 50 days of age.         animals fed on the same diet supplemented with 18%
Isocitrate dehydrogenase and 'malic' enzyme ex-            casein (protein-deficient, 148.7±13.4nmol of glucur-
hibited similar developmental patterns, the activities     onide formed/h per mg of microsomal protein;
of both enzymes rising steadily throughout postnatal       controls, 63.2±6.8).
development. The activity of kidney 'malic' enzyme            Although the weights of microsomal protein and
was 50 % higher in female rats than in male rats.          phospholipid per g of liver were both 30 % lower in
   Extramitochondrial aconitate hydratase activity         protein-deficient animals than in control animals,
increased after birth, reaching adult values by 20 days    there was no significant difference in the ratio of
of age. ATP citrate lyase activity, which was very low     phospholipid to protein between the groups (protein-
throughout development, fell after birth to a mini-        deficient, 1.07±0.03g/g; controls, 1.00±0.01 g/g).
mum at 20 days of age and then rose steadily to adult         The microsomal phospholipid compositions were
values. [U-14C]Acetate incorporation into lipid rose       similar in the two groups, but there were significant
after birth to a maximum at 20 days of age and then        differences in the content of phosphatidylinositol
fell to a minimum at 50 days of age, thereafter rising     (protein-deficient, 6.1±0.5% of total phospholipid;
to adult values. Fructose 1,6-diphosphatase activity       controls 8.7±0.3%.) and sphingomyelin (protein-
rose after birth, reaching adult values at about 30 days   deficient, 7.0 ±0.6 %; controls, 4.2 ±0.3 Y.). Moreover,
of age, thus exhibiting a similar developmental pattern    lysophosphatidylcholine (7.4±0.5%) and lysophos-
to that of glucose 6-phosphatase and gluconeo-             phatidylethanolamine (3.1 ±0.9%) were found with
genesis reported by Zorzoli et al. (1969).                 protein-deficient rats whereas no lysophosphatides
                                                           were detected with control animals.
  We thank the Medical Research Council for grants to         Hanninen & Puukka (1971) report that addition of
C. A. H. and E. A. L.                                      lysophosphatides to rat liver microsomal fractions or
Greengard, 0. (1971) Essays Biochem. 7, 159-205            their treatment with phospholipase A increases
Sydow, G. (1969) Hoppe-Seyler's Z. Physiol. Chem. 350,     UDP-glucuronyltransferase activity. Treatment of
  263-268                                                  our control microsomal preparations with phospho-
Zorzoli, A., Turkenkopf, I. J. & Mueller, V. L. (1969)     lipase A showed that a degree of phospholipid hydro-
  Biochem. J. 111, 181-185                                 lysis similar to that observed in preparations from
                                                           protein-deficient rats increased enzyme activity 2-3-
Effects of Protein Deficiency on Uridine                      We therefore suggest that the elevated enzyme
Diphosphate Glucuronyltransferase Activity and             activity in microsomal fractions from protein-defici-
Phospholipid Composition of Rat Liver                      ent rats is probably due to the presence of lysophos-
Microsomal Fraction                                        phatides. The involvement of other factors such as
                                                           the alterations of phosphatidylinositol and sphingo-
By ALLAN B. GRAHAM, GEOFFREY C. WOOD and                   myelin contents cannot, however, be excluded.
BARRY G. WOODCOCK (Drug Metabolism Research
Unit, Department of Pharmaceutical Chemistry,                We gratefully acknowledge support for this work from
University ofStrathclyde, Glasgow G1 1 XW, U.K.)           the Medical Research Council and the Science Research
   Protein deficiency markedly increases the activity
of rat liver microsomal UDP-glucuronyltransferase          Attwood, D., Graham, A. B. & Wood, G. C. (1971)
with o-aminophenol or p-nitrophenol as acceptor              Biochem. J. 123, 875-882
(Wood & Woodcock, 1970; Woodcock & Wood,                   Graham, A. B. & Wood, G. C. (1969) Biochem. Biophys.
                                                             Res. Commun. 37, 567-575
1971). Since the enzyme's activity is affected by treat-   Hanninen, 0. & Puukka, R. (1971) Chem.-Biol. Inter-
ment of microsomal preparations with agents per-             act. 3, 282-284
turbing the phospholipids of the microsomal mem-           Wood, G. C. & Woodcock, B. G. (1970) J.Pharm.Pharma-
brane (Graham & Wood, 1969; Attwood et al.,                  col. 22, 60S-63S
1971; Hanninen & Puukka, 1971), it was suggested           Woodcock, B. G. & Wood, G. C. (1971) Biochem.
(Woodcock & Wood, 1971) that protein deficiency              Pharmacol. 20, 2703-2713
might affect enzyme activity by altering the structure
of the microsomal membrane. We have therefore
determined the phospholipid composition of liver
microsomal preparations from normal and protein-
deficient rats.