OCCURRENCE OF FREE AMINO ACIDS — VERTEBRATES 461 
Invited Discussion 
INCREASE IN URINARY AMINO ACIDS ASSOCIATED 
WITH PANTOTHENIC ACID DEFICIENCY IN THE RAT 
JOY D. MARKS anp HELEN K. BERRY 
Department of Pediatrics, University of Cincinnati; 
The Childven’s Hospital Research Foundation, Cincinnati, Ohio (U.S.A.) 
Chromatographic separation and identification of the amino acid constituents of 
urine indicate that a rather specific “pattern” of urinary amino acids may be associ- 
ated with age, physiological condition and hereditary metabolic disease. Information 
about the metabolic state of the subject is provided by qualitative and quantitative 
data regarding the urinary amino acids. 
The occasional finding of high levels of alanine in the blood and urine of children 
suggested the possibility of pantothenate deficiency impairing the coenzyme A 
metabolism of the child. The experiments described below were designed to test the 
hypothesis that alanine would be excreted by pantothenate-deficient rats. In the 
course of experiments aimed at producing congenital malformations in rats, pregnant 
rats fed pantothenic acid-deficient diet were also available. These rats were used as 
a means of investigating the amino acid composition of the urine during normal 
pregnancy and of studying the effect of pantothenate deficiency on the composition 
of the urine during the gestational period. The results of these investigations repre- 
sent an example of a biochemical lesion that can be experimentally produced. 
METHOD 
Sprague-Dawley albino rats, of commercial stock, were placed in metabolic cages 
and maintained on the pantothenic acid-deficient diet (Nutritional Biochemicals, 
Cleveland). Urine was collected in graduated cylinders under toluene for 24-h 
periods or longer. Measured aliquots (usually 1/500 of the total daily volume) were 
chromatographed according to standard procedures! and total amino acid excretion 
(mg/day/rat) was determined. Pantothenic acid deficiency was produced by a method 
similar to that described by NELSON et al.?. Oral doses of w-methylpantothenate 
(1-2 mg) were administered to assure the production of deficiency. With the pregnant 
rats several feeding schedules were tried to establish a state of pantothenic acid 
deficiency, but none of them was severe enough to cause resorption or premature 
termination of pregnancy. 
RESULTS 
Pantothenic acid deficiency in male weanling rats. Two male weanling rats were fed 
a complete diet and two were given a pantothenic acid-deficient diet plus w-methyl- 
pantothenate for a 3-week period. The rats fed a complete diet excreted glutamic 
acid, glycine, and alanine (approx. 0.8—1.2 mg/day of each). Only traces of lysine, 
glutamine, threonine and serine were excreted (0.2 mg/day). The rats fed the deficient 
diet lost their hair and became weak and feeble by 3 weeks. Alanine excretion in- 
References p. 464 
