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Calcium Metabolism of the Normal Rhesus Monkey CALCIUM METABOLISM OF THE NORMAL RHESUS MONKEY Robert S. Harris, … , Joan R. Moor, Robert L. Wanner J Clin Invest. 1961;40(9):1766-1774. https://doi.org/10.1172/JCI104400. Research Article Find the latest version: https://jci.me/104400/pdf CALCIUM METABOLISM OF THE NORMAL RHESUS MONKEY* By ROBERT S. HARRIS, JOAN R. MOOR AND ROBERT L. WANNER t (From the Department of Nutrition, Food Science and Technology, Massachusetts Institute of Technology, Cambridge, Mass.) (Submitted for publication November 14, 1960; accepted May 29, 1961) In research related to human nutrition, it has ratios ranged between 3:1 and 1: 3 (3-6). In become a tradition first to conduct extensive stud- spite of these anomalies, the "endogenous" ex- ies on one or more species of experimental animal, cretion of calcium by normal human beings and and then to attempt to confirm the findings from normal monkeys appears to be similar, and quite these studies on relatively small groups of human unlike that by the dog and the rat. beings. It is important that the animal species The data from studies of the calcium metabolism selected for study should closely resemble the hu- of 29 normal rhesus monkeys (Macaca mulatta) man being with respect to the absorption, metabo- are being reported in this communication, and lism, degradation, and excretion of the nutrient compared with the data from similar studies on under study. The species acceptable for the study human beings conducted by this (2, 7, 8) and by of one nutrient is not necessarily the species of other (3-6, 9-16) laboratories. choice for the study of a different nutrient. The rat and the dog have been used extensively EXPERIMENTAL PROCEDURES in nutrition studies. These species appear to me- All monkeys used in this research were received be- tabolize many nutrients in a manner similar to tween 1955 and 1959 from Okatie Farms,' N. C. On ar- that of human beings, but Wanner and co-work- rival they were placed in individual monkey cages (27 X ers (1) presented preliminary data indicating that 27 X 30 inches) in an air-conditioned (78° F, 60 per cent neither of these species is a satisfactory substitute relative humidity) laboratory designed for metabolic for human beings in studies of calcium metabo- studies with radioactive isotopes. The body weights ranged between 1.8 and 4.9 kg, and all but 3 monkeys lism. They injected trace amounts of Ca45 in- were male. The ages of these captive monkeys were not travenously, and observed that the ratio of Ca45 known. Van Wagenen and Catchpole (17) have sug- excretion in the urine and feces during the next gested that a monkey's age may be calculated from his 5 days was 1: 22 in the rat, 1: 10 in the dog, 2: 1 body weight. On this basis these monkeys ranged be- in the monkey, and 2: 1 in the human being. It tween 9 and 34 months, with 35 per cent of them just under 2 years old. has since been reported by Bronner, Harris, During a short period of acclimatization and ob- Maletskos and Benda (2) that the urine: feces servation, the monkeys were maintained on a nutritionally excretion ratios of six young human subjects adequate diet 2 and distilled water ad libitum. All monk- ranged from 0.50: 1 to 3.2: 1 and averaged 1.9: 1. eys were fasted overnight before each experiment. Each To our knowledge there are no other reports of was then given Ca45Cl, by intravenous injection, by stom- ach tube with 200 or 400 mg calcium (as CaCl2) in similar studies on physiologically normal human aqueous solution, or mixed in the diet after it had been subjects. Data obtained on patients with carci- noma, multiple myeloma, and thyroid disorders 1 Maintained by The National Foundation for the indicate that the excretion of Ca45 is disturbed in quarantine of imported monkeys until cleared of para- these situations, for the urine: feces excretion sites and infectious diseases. 2 Composition of the maintenance diet (g/100 g): 44.0 *A portion of this paper was presented at the Fifth farina (unenriched) ; 22.1 casein (Labco) ; 16.5 sucrose; International Congress on Nutrition, Washington, D. C., 10.0 corn oil (Mazola); 4.0 salt mixture i.v. (18); 3.0 September 1, 1960. This study was aided by grants Celluflour; 0.1 p-aminobenzoic acid; 0.1 inositol; 0.2 from The National Foundation, New York, N. Y. Con- choline chloride; 0.6 ml multivitamin (Vipenta, Roche) tribution no. 431 from the Department of Nutrition, Food drops on alternate days (3 mg thiamine; 3 mg riboflavin; Science and Technology, Massachusetts Institute of 20 mg niacinamide; 1 mg pyridoxine; 3 mg d-pan- Technology, Cambridge, Mass. tothenol; 75 mg ascorbic acid; 1 mg d,l-a-tocopherol t Present address: Animal Nutrition Laboratory, acetate; 2 4Ag vitamin B,2; 5,000 USP U vitamin A; Cornell University, Ithaca, N. Y. 1,000 USP U vitamin D). 1766 CALCIUM METABOLISM IN THE MONKEY 1767 triturated with 120 to 200 nmg calcium as the chloride. of gastrocnemius and biceps muscles were taken from This labeled meal was offered for 30 minutes and the each monkey for analysis. The dissected long bones uneaten residue was analyzed for Ca" and total Ca con- were roentgenographed. tent. Samples of blood, urine, and feces were collected Samples were analyzed for Ca45, total Ca, moisture, at appropriate intervals, and analyzed for Ca" and total and ash content by the methods outlined by Bronner, and Ca content. After the dose, blood samples were generally co-workers (8, 19). The data were statistically analyzed, collected after 0.25, 2, 4, and 6 hours; 1, 2, 3, 5, 7 days; when possible, by methods explained in Snedecor (20); and then weekly. Urine and feces samples were col- SD in this paper refers to "standard deviation." lected daily during the first 7 days, and then 48-hour The monkeys usually defecated in the corners of the samples were collected at weekly or biweekly intervals cages, and the stools were sufficiently firm to permit sep- until the end of the experimental period. arate collection with little opportunity for the leaching Twenty-five of the 29 monkeys used in this research of calcium into the urine. The urine was filtered and were used in subsequent studies of the effects of polio- rinsed through coarse filter paper to remove food myelitis paralysis upon Ca" metabolism. Thus, only 4 particles. of the monkeys may be considered as normal control animals at the time of sacrifice. Three monkeys were RESULTS AND DISCUSSION used in 3 successive experiments in which each received 1. Blood serum data. The total serum calcium the Ca" by intravenous injection, by stomach tube, and in a labeled meal. Several other monkeys were used in content of the 29 monkeys averaged 10.4 mg per a second experiment after sufficient time had elapsed to 100 ml. The specific activities 3 of the serum cal- permit the Ca" levels in the tissues to stabilize. At autopsy the gall bladder, heart, liver, kidneys, bone 3 Specific activity = % administered Ca" per mg total (radius, ulna, humerus, tibia, fibula, femur), and portions calcium in tissue or fluid. 1.0 o 0 8 8 &~~~~~0~~~~ 6 8 8\ 50.l- § ~~~~~~~~TIME, HOURS LI1 '0.01 9 0.01 4- (0 u CQe\U Q18 0 U S1 aL 0fo C2 0ff o s Serum (o) 0 al Al _I or | I ^ | In a - lb Urine (a:) A Ai AA& AA&A O.0 001001~ ~~~~~~~~~~Feces() 0 20 40 60 80 )10 IZ Time, Days FIG. 1. SPECIFIC ACTIVITIES OF SERUM, URINE AND FECES OF NORMAL MONKEYS AFTER INTRAVENOUS INJECTION OF CA5CL2,. 1768 ROBERT S. HARRIS, JOAN R. MOOR AND ROBERT L. WANNER 0 2 4 6 8 10 12 Time, Days FIG. 2. SPECIFIC ACTIVITIES OF SERUM, URINE AND FECES OF NORMAL MONKEYS AFTER STOMACH TUBE FEEDING OF CA CL2. cium samples reached maxima immediately after human adults (5) a maximal specific activity of intravenous injection, approximately 60 minutes 0.12 was reached 15 minutes after injection, and after stomach feeding, and 200 minutes after die- a maximum of 0.04 was reached about 240 min- tary feeding of the Ca45 (Figures 1-3). In each utes after the ingestion of a labeled meal. Bauer, case, the activities dropped at decreasing rates Carlsson and Lindquist (9) have suggested that during subsequent hours. the calcium of the body is contained in two frac- The observation that less Ca45 was absorbed tions: a large relatively inert fraction (99 per from the intestinal tract when it was fed in the cent) contained in the skeletal tissues, and a small diet than when given in aqueous solution by amount (1 per cent) of readily exchangeable cal- stomach tube conforms with the observation by cium. The serum specific activity data in Fig- Sharpe, Peacock, Cooke and Harris (21) that ures 1-3 were used to calculate the exchangeable food solids interfere with the uptake of radioiron calcium and the rate of accretion of calcium per in children. Also, these data are in close agree- day by 24 monkeys by using the method proposed ment with data from similar studies in children, by Bauer and co-workers. The exchangeable cal- in which the specific activities reached a maximum cium, E, was found to be 1.19 g (SD 0.66) or 397 of 0.6 within 15 minutes of intravenous injection, mg per kg body weight in these monkeys. Bauer and a maximum of only 0.2 at 240 minutes after and colleagues reported a value of 0.782 g ex- the ingestion of a labeled meal (7).
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