ADRENAL ASCORBIC ACID CHANGES IN THE GONADECTOMIZED GOLDEN HAMSTER FOLLOWING A SINGLE INJECTION OF DIETHYLSTILBESTROL

BERNARD C. WEXLER Department of Anatomy, Stanford University, School of Medicine, Stanford, California

C

HRONIC treatment of golden hamsters with diethylstilbestrol (Kirkman and Bacon, 1950), with estradiol (Kirkman, 1951) is followed by the appearance of malignant renal tumors in males, while females tend to show kidney damage similar to that found in glomerulonephritis (Matthews, Kirkman and Bacon, 1947; Kirkman and Bacon, 1950). It is possible that one or both of these pathological conditions may follow a stilbestrol-induced disturbance of the normal pituitary-adrenal relationship. Using adrenal ascorbic acid alterations as evidence for ACTH-release (Sayers, 1950), it has been found that following a single injection of stilbestrol there is a marked decrease of vitamin C in the female hamster but only a moderate decrease in the male (Wexler, 1951). In order to study this marked sex difference in adrenal response, the above experiment (Wexler, 1951) was repeated using gonadectomized animals. METHODS Hamsters (Cricetus auratus) were gonadectomized at 50 days of age, and at 90 days were divided into two separate groups according to sex. Each animal was given an intraperitoneal injection of 0.06 rag. of diethylstilbestrol (Lilly) in \ cc. of cottonseed oil and sacrificed by decapitation 0, \, 1, 3, 6, 12, 24, 36, 48 and 96 hours later. At the time of sacrifice the adrenal glands were removed as quickly as possible, trimmed free of fat, weighed to the nearest 0.01 mg. on a Roller-Smith torsion balance, homogenized, and analyzed for their ascorbic acid content according to the method of Bessey (1938). Each homogenate contained a pool of one adrenal from each of two animals. RESULTS

Quantitative chemical analysis for adrenal ascorbic acid in the golden hamster showed marked increase following gonadectomy (cf. Tables 1, 2 and Figures 1, 2). The adrenal ascorbic acid content in castrated and spayed hamsters was twice that found in intact males and females. The adrenal ascorbic acid content of the spayed hamster underwent Received for publication July 5, 1951. Present address: Baxter Laboratories, Morton Grove, Illinois. 531

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WEXLER

532

Volume 50

TABLE 1. CHANGES IN ADRENAL ASCORBIC ACID FROM SPAYED AND NORMAL FEMALE HAMSTERS AT VARIOUS INTERVALS FOLLOWING A SINGLE INJECTION OF 0.06 MG. OF DIETHYLSTILBESTROL

Spayed Time (hours)

No. of animals

Average adrenal weight (mgs.)

Average body weight (Gms.)

0

16 8

6.4 6.9 5.4

129 126 121 117 112 112 118 111 109

115

i

1

3 6 12 24

8 8 8 8 8 8

5.1

48

8

5.4 5.9 4.9 5.0 3.7

96

8

4.5

36

Average mgs. ascorbic acid per 100 gms. adrenal wt. with S.E.M. 253±23.9 160±16.5 184 + 10.2 209 ±16.9 251± 9.9 243±26.1 237+ 7.3 243 ±32.9 256 ±28.9 252 + 17.0

Normal .0014 .0080 .1136 Normal Normal Normal Normal Normal Normal

115 ± 2.1 92± 3.8 71+ 1.2 95± 3.7 165±17.4 132+10.4 124±16.3 114± 3.1 114+ 4.8

Normal .0001 .0001 .0001 .0004 .1096 .5823 Normal Normal

V

value

Normal 0 * 1

3 6 12 24

36 48

16

8.9 8.4 9.3

8 8 8 8

11.0 10.3

8 16

8

8

.

8.5 9.5 8.7 8.5

125 110 125 130 110 117 114 124 144

TABLE 2. CHANGES IN ADRENAL ASCORBIC ACID FROM CASTRATE AND NORMAL MALE HAMSTERS AT VARIOUS TIME INTERVALS FOLLOWING A SINGLE INJECTION OF 0.06 MG. OF DIETHYLSTILBESTROL

Castrate Time (hours)

No. of animals

0 1

16 8 8

3 6 12 24 36 48 96

8 8 8 8 8 8

8

Average adrenal weight (mgs.)

Average body weight (Gms.)

8.1 8.9 7.7 7.5 6.9 7.3 7.3 6.9 6.7 6.5

135 122 115 112 99 107 118 105 103 111

Average mgs. ascorbic acid per 100 gms. adrenal wt. with S.E.M. 204±23.4 152±13.6 144 + 12.1 152±14.2 219± 3.9 199 + 11.6 199±23.4 191 ±19.4 207 ±10.3 201 ±11.1

V

value Normal .0574 .0232 .0588 .5287 Normal Normal Normal Normal Normal

Normal 0 * 1

3 6 12 48 96

16 12 20 12 8 8 8 4

12.9 12.8 12.5 13.1 12.2 13.6 ' 12.3 10.4

116 118

120 115 112

107 117 109

109± 95± 92± 117+ 111± 97± 98+ 112+

2.1 2.4 1.1 3.9 3.7 1.3 2.0 • 9.0

Normal .0001 .0001 .0719 Normal .0012 .0002 Normal

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May, 1952

VITAMIN C CHANGES AFTER GONADECTOMY

533

an abrupt fall within 30 minutes following the injection of stilbestrol. This was also the point of maximum depletion of ascorbic acid, a depletion of 63.2% of the zero hour percentage (Figure 2). The ascorbic acid content then increased so that by 6 hours it had returned to the level normal for the spayed animal and remained at this level for the duration of the experiment. There was no compensatory increase above normal as has been found in the intact hamster (Figures 1 and 2), the guinea pig and the rat (Long, 1947). The curve for the ascorbic acid values for spayed hamsters