Toxicology, 5 (1975) 43--47 © Elsevier/North-Holland, Amsterdam -- Printed in The Netherlands


N. GILLIAVOD and A. LI~ONARD Laboratory of Genetics, Department of Radiobiology C.E.N. --S.C.K., B-2400 Mol (Belgium)

(Received December 24th, 1974) (Accepted March 20th, 1975)

SUMMARY The testicle is k n o w n to be the critical organ in acute exposure of experimental mammals to cadmium. Such t r e a t m e n t results in t e m p o r a r y sterility and, very often, in the appearance of testicular interstitial cell tumors. The present experiments were p e r f o r m e d to determine w het her such deleterious effects on the male reproductive organs involved any genetic hazards for the surviving germ cells. The administration of 1.75 mg/kg cadmium chloride to male mice did n o t increase the d o m i n a n t lethals during the first three weeks after t r e a t m e n t and failed to induce translocation in the F 1 male off-spring. No c h r o m o s o m e rearrangement was observed in the treated males after i.p. injection of 0.5, 1.75 or 3.0 mg/kg cadmium chloride.

INTRODUCTION Although cadmium is n o t t h o u g h t to represent a hazard to man as importan t as lead or m e r c u r y it must nevertheless be considered a dangerous pollutant. The main sources of cadmium pollution are mines and metallurgy, chemical industries, scrap metal t r e a t m e n t , electroplating, superphosphate fertilizers and cadmium pesticides. Several cases of chronic cadmium poisoning have been r e p o r t e d in the literature [9--11,15,27] the Itai-itai disease which was endemic between 1940 and 1965 in the Zinzu River [10] being an e x t r e m e manifestation of such chronic cadmium poisoning. The most c o m m o n effect of cadmium exposure is renal tubular dysfunction, in m any cases indicated only by an increased excretion of proteins, while in more advanced cases there will also be disturbances in the metabolism of bone minerals [ 1 4 , 1 6 - - 1 8 , 2 2 , 2 3 ] . On the other hand [ 1 , 6 , 8 , 1 3 , 1 9 , 2 0 ] , the testicle has been d e m o n s t r a t e d to be the critical organ after acute exposure of experimental mammals, such t r e a t m e n t resulting in t e m p o r a r y sterility due to the destruction of spermatogonial cells. The present experiments were p e r f o r m e d to determine w het her such


d e l e t e r i o u s e f f e c t s on t h e m a l e r e p r o d u c t i v e organs involve a n y genetic h a z a r d s f o r t h e surviving g e r m cells. MATERIALS AND METHODS Male m i c e f r o m B A L B / c strain, aged 1 1 - - 1 3 w e e k s , w e r e injected i.p. w i t h 0.5, 1.75 or 3.0 m g / k g o f c a d m i u m chloride. O n e d a y a f t e r t r e a t m e n t each r e c i p i e n t o f 1.75 m g / k g was caged w i t h t h r e e virgin f e m a l e s f r o m t h e s a m e strain. A f t e r 7 and 14 d a y s t h e f e m a l e s w e r e r e p l a c e d b y fresh ones t o c o m p a r e t h e sensitivity o f s p e r m a t o z o a , m a t u r e a n d early s p e r m a t i d s . 10 d a y s later the f e m a l e s w e r e dissected a n d the n u m b e r o f c o r p o r a lutea, live a n d d e a d e m b r y o s c o u n t e d a n d c o m p a r e d w i t h t h o s e f r o m f e m a l e s m a t e d in t h e s a m e m a n n e r w i t h m a l e s w h i c h h a d received saline only. T h e t r e a t e d males w e r e killed t h r e e m o n t h s later a n d their testes e x a m i n e d b y m e a n s o f air-drying t e c h n i q u e o f Evans et al. [4] f o r t h e p r e s e n c e o f t r a n s l o c a t i o n c o n f i g u r a t i o n s [ 1 1 ] . F u r t h e r m o r e , t h e F 1 t r a n s l o c a t i o n test was p e r f o r m e d on 120 F 1 m a l e s sired d u r i n g t h e first t h r e e w e e k s a f t e r t r e a t m e n t b y B A L B / c m a l e m i c e given 1.75 m g / k g o f c a d m i u m chloride and m a t e d w i t h c o n t r o l females. In t h e s p e r m a t o c y t e test 100 cells w e r e a n a l y s e d p e r t r e a t e d m a l e ; in the F 1 t r a n s l o c a t i o n test, 25 p e r F] m a l e . RESULTS AND DISCUSSION T h e results o f t h e d o m i n a n t l e t h a l i t y test are s u m m a r i z e d in T a b l e I. T h e o n l y e f f e c t o b s e r v e d in the f e m a l e s m a t e d w i t h the t r e a t e d m a l e s was a slight b u t n o n - s i g n i f i c a n t decrease (t -- 0.6; 0.6 > P > 0.5) in t h e n u m b e r o f living e m b r y o s as well as a decrease in t h e rate o f p r e g n a n c y d u r i n g t h e t h i r d w e e k a f t e r t r e a t m e n t . E x a m i n a t i o n o f dividing s p e r m a t o c y t e s in t h e t r e a t e d ani-


Week Total of the females mating

% of Corpora preglutea per nant female females

Implantation per female

Live embryos per female

Dead embryos per female


1st 2nd 3rd

67 67 77

65 68 57

8.95 9.15 9.22

8.11 8.52 8.09

6.13 6.13 6.09

1.97 2.39 2.00


1st 2nd 3rd

67 67 86

68 70 45

8.89 9.19 8.79

8.06 7.82 7.46

6.15 5.87 5.41

1.91 1.95 2.05



Treatment (mg/kg)

Spermatocyte test

0 0.5 1.75 3.OO 1.75

F1 translocation test

Animals examined

Cells analysed

Cells with translocation configuration

10 10 10 10

1000 1000 1000 1000

0 0 0 0




mals or in the F1 male off-spring failed to reveal any c h r o m o s o m e rearrangem e n t such as reciprocal translocation (Table II). Very few experiments have been p e r f o r m e d up to now to determine the genetic hazards of cadmium to mammals. Cadmium chloride added to leukocy te cultures and to fibroblast cultures for 24, 38 or 72 h does n o t induce c h r o m o s o m e aberrations according to Pat t on and Allisson [ 2 1 ] , but Shiraishi et al. [25] observed several c h r o m o s o m e aberrations in cultured human leukocytes after t r e a t m e n t with cadmium sulfide at a final concentration of 6.2 • 10 - 2 pg/ml of culture fluid. These included chromatid and isochromatid breaks, translocations and dicentrics. Positive observations in vivo have been r e p o r t e d by Shiraishi and Yoshida [26] and D e k n u d t and L6onard [2] who observed severe c h r o m o s o m e aberrations such as dicentric and ring c h r o m o s o m e s in peripheral blood l eukocyt es of workers engaged in the m a n u f a c t u r e of cadmium. The fact t hat cadmium produces no genetic effects in male mouse germ cells is th er e f or e rather surprising since acute exposure to this metal has been shown to kill spermatogonia [ 1,6,8,13,19 ] and to induce testicular interstitial cell tumors in mice and rats [ 7 , 2 4 ] . Similar observations have been r ep o r ted with ot her chemicals such as m i t o m y c i n C which is c y t o t o x i c to spermatogonia (resulting in t e m p o r a r y sterility of treated males [ 1 2 ] ) but does n o t p roduce reciprocal translocations in premeiotic male germ cells [5] while having only a slight d o m i n a n t lethal effect in spermatocytes [ 3 ] . This observation demonstrates once more that the action of chemicals on mammalian reproductive organs differs markedly from t hat of ionizing radiation. ACKNOWLEDGEMENTS This work has been p e r f o r m e d with grant from the " F o n d s National de la Recherche Scientifique Medieale".


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Mutagenicity tests with cadmium in the mouse.

The testicle is known to be the critical organ in acute exposure of experimental mammals to cadmium. Such treatment results in temporary sterility and...
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