Genetic factors and acute carbon monoxide intoxication

The Science of the Total Enuironment, 23 (1982) 189--196 Elsevier Scientific Publishing Company, Amsterdam -- Printed in The Netherlands

GENETIC FACTORS AND ACUTE CARBONMONOXIDE INTOXICATION

Maurice STUPFEL~, Arthur PERRAMON~ , Victor-Hugo DEMARIA PESCE~, Philippe M~RAT~ , V6ronique GOURLETm, Huguette THIERRY~

mlnstitut National de la Sant6 et de la Recherche M~dicale (INSERM), Groupe de Recherches sur les M6canismes Physiopathologiques des Nuisances de l'Environnement, 44, Chemin de Ronde, 78110, Le Wsinet, France.

~ I n s t i t u t National de la Recherche Agronomique (INRA), Laboratoire de G~n~tique Factorielle, 78350, Jouy en Josas, France.

ABSTRACT. LD12:12 (L = 0600-1800; L = 100 lux) synchronized, 5-10 grouped small laboratory vertebrates, were during part of the L period (0900-1130) exposed to an acute carbon monoxide intoxication, resulting in an overall 50 %mortality. Interspecific and interstrain differences were found in outbred and inbred mice (CBA, C57Bl, OFI), outbred rats (Sprague-Dawley, Wistar), guinea-pigs, chicks (R+, R-), and Japanese quails. In this last bird species, a genetic selection produced two genotypes, one sensitive and one resistant to a carbon monoxide acute intoxication.

INTRODUCTION. Carbon monoxide is s t i l l a health hazard for homes, f i r e fighters, industry workers which causes thousands of death per year (10, 23), and i t is also a potential risk for patients witi~ cardiovascular disease (2, 25). I t is mostly believed, since Claude BERNARD, that carbon monoxide intoxication results from the CO fixation on hemoglobin (5), though the importance of its effects

0048-9697/82/0000--0000/$02.75 © 1982 Elsevier Scientific Publishing Company

189

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on r e s p i r a t o r y enzymes (6) is debated. Moreover tile role of genetic factors has been evidenced in other r e s p i r a t o r y cilallenges and p a r t i c u l a r l y in acute hypoxia ( I I ,

13, 19), which is u s u a l l y con-

sidered as the consequence of an exposure to high concentrations of carbon monoxide. I t is t h i s aspect of genetic v a r i a t i o n s in carbon monoxide t o x i c i t y that we have experimentally investigated (20, 21). Considering tile t o x i c i t y of carbon monoxide, the acute effects of high concentrations of t h i s gas have to be studied on animal models which advantages and l i m i t s have been discussed elsewhere (18). Two kinds of experimental processes were used to i n v e s t i g a t e possible genetic differences. The f i r s t lity

one was to t e s t the l e t h a -

of an acute CO t o x i c i t y in d i f f e r e n t species and several s t r a i n s

of small laboratory vertebrates. The second one was to t r y to select, in a same species, two s t r a i n s , one s e n s i t i v e and one r e s i s t a n t to an acute carbon monoxide i n t o x i c a t i o n . For t h i s l a s t experimentation Japanese q u a i l s were chosen because i t is possible, with t h i s bird species, to get a new generation w i t h i n six weeks. ANIMALS AND METHODS. Conventional albino guinea-pigs, outbred OFI , inbred CBA and C57BI, SPF ( s p e c i f i c pathogen free) mice, outbred Sprague-Dawley and Wistar, SPF, rats were issued from the Breeding Research Center of Les Oncins (Saint Germain sur l ' A r b r e s l e , 69210, France). R+ and R- young chickens which had been selected f o r a high and low adult feed e f f i c i e n c y , and Japanese q u a i l s which had been selected f o r resistance to carbon monoxide acute t o x i c i t y came from the Factor i a l Genetics Laboratory of INRA. The animals were, according to species, housed in separated rooms, which were a i r conditioned (temperature 20-21°C; humidity 50-60 %) and LDI2:I 2 (L = 0600-1800 hr) l i g h t e d with i00 l u x . A l l animals were randomly d i s t r i b u t e d , species, s t r a i n and sex being separated, in wire mesh cages of 2 sizes. In the smallest ones, mice were 10-11 grouped; in the largest ones, r a t s , guinea-pigs, q u a i l s , chicks were 5-7 grouped. A synthetic food appropriate to the various species, and tap water, were provided ad l i b i t u m . For the acute i n t o x i c a t i o n , the wire mesh cages containing the animals were enclosed in a l u c i t e parallelepiped chamber. A mixture

191

of 5.0 % carbon monoxide, 21% oxygen, and 74 % nitrogen (Compagnie FranGaise des Produits Oxyg~n~s) was flushed through t h i s chamber at a rate of 75-120 l i t e r s

per hour. Soda lime (Prolabo, anesthesia

grade) was placed in the bottom of the chamber to maintain a low concentration of carbon dioxide (0.05-0.30 %). The atmosphere of the chamber was analyzed f o r carbon monoxide, carbon dioxide (ONERA i n f r a r e d analysers) and f o r oxygen (Vial monitor) concent r a t i o n s . Carbon monoxide f l u s h i n g was stopped and the chamber was opened when i t appeared that about 50 % of the enclosed animals had ceased breathing. Continuous recording (Maxant recorder) shows that inside the chamber, during the i n t o x i c a t i o n , the temperature and humidity ranges uere r e s p e c t i v e l y 19-22°C and 84-100 %; the l i g h t i n g was 100-120 l u x . Because of circadian v a r i a t i o n s in s e n s i t i v i t y to carbon monoxide i n t o x i c a t i o n (15), a l l experiments were begun at the same time of the day (0910-0920). Statistical

differences were tested by Fisher t t e s t , variances I

a n a l y s i s , o r chi-square t e s t i n g . RESULTS. Table 1 shows the r e s u l t s of d i f f e r e n t experiments performed in one s t r a i n of guinea-pigs, 3 s t r a i n s of mice, 2 strains of r a t s , 2 s t r a i n s of chicks and 2 (selected f o r CO s u r v i v a l ) s t r a i n s of q u a i l s . Species, s t r a i n , sex, age, number of animals and t h e i r body weights, are shown in columns 1-6 of the table. In column 7 is the product of the length of tile carbon monoxide exposure ( i n minutes) by the pCO ( i n t o r r ) ,

which was achieved when the bv~rall 50 % survival

was reached. In column 8 are the survival percentages of the animals of each species, s t r a i n , and sex. When several r e s u l t s were obtained, the a r i t h m e t i c mean is followed by i t s standard deviation (mean ± SD). Interspecies comparisons, made by a c l a s s i f i c a t i o n according to the length of i n t o x i c a t i o n m u l t i p l i e d by pCO, give a c l a s s i f i cation ranging from small to great CO s e n s i t i v i t y : guinea-pigs < mice < rats = chicks < q u a i l s . When considering s t r a i n and sex differences the f o l l o w i n g data are obtained :

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Table

SURVIVAL TO AN OVERALL 50 % LETHAL CARBON MONOXIDE CHALLENGE OF DIFFERENT

1.

SPECIES AND STRAINS OF SMALL LABORATORY VERTEBRATES. SD; n u m b e r o f

(mean

SPECIES

:

STRAINS

GUINEAPIGS

: :

Conventional

:

:

~

:

:

:

: SEX : :: : :

: :

M F

: :

:

: :

CBA

: :

M F

experiments

:

: 47±7 :

: :

:

:

:

:

: :

: :

57 59

MICE

:

:

C57B1

:

: :

:

: :

: :

SpragueDawley

M F

M F

: 45+4

: :

:

:

:

: 368±60 : 354 ± 6 9

: 1494 + 1 0 3 ( 2 ) : -

: :

:

:

:

:

:

: :

: :

:

:

: :

19±1 15±2

: :

103 101

: :

M F

: :

19±2 16±2

: :

Wistar

:R+ :R-

: : : : : :

: :

M F

: :

M F

: Ls-

47.37 55.93

10.75 24.36

: : :

: :

65.05 67.35

:

:

:

:

:

:

: :

: :

: :

: :

:

:

:

: :

9.84 68.33

: :

35.00 91.67

: :

40.93 65.14

: :

58.23 71.26

103 102

: :

: :

27±2 21 ± 2

: :

61 62

: 351+-14 : 222+-10

: :

: :

60 60

: 349±12 : 207 ± 8

: :

: :

28±0

: :

105 109

: 193±34 : 203±30

: :

: :

: :

158 174

: 140±14 : 151±20

: :

: : :

58.25 84,31

158 +- 2 2 ( 4 )

: 56+_9 M F

285 + 6 ( 3 ) -

:

: M : : M :

Ls+

SURVIVAL %

:

79+-3 : :

QUAILS

: ":

:

RATS

CHICKS

: x END PCO : (rain. x torr)

WEIGHTS (g)

:

:

: :

:

:

: :

OFI

:

: LENGTH OF EXP.

:

93 78

: :

brackets) -:

A N I M A L S

AGE : NUMBERS : days: :

:

between

: : :

124

: 134±23

127

: 142±20

: :

166 ~ 3 0 ( 2 )

95 ± 3 1 ( 9 )

: : :

25.00 33.07

193

In mice, CBA of both sexes are s t a t i s t i c a l l y

( P < 0.01) less

r e s i s t a n t than C57BI and OFI . A sex-related difference is noted in OFI where survival is s t a t i s t i c a l l y (P < 0.001) less in males (58.25 %) than in females (84.31%). In rats,a sex-related difference (P < 0.001) is noticed in both strains. In each sex,Sprague-Dawleys are s i g n i f i c a n t l y (0.01< P <0.001) less r e s i s t a n t than Wistars. R+ chicks are s i g n i f i c a n t l y (P < 0.001) less r e s i s t a n t than R- chicks. In Japanese quails a s i g n i f i c a n t ( P < 0.001) difference is observed between the selected r e s i s t a n t Ls+ and the selected sensit i v e Ls- s t r a i n . In each of t h i s s t r a i n , males are s l i g h t l y (P < 0.05) less r e s i s t a n t than females.

DISCUSSION. The aforementioned data confirms the discrepancies in carbon monoxide acute t o x i c i t y according to the chosen animal model ( I , 4, 14, 24). In addition i t proves t h a t , in a same species, differences are obtained according to s t r a i n and sex. Several physiological and biochemical factors are involved into these reported genetic differences. A few concern v e n t i l a t i o n and d i f f u s i o n which determine the intake of the inhaled CO, o~he~sconcern cardiac output which takes care of i t s d i s t r i b u t i o n . Both these r e s p i r a t o r y and c i r c u l a t o r y parameters are grossly related to body weights (8, 16). To explain these i n t e r and intraspecies discrepancies in carbon monoxide t o x i c i t y one must also consider differences in COHb kinetics (7, 9, 21), in neurovegetative reactions (12), and even in r e s p i r a t o r y circadian and u l t r a d i a n rhythms (17, 22) of the various phenotypes. I t can be conceived that the genetic selection performed in quails has developed one or a few of the physiological or biochemical mechanisms considered above and responsible f o r resistance or sensitivity

to the acute carbon monoxide challenge.

Moreover, i f carbon monoxide i n t o x i c a t i o n might be considered as a kind of hypoxia, i t can be hypothesized that the here reported CO genetic survival differences are s i m i l a r to those which have been reported in experimental acute hypoxic hypoxia (3, i i ,

12, 13,

23), and which are well known in high and low a l t i t u d e residents of Andes and Himalaya.

194

Therefore the here evidenced genetic discrepancies, in several vertebrates species, in survival to an acute CO challenge, point out the p o s s i b i l i t i e s of such genetic differences in the human species. These genetic differences in s e n s i t i v i t y to carbon monoxide could be of interest to explain human interindividual discrepancies of cardiac pathological s e n s i t i v i t y of smokers and m o r t a l i t y in a i r pollution acute episodes.

ACKNOWLEDGEMENTS. We acknowledge with gratitude the s k i l l f u l technical assistance of Christian LEMERCERRE, Jean-Louis MONVOISIN, G~rard COQUERELLE, and the talented secretarial assistance of M i r e i l l e PAUCHARD.

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~

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