15.9. 1976
Specialia
t w e e n t o l u e n e / N N a O H s h o w e d t h a t over 99% of radioa c t i v i t y was p r e s e n t in t h e n e u t r a l fractions a n d o n l y 0.63-0.78% was associated w i t h phenolic fractions. The presence of e s t r o n e a n d estradiol-17/3 was e x c l u d e d in phenolic f r a c t i o n s f r o m b o t h precursors. Initial p a p e r c h r o m a t o g r a p h y of t h e n e u t r a l m e t a b o l i t e s f r o m progesterone i n c u b a t i o n s (in c y c l o h e x a n e : b e n z e n e (3: 1)p r o p y l e n e glycol s y s t e m ) r e s u l t e d in t h e s e p a r a t i o n of t h e m a t e r i a l into 4 zones of r a d i o a c t i v i t y . Zone 1 i n c l u d e d : p o l a r u n i d e n t i f i e d m a t e r i a l ; zone 2: 3e,20c~-dihydroxy5/%pregnane a n d 3/~,20~-dihydr0xy-5~-pregnane; zone 3: 3~-hydroxy-5/%pregnan-20-one a n d 3~-hydroxy-5flp r e g n a n - 2 0 - o n e ; zone 4: u n c o n v e r t e d p r o g e s t e r o n e a n d 5/%pregnan-3,20-dione. The c r y s t a l l i z a t i o n d a t a for t h e m e t a b o l i t e s i d e n t i f i e d are p r e s e n t e d in T a b l e I. The f o r m a t i o n of t e s t o s t e r o n e , 4 - a n d r o s t e n - 3 , 1 7 - d i o n e a n d 5fi-androstane d e r i v a t i v e s was e x c l u d e d b y c r y s t a l l i z a t i o n s t u d y . As can be seen f r o m Table II, p r o g e s t e r o n e was e x t e n s i v e l y r e d u c e d to 5/%pregnane d e r i v a t i v e s b y t h e p l a c e n t a e f r o m b o t h 15 a n d 28 d a y p r e g n a n t r a b b i t s . I t is also e v i d e n t t h a t t h e yield of p r e g n e n o l o n e c o n v e r s i o n was m u c h lower t h a n t h a t of p r o g e s t e r o n e . I n t h e n e u t r a l f r a c t i o n f r o m p r e g n e n o l o n e incubations, p r o g e s t e r o n e was isolated. Besides, 3/%hydroxy-5/3-pregnan-20-one a n d 5fi-pregnan-3,20-dione were also identified (Table I). Discussion. The p r e s e n t results s h o w t h a t r e d u c t i o n t o 5fl-pregnane d e r i v a t i v e s is t h e only m e t a b o l i c p a t h w a y of p r o g e s t e r o n e and p r e g n e n o l o n e in t h e p l a c e n t a of m i d g e s t a t i o n a n d n e a r t e r m r a b b i t s . E n z y m e activities dem o n s t r a t e d b y t h e isolation a n d i d e n t i f i c a t i o n of t h e
1213
p r e g n e n o l o n e and p r o g e s t e r o n e m e t a b o l i t e s are: A 53/%hydroxysteroid d e h y d r o g e n a s e , AS,A4-isomerase, 5flreductase, 3e-, 3/%, a n d 2 0 e - h y d r o x y s t e r o i d d e h y d r o genases. P r e g n e n o l o n e c o n v e r s i o n to p r o g e s t e r o n e , s h o w n in t h e p r e s e n t e x p e r i m e n t , c o n f i r m s t h e p r e v i o u s r e p o r t of MATSUMOTO et al. s. T h e lack of e s t r o g e n s y n t h e s i s in r a b b i t p l a c e n t a has a l r e a d y b e e n r e p o r t e d 9. Stereospecific 5/%reduction in r a b b i t p l a c e n t a is unique a m o n g s h o r t g e s t a t i o n species. P r e s e n t results are in c o n t r a s t to t h o s e r e p o r t e d in similar s t u d i e s w i t h m o u s e a n d r a t p l a c e n t a l q u a r t e r s a n d h o m o g e n a t e s , in w h i c h t h e t h e p r i n c i p a l m e t a b o l i t e s were s h o w n to be s a t u r a t e d 5~-pregnane d.er i v a t i v e s ~ 4. The m o s t p r o n o u n c e d difference in t h e distrib u t i o n of e n z y m a t i c activities in t h e p l a c e n t a e of v a r i o u s species a p p e a r s to lie in t h e absence or p r e s e n c e of 17h y d r o x y l a s e and 17-20 lyase activity. The c a p a c i t y for C19 s t e r o i d s y n t h e s i s f r o m C2~ precursors could n o t be found in r a b b i t p l a c e n t a , as was s h o w n earlier in m o u s e and r a t 2,a,s. These o b s e r v a t i o n s e m p h a s i z e once again t h e i m p o r t a n t differences w h i c h exist in t h e n a t u r e of t h e metabolites formed from the same precursors by the same organ f r o m different m a m m a l i a n species. 6 1. E. Bus~, The Chromatography o/ Steroids (Pergamon Press, Oxford 1961), p. 358. 7 L. R. AXELROD,C. MATTHIJSSEN, J. W. GOLDZlEHI~Rand J. E. PULLIAM, Acts endoer., Cnpenb. (1965), Suppl. 99. 8 1~. MATSUMOTO,G. YAMANE,H. ENDO, K. KOTOHand K. OKANO, Acta endocr., Copenh. 67, 577 (1969). " L. AINSWORTHand K. RYAN, Endocrinology 79, 875 (1966).
P r e - and P o s t - P a r t u m P l a s m a A m i n e O x i d a s e D i f f e r e n c e s in the R h e s u s M o n k e y (Macaca mulatta) 1 D. E. REDMOND, JRfl, J. BAULU a n d D. L. MURPHY
Department o/ Psychiatry, Yale University School o / M e d i c i n e , 3d P a r k Street, N e w H a v e n (Connecticut 06510, U S A ) and the Laboratory o/ Clinical Science, N . I . M . H . , and the Caribbean P r i m a t e Research Center at L a Parguera (Puerto Rico), 16 M a r c h 7976. S u m m a r y . P l a s m a a m i n e o:~idase a c t i v i t y increased f r o m 23.4 n m o l / m l / h d u r i n g p r e g n a n c y to 49.5 n m o l / m l / h d u r i n g a n e x t e n d e d p o s t p a r t u m period in 10 rhesus m o n k e y s . C o m p a r i s o n w i t h n o n - p r e g n a n t c o n t r o l m o n k e y s s a m p l e d a t similar t i m e s i n d i c a t e d t h a t t h e significant differences were in t h e e x t e n d e d p o s t p a r t u m period. M o n o a m i n e oxidase, a d e a m i n a t i n g e n z y m e (MAO, m o n o a m i n e : O~ o x i d o r e d u c t a s e d e a m i n a t i n g EC.1.4.3.4) was first discovered b y HARE ~ in 1928. Its m a j o r roles in deactivating and degrading monoamille neurotransmitters a n d a m i n e m e t a b o l i t e s , a n d in r e g u l a t i n g a m i n e s y n t h e s i s via a f e e d b a c k m e c h a n i s m , h a v e b e e n e x t e n s i v e l y reviewed4 7. Various i s o e n z y m e s are f o u n d associated w i t h m i t o c h o n d r i a in b o d y tissues, platelets, a n d n e r v e - e n d i n g p r e p a r a t i o n s f r o m b r a i n *-1~ A soluble e n z y m e f o u n d in t h e p l a s m a has d i f f e r e n t c o - f a c t o r r e q u i r e m e n t s a n d s u b s t r a t e - i n h i b i t o r responses f r o m t h e m i t o c h o n d r i a l M A O ' s f o u n d in all o t h e r tissues. T h e p l a s m a e n z y m e h a s t h e h i g h e s t a f f i n i t y for b e n z y l a m i n e n , ~2, a l t h o u g h t y r a m i n e has b e e n t h e m o s t f r e q u e n t l y s t u d i e d s u b s t r a t e . P r e v i o u s i n v e s t i g a t o r s h a v e o b s e r v e d lower p l a s m a MAO activities in w o m e n t a k i n g s y n t h e t i c p r o g e s t i n - c o n t a i n i n g p h a r m a c o l o g i c a g e n t s t h a n in a g e - m a t c h e d c o n t r o l s ~a. One s t u d y f o u n d decreases in p l a s m a MAO f r o m an
t We thank Dr. C. H. CONAWAYand Dr. J. D. LoY of the Caribbean Primate Research Center and J. G. VA~DENB*RGHand W. POST of the North Carolina Department of Mental Health for the use of their facilities and animals. We also acknowledge the technical
assistance of C. H. DONNELLY and Ms. SUZANNE BAULU. This work was supported by a grant from Dr. H. C. ROBINSON,JR., through Yale University Medical School and by NIH Contract No. 71-2003. Address reprint requests to: D. E. REDMOND,JR., Department of Psychiatry, 34 Park Street, New Haven, Connecticut 06510, USA. a M. L. C. HARE, Biochem. J. 22, 986 (1928). I. J. KOPIN, Pharmac. Rev. 76, 179 (1964). 5 F. COSTA and P. GREENGARD,Advances in Biochemical Psychopharmacology (Raven Press, New York 1972}, vol. 5. 6 N. WEINER and R. BJUR, ill Advances in Biochemical Psycho ~ pharmacology, (Eds. F. COSTA and P. GREENGARD; Raven Press, New York 1972), voI. 5, p. 409. v D. L. MURPHY, C. H. DONNELLY, and M. ]3UCHSBAUiVI,in preparation (1976). s j. p. JOHNSTON, Bioehem. Pharmac. 77, 1285 (1968). 9 N. H. Nz~" and C. GORIDIS, in Advances in Biochemical Psychopharmacology (Eds. F. COSTA and P. GREENGARO;Raven Press, New York 1972), voh 5, p. 307. C. H. DONNELLu and D. L. MURPHY,in preparation (1976). C. M. MCEWEN, JR., J. biol. Chem. 240, 2003 (1965). C. M. MCEWEN, JR., in Advances in Biochemical Psychopharmaeology (Eds. F. COSTA and P. GREENGARD; Raven Press, New York 1972), vol. 5, p. 151. N. TRYDING, S. E. NILSSON, G. TIFVESSON, B. BERG, S. CARLSTRO.~, B. ELMFORS, and J. E. NILSSON, Scand. J. din. Lab. Invest. 23, 79 (1969).
1214
Specialia
a v e r a g e of 6 h before d e l i v e r y to a n a v e r a g e of 78 h a f t e r 1~. H o w e v e r , t h e r e was no difference b e t w e e n p l a s m a MAO a c t i v i t y in 3rd t r i m e s t e r p r e g n a n t h u m a n females a n d c o n t r o l s in a n o t h e r s t u d y 15. R e c e n t studies h a v e s u g g e s t e d tile p o s s i b i l i t y t h a t t h e t y r a m i n e - d e a m i n a t i n g e n z y m e a c t i v i t i e s m e a s u r e d in t h e l a t t e r s t u d y a f t e r c e n t r i f u g a t i o n a t 1000-2000 g reflected r e s i d u a l p a r t i c u l a t e M A O act i v i t y f r o m plateletsl6,17. C o n s i s t e n t w i t h t h i s interpretation, the plasma MAO changes previously reported in a s s o c i a t i o n w i t h t h e m e n s t r u a l cycle in w o m e n 15,1s were n o t f o u n d b y BELMAKER et al. ~9 in p l a s m a e n z y m e t h a t h a d b e e n c e n t r i f u g e d a t 80,000 g to r e m o v e r e s i d u a l p a r t i c u l a t e e n z y m e a c t i v i t y . C h a n g e s were n o t e d i n s t e a d i n t h e p l a t e l e t MAO. W e h a v e r e p l i c a t e d t h e s e f i n d i n g s in t h e female r h e s u s m o n k e y d u r i n g t h e m e n s t r u a l cycle 3% T h e p r e s e n t s t u d y was u n d e r t a k e n t o i n v e s t i g a t e t h e possible effects of p r e g n a n c y o n p l a s m a M A O a c t i v i t y in t h e r h e s u s m o n k e y , u s i n g h i g h speed c e n t r i f u g a t i o n a n d specific i n h i b i t o r s to r e m o v e possible c o n t a m i n a t i n g m i t o chondrial enzyme. Materials and methods. 10 h e a l t h y p r e g n a n t a d u l t female r h e s u s m o n k e y s (Macaca mulatta) were s t u d i e d f r o m t h e Caribbean Primate Research Center at La Parguera. Their m e a n c e n s u s - d e t e r m i n e d age was 9.1 years ( • 1.8 SEM). A n e q u a l n u m b e r of n o n - p r e g n a n t females were s t u d i e d as controls, aged 8.7 y e a r s ( :L 1.6 SEM). 16 of t h e s e a n i m a l s were enclosed in 1/2 or 1/4 acre corrals w h e r e t h e y were fed P u r i n a m o n k e y c h o w a n d fresh w a t e r a t a single feeder a d l i b i t u m . 4 c o n t r o l m o n k e y s were i n d i v i d u a l l y caged. M o n k e y s were c a u g h t in n e t s a n d h a n d r e s t r a i n e d w i t h o u t a n e s t h e t i c s while 10 c m ~ blood was d r a w n v i a a s a p h e n o u s v e n i p u n c t u r e i n t o s o d i u m h e p a r i n i z e d syringes. T h e s p e c i m e n s were d r a w n on e a c h a n i m a l d u r i n g t h e l a t t e r p a r t of p r e g n a n c y , a f t e r delivery, or a t c o r r e s p o n d ing t i m e s of t h e y e a r for controls, a n a v e r a g e of a p p r o x i m a t e l y 4 m o n t h s a p a r t . S p e c i m e n s were p l a c e d in acid c i t r a t e d e x t r o s e (ACD) p r e s e r v a t i v e a n d cooled in a n ice b a t h p r i o r to p r e p a r a t i o n . T h e s p e c i m e n s were c e n t r i f u g e d a t 2000 g for 3 0 + m i n a t 4~ a n d p l a s m a was r e m o v e d a n d frozen o n d r y ice w i t h i n 3 h of drawing. P l a s m a was s t o r e d a t - 2 0 ~ u n t i l analysis. P l a s m a M A O was determ i n e d u s i n g a m o d i f i c a t i o n of t h e m e t h o d d e s c r i b e d b y ROBINSON e t al. ~1 u s i n g 14C-benzylamine (10 -4 M) as s u b s t r a t e . V a l u e s are r e p o r t e d as n a n o m o l e s of d e a m i n a t ed p r o d u c t / m l p l a s m a / h . A t w o - f a c t o r a n a l y s i s of v a r i a n c e w i t h r e p e a t e d m e a s u r e s on one f a c t o r was used to a n a l y z e t h e d a t a , a f t e r t e s t s of t h e h o m o g e n i e t y of v a r i a n c e s in t h e p a r t i t i o n e d e r r o r t e r m s . T e s t s of simple m a i n effects completed the analysis after a significant AB interaction was f o u n d 23 Results. P l a s m a a m i n e oxidase a c t i v i t y i n c r e a s e d in e a c h of t h e p r e g n a n t a n i m a l s f r o m t h e s p e c i m e n d u r i n g t h e p r e g n a n c y to t h e p o s t d e l i v e r y s p e c i m e n ( F = 19.1, p < 0.001). T h e r e p e a t e d m e a s u r e was n o t s i g n i f i c a n t l y d i f f e r e n t in t h e c o n t r o l s ( F = 0.2, N.S.). T h e p r e p a r t u l n m e a n was n o t s i g n i f i c a n t l y d i f f e r e n t f r o m t h e c o r r e s p o n d ing t i m e s p e c i m e n for t h e c o n t r o l s ( F -- 2.3, N.S.), while t h e p o s t - p a r t u m m e a n was s i g n i f i c a n t l y d i f f e r e n t ( F 130, p < 0.001) (Table). Discussion. T h e differences in t h i s s t u d y are b e t w e e n p o s t p a r t u m MAO a c t i v i t i e s a n d c o r r e s p o n d i n g a c t i v i t i e s f r o m n o r m a l n o n - p r e g n a n t females w i t h o u t i n f a n t s in t h a t b i r t h season, a n d b e t w e e n p o s t p a r t u m v a l u e s a n d t h e v a l u e s f r o m p r e v i o u s s p e c i m e n s d u r i n g p r e g n a n c y in t h e s a m e animals. H o r m o n a l c h a n g e s in t h e p o s t - p a r t u m p e r i o d or associated w i t h n u r s i n g i n f a n t s m a y be i n v o l v e d in t h e s e differences. Some s o c i a l l y - i n d u c e d stress r e l a t e d to b e i n g t h e m o t h e r of a y o u n g i n f a n t in a p r i m a t e social g r o u p m i g h t also be i m p l i c a t e d , a l t h o u g h we k n o w of no b e h a v i o r a l data. to s u p p o r t t h i s possibility.
EXPERIENTIA 32/9
T h e r e is e v i d e n c e f r o m o t h e r s t u d i e s t h a t m i t o c h o n d r i a l MAO a c t i v i t y in a v a r i e t y of tissues is affected b y t h e e s t r u s cycle38 or b y p r e - t r e a t m e n t w i t h estradiol or progesterone ~*. W e h a v e n o t e d effects of t h e m e n s t r u a l cycle a n d of o v a r i e c t o m y on p l a t e l e t M A O in rhesus m o n k e y s , using t y r a m i n e as s u b s t r a t e , b u t f o u n d no c h a n g e s in p l a s m a a m i n e oxidase, u s i n g benzylamineS~ T h e r e s u l t s r e p o r t e d here a p p e a r to s u p p o r t t h e f i n d i n g of n o difference in p l a s m a MAO a c t i v i t y , m e a s u r e d w i t h t y r a m i n e as s u b s t r a t e , in p r e g n a n t h u m a n females a n d c o n t r o l s b y BRIGGS a n d BRIGGS 15. O u r r e s u l t s are n o t c o n s i s t e n t w i t h t h e findings of TRYDING et al. la w h o f o u n d r e d u c e d p l a s m a MAO a c t i v i t y in h u m a n females t a k i n g s y n t h e t i c prog e s t i n - c o n t a i n i n g p h a r m a c o l o g i c agents, w h e t h e r a m i n e oxidase a c t i v i t y was m e a s u r e d w i t h b e n z y l a m i n e or t y r a m i n e as s u b s t r a t e . T h e s m a l l differences in t h e op-
P l a s m a MAO a c t i v i t y (nmoles of b e n z y l a m i n e o x i d i z e d / m l p l a s m a / h ) in p a i r e d s p e c i m e n s f r o m 10 female r h e s u s m o n k e y s a n d c o r r e s p o n d ing specimens f r o m n o n - p r e g n a n t c o n t r o l s Monkey No.
Monkey No.
Mothers Pre-
Post-partum
095 231 R65 060 0Y6 0Z1 143 685 0X3 834
7.1 10.4 23.1 39.5 17.8 13.0 67.2 34.3 11.7 9.8
16.7 17.3 31.4 121.3 50.8 19.3 81.5 85.5 25.3 45.7
Mean S.D. S;E.M.
23.4 18.8 6.0
49.5 35.6 11.3
303 075 288 08D 09G 09C 0L9 283 309 241
Controls Spec. 1
Spec. 2
25.5 13.0 5.8 21.4 48.2 24.7 22.9 36.0 43.1 27.7
23.3 21.5 13.9 31.9 29.4 24.3 15.0 30.8 28.8 18.7
26.8 12.9 4.1
23.8 6.5 2.0
Mean No. days from Delivery =82.3 +30.7 S.E.M. 8.4 7.9
1, N. J. GILMORE, D. S. IROBINSON, A. NIES, D. SYLVESTER a n d C. J . RAVARIS, J. P s y e h o s o m . Res. 15, 215 (1971). 15 M. BRIGGS a n d M. BRIGGS, J . IReprod. F e r t . 29, 447 (1972). 16 E. A. ZELLER, B. H . BABU, M. J. CAVANAUGFI a n d G. H. STANICH, P h a r m a c . ires. C o m m u n . 7, 20 (1969). 17 D. L. MURPHY a n d C. H. DONNELLY, in Neuropsychopharmacology o/ Monoamines and their Regulatory Enzymes ( R a v e n Press, N e w Y o r k 1974), p. 71. 18 E. g . KLAIBER, Y. ]5~OBAYASHI,D. M. J~ROVERMAN a n d F. HALL, J. clin. E n d o c r i n o l o g y 33, 630 (1971). 19 IR. H . ]~BLMAKER, D. L. MURPHY, IR. J . WYATT a n d D. L. L o RIAUX, A r c h . gen. P s y c h . 37, 553 (1974). 20 D. E. REDMOND, JR., D. L. MURPHY, J. BAULU, M. G. ZEIGLER a n d C. R. LAKE, P s y c h o s o m . Med. 37, 417 (1975). 21 D. S. ROBINSON, W. LOVENBERG, H . KEISER a n d J . 8JOERDSMA, Bioehem. P h a r m a e . 77, 109 (1968).
33 B. J. WINER, Statistical Principles in Experimental Design, 2nd edn. McGraw Hill, New York 1962), p. 514. 33 A. J. ZOLOVICK, IR. PEARSE, K. W. BOEHLKE and B. E. ELEFTHERIOU, Science 754, 649 (1966). 34 G. C. S. COLLINS, M. SANDLER, E. D. WILLIAMS, a n d M. B. H . YOUDIM, N a t u r e , L o u d . 225, 817 (1970).
15.9. 1976
25 H. THOMS, in Classical Contributions to Obstetrics and Gynecology (Charles C. Thomas, Springfield 1935). 26 B. C. STEVENS, Soc. Set. Med. 5, 527 (1971). 27 C. PROTHI~ROE, Br. J. Psychiat. 115, 9 (1969). 2s G. C. ROSENWALD and M. W. STONEHILL, Psyehosom, Med. 3d, 129 (1972). 29 K. DALTON, Proc. R. Soc. Med. 6d, 1249 (1971). a0 W. A. BRown and P. SHERESItEFSKY, Psychiatry 3d, 139 (1972). 31 A. NIES, D. S. ROBINSON, C. L. RAVARIS and J. M. DAvis, Psychosom. Med. 33, 470 (1971). 32 D. L. MURPHY and R. WEiss, Am. J. Psychiatr. 128, 1351 (1972). 33 j . BOCKAR, R. H. ROTH and G. HENINGER, Life Sci. 15,2109 (1974). 34 D. L. MURPhy and R. J. WYATT, Nature, Lond. 238, 225 (1972). 35 R. J. WYATT, ~). L. MURPHY, R. BELMAKER, S. COHEN, C. H. DONNEZLY and W. POLLIN, Science 179, 916 (1973). 3s A. NIES, D. S. ROmNSON, L. S. HARRIS and K. R. LAMBORN,Adv. Biochem. Psyehopharmac. 72, 59 (1974). 37 H. M~LTZER and S. M. STAHL, Res. Commun. Chem. Path. Pharmac. 7, 419 (1974).
Thyroid
1215
Specialia
Hormone
in Serum
p o s i t e d i r e c t i o n f o u n d i n t h e s t u d y b y GILMORE e t al. 14 w e r e f r o m s a m p l e s d r a w n f r o m 6 h b e f o r e t o 134 h a f t e r delivery, a short time period for which we have no comparable data. T h e s i g n i f i c a n c e of p l a s m a a m i n e o x i d a s e a c t i v i t y a n d i t s r e l a t i o n s h i p s t o b r a i n or s y m p a t h e t i c m o n o a m i n e f u n c t i o n is n o t k n o w n . D i f f e r e n c e s i n e n z y m e s r e l a t e d t o t h e m o n o a m i n e s y s t e m s a r e of i n t e r e s t b e c a u s e of p o s sible relevance to the behavioral changes associated with t h e p o s t p a r t u m p e r i o d in h u m a n s s u c h as t h e p u e r p e r a l p s y c h o s e s w h i c h h a v e b e e n r e p o r t e d s i n c e H i p p o c r a t e s 25. T h e r e is a d i s a g r e e m e n t a s t o w h e t h e r t h e s e p s y c h o s e s represent distinct nosological entities or whether factors associated with the puerperal period precipitate the exp r e s s i o n of e p i s o d e s of s c h i z o p h r e n i c o r m a n i c - d e p r e s s i v e p s y c h o s i s 2 6 - 3 0 , b o t h of w h i c h h a v e b e e n a s s o c i a t e d w i t h c h a n g e s in o t h e r m o n o a m i n e e n z y m e activities31-37.
of Fetal Calf and Pregnant
Cow During
the Last Trimester
of Pregnancy
V. STRB~K 1, F. TOMgiK2 a n d I. H L O ~ N E K 3
Institute o/ Experimental Endocrinology, Slovak Academy o[ Sciences, Kramdre, Vldrska 3, CS-809 36 Bratislava (Czechoslovakia), University School o[ Veterinary Medicine, Brno (Czechoslovakia), and Institute o[ Molecular Genetics, Czechoslovak Academy o] Sciences, Praha (Czechoslovakia), 2 March 1976. Summary. C o n s i d e r a b l y h i g h e r t h y r o x i n e a n d t r i i o d o t h y r o n i n e c o n c e n t r a t i o n s in s e r a of b o v i n e f e t u s e s t h a n i n m a t e r n a l s a m p l e s w e r e f o u n d d u r i n g t h e l a s t t r i m e s t e r of p r e g n a n c y . I t w a s s h o w n t h a t m a m m a l i a n f e t a l t h y r o i d is a c t i v e in utero and that negative feedback control appears to be f u n c t i o n a l b e f o r e p a r t u r i t i o n 4,~. T o k n o w m o r e a b o u t mother-fetus interrelation, we compared thyroid hormone c o n t e n t in s e r a of b o v i n e f e t u s e s a n d p r e g n a n t c o w s d u r i n g t h e l a s t t h i r d of p r e g n a n c y . I Material and methods. 23 p r e g n a n t C z e c h r e d - w h i t e c o w s in t h e 2 4 1 - 2 7 9 t h d a y of g e s t a t i o n w e r e c l i n i c a l l y and hematologically examined before experiment. They were submitted to high epidural anaesthesia with procain, t h e h e a d of f e t u s w a s e x t e r i o r i z e d , a r t e r i a c a r o t i s w a s c a n n u l a t e d a n d t h e f e t u s w a s e x a n g u i n a t e d s. S a m p l e s of maternal blood were withdrawn during surgical procedure. In maternal and fetal sera, thyroxine was determined by radioimmunological method ~with some modifications o f F6LDESS a n d t r i i o d o t h y r o n i n e w a s d e t e r m i n e d a c c o r d -
i n g t o NAUMAN a n d •AUMAN 9. T h y r o x i n e a n t i b o d y w a s p r o d u c e d a n d k i n d l y s u p p l i e d b y D r . F6LDES s, t r i i o d o t h y r o n i n e a n t i b o d y w a s t h e k i n d g i f t f r o m D r . NAUMAN o. Results and discussion. T h e r e s u l t s a r e s u m m a r i z e d i n the Table. Both thyroid hormones, thyroxine and trii o d o t h y r o n i n e a r e in a t l e a s t t w i c e a s h i g h in c o n c e n t r a t i o n in f e t a l s e r a a s in m a t e r n a l o n e s . S t u d e n t ' s t a n d p a i r e d tests are highly significant. We did not find any correlation between thyroid hormone values in fetuses and mothers, between thyroid hormone content in sera and body w e i g h t of f e t u s e s . W e also d i d n o t f i n d a n x t e n d e n c y of thyroid hormone concentrations to change according to a g e of f e t u s e s . I n f e t u s e s , u n l i k e m o t h e r s , p o s i t i v e c o r relation between thyroxine and triiodothyronine was noted. In the sheep fetus, high thyroxine secretion rate in c o m p a r i s o n t o m a t e r n a l o n e w a s d e s c r i b e d 1~ HA-
Thyroxine (Tt) and triiodothyronine (T3) concentrations in maternal and fetal sera. BW, body weight; n.s., non-significant
T 4 (nmol/1) T 3 (nmol/l)
Mothers
Fetuses
n
n
Means 4- SE
23 42.9 4- 3.3 19 0.50 4- 0.05
Correlation coefficients T 4 (mother: fetus) T a (mother: fetus)
23 109.4 4- 5.1 19 1.22 :J= 0.15
+ 0.07767 + 0.21016
Fetal T4:BW Ta:BW T4:T 3 Maternal T4:T 3
-- 0.42385
P < 0.001 < 0.01 n.s. n.s.
23 19 19 19
Means4- SE
+0.20033 +0.00243 + 0.45442
ms. n.s. < 0.05 n.s.
1 Acknowledgment. We wish to thank Miss R. FAJKOgOVs for skilled technical assistance. 2 University School of Veterinary Medicine, Brno, Czechoslovakia. 3 Institute of molecular Genetics, Czechoslovak Academy of Sciences, Praha, Czechoslovakia. G. t3. HEGSTAD and L. J. WELLS, Anat. Rec. 718, 389 (1954). 5 M. NIKITOWITCtt and E. KNOBIL, J. clin. Endocr. Metab. 75, 837 (1955). s F. ToragiK and L. CERN'~, In vitro v r 2/3, 46 (1973). 7 S. J. SIEGEL, W. F. LINE, N. S. YANG, A. KWONG and C. ]FRANK, J. olin. Endocr. Metab. 37, 526 (1973). s O. F6LDES, to be published. Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava. 9 A. NAUMANand J. NAUMAN, Polskie Arehw. Med. wewn. 50, 1295 (1973). 10 p. W. NATHAmELSZ, R. S. COMLINE, M. SILVER and A. L. THOMAS, J. Endocr. 58, 535 (1973). n A. ERENBER~ and D. A. FISCHER, in Foetal and Neonatal Physiology (Eds. R. S. CO,LINE, K. W. CRoss, G. S. DAVES and P. W. NATHAmELSZ, Cambridge University Press, London 1973), p. 508.
Specialia
t w e e n t o l u e n e / N N a O H s h o w e d t h a t over 99% of radioa c t i v i t y was p r e s e n t in t h e n e u t r a l fractions a n d o n l y 0.63-0.78% was associated w i t h phenolic fractions. The presence of e s t r o n e a n d estradiol-17/3 was e x c l u d e d in phenolic f r a c t i o n s f r o m b o t h precursors. Initial p a p e r c h r o m a t o g r a p h y of t h e n e u t r a l m e t a b o l i t e s f r o m progesterone i n c u b a t i o n s (in c y c l o h e x a n e : b e n z e n e (3: 1)p r o p y l e n e glycol s y s t e m ) r e s u l t e d in t h e s e p a r a t i o n of t h e m a t e r i a l into 4 zones of r a d i o a c t i v i t y . Zone 1 i n c l u d e d : p o l a r u n i d e n t i f i e d m a t e r i a l ; zone 2: 3e,20c~-dihydroxy5/%pregnane a n d 3/~,20~-dihydr0xy-5~-pregnane; zone 3: 3~-hydroxy-5/%pregnan-20-one a n d 3~-hydroxy-5flp r e g n a n - 2 0 - o n e ; zone 4: u n c o n v e r t e d p r o g e s t e r o n e a n d 5/%pregnan-3,20-dione. The c r y s t a l l i z a t i o n d a t a for t h e m e t a b o l i t e s i d e n t i f i e d are p r e s e n t e d in T a b l e I. The f o r m a t i o n of t e s t o s t e r o n e , 4 - a n d r o s t e n - 3 , 1 7 - d i o n e a n d 5fi-androstane d e r i v a t i v e s was e x c l u d e d b y c r y s t a l l i z a t i o n s t u d y . As can be seen f r o m Table II, p r o g e s t e r o n e was e x t e n s i v e l y r e d u c e d to 5/%pregnane d e r i v a t i v e s b y t h e p l a c e n t a e f r o m b o t h 15 a n d 28 d a y p r e g n a n t r a b b i t s . I t is also e v i d e n t t h a t t h e yield of p r e g n e n o l o n e c o n v e r s i o n was m u c h lower t h a n t h a t of p r o g e s t e r o n e . I n t h e n e u t r a l f r a c t i o n f r o m p r e g n e n o l o n e incubations, p r o g e s t e r o n e was isolated. Besides, 3/%hydroxy-5/3-pregnan-20-one a n d 5fi-pregnan-3,20-dione were also identified (Table I). Discussion. The p r e s e n t results s h o w t h a t r e d u c t i o n t o 5fl-pregnane d e r i v a t i v e s is t h e only m e t a b o l i c p a t h w a y of p r o g e s t e r o n e and p r e g n e n o l o n e in t h e p l a c e n t a of m i d g e s t a t i o n a n d n e a r t e r m r a b b i t s . E n z y m e activities dem o n s t r a t e d b y t h e isolation a n d i d e n t i f i c a t i o n of t h e
1213
p r e g n e n o l o n e and p r o g e s t e r o n e m e t a b o l i t e s are: A 53/%hydroxysteroid d e h y d r o g e n a s e , AS,A4-isomerase, 5flreductase, 3e-, 3/%, a n d 2 0 e - h y d r o x y s t e r o i d d e h y d r o genases. P r e g n e n o l o n e c o n v e r s i o n to p r o g e s t e r o n e , s h o w n in t h e p r e s e n t e x p e r i m e n t , c o n f i r m s t h e p r e v i o u s r e p o r t of MATSUMOTO et al. s. T h e lack of e s t r o g e n s y n t h e s i s in r a b b i t p l a c e n t a has a l r e a d y b e e n r e p o r t e d 9. Stereospecific 5/%reduction in r a b b i t p l a c e n t a is unique a m o n g s h o r t g e s t a t i o n species. P r e s e n t results are in c o n t r a s t to t h o s e r e p o r t e d in similar s t u d i e s w i t h m o u s e a n d r a t p l a c e n t a l q u a r t e r s a n d h o m o g e n a t e s , in w h i c h t h e t h e p r i n c i p a l m e t a b o l i t e s were s h o w n to be s a t u r a t e d 5~-pregnane d.er i v a t i v e s ~ 4. The m o s t p r o n o u n c e d difference in t h e distrib u t i o n of e n z y m a t i c activities in t h e p l a c e n t a e of v a r i o u s species a p p e a r s to lie in t h e absence or p r e s e n c e of 17h y d r o x y l a s e and 17-20 lyase activity. The c a p a c i t y for C19 s t e r o i d s y n t h e s i s f r o m C2~ precursors could n o t be found in r a b b i t p l a c e n t a , as was s h o w n earlier in m o u s e and r a t 2,a,s. These o b s e r v a t i o n s e m p h a s i z e once again t h e i m p o r t a n t differences w h i c h exist in t h e n a t u r e of t h e metabolites formed from the same precursors by the same organ f r o m different m a m m a l i a n species. 6 1. E. Bus~, The Chromatography o/ Steroids (Pergamon Press, Oxford 1961), p. 358. 7 L. R. AXELROD,C. MATTHIJSSEN, J. W. GOLDZlEHI~Rand J. E. PULLIAM, Acts endoer., Cnpenb. (1965), Suppl. 99. 8 1~. MATSUMOTO,G. YAMANE,H. ENDO, K. KOTOHand K. OKANO, Acta endocr., Copenh. 67, 577 (1969). " L. AINSWORTHand K. RYAN, Endocrinology 79, 875 (1966).
P r e - and P o s t - P a r t u m P l a s m a A m i n e O x i d a s e D i f f e r e n c e s in the R h e s u s M o n k e y (Macaca mulatta) 1 D. E. REDMOND, JRfl, J. BAULU a n d D. L. MURPHY
Department o/ Psychiatry, Yale University School o / M e d i c i n e , 3d P a r k Street, N e w H a v e n (Connecticut 06510, U S A ) and the Laboratory o/ Clinical Science, N . I . M . H . , and the Caribbean P r i m a t e Research Center at L a Parguera (Puerto Rico), 16 M a r c h 7976. S u m m a r y . P l a s m a a m i n e o:~idase a c t i v i t y increased f r o m 23.4 n m o l / m l / h d u r i n g p r e g n a n c y to 49.5 n m o l / m l / h d u r i n g a n e x t e n d e d p o s t p a r t u m period in 10 rhesus m o n k e y s . C o m p a r i s o n w i t h n o n - p r e g n a n t c o n t r o l m o n k e y s s a m p l e d a t similar t i m e s i n d i c a t e d t h a t t h e significant differences were in t h e e x t e n d e d p o s t p a r t u m period. M o n o a m i n e oxidase, a d e a m i n a t i n g e n z y m e (MAO, m o n o a m i n e : O~ o x i d o r e d u c t a s e d e a m i n a t i n g EC.1.4.3.4) was first discovered b y HARE ~ in 1928. Its m a j o r roles in deactivating and degrading monoamille neurotransmitters a n d a m i n e m e t a b o l i t e s , a n d in r e g u l a t i n g a m i n e s y n t h e s i s via a f e e d b a c k m e c h a n i s m , h a v e b e e n e x t e n s i v e l y reviewed4 7. Various i s o e n z y m e s are f o u n d associated w i t h m i t o c h o n d r i a in b o d y tissues, platelets, a n d n e r v e - e n d i n g p r e p a r a t i o n s f r o m b r a i n *-1~ A soluble e n z y m e f o u n d in t h e p l a s m a has d i f f e r e n t c o - f a c t o r r e q u i r e m e n t s a n d s u b s t r a t e - i n h i b i t o r responses f r o m t h e m i t o c h o n d r i a l M A O ' s f o u n d in all o t h e r tissues. T h e p l a s m a e n z y m e h a s t h e h i g h e s t a f f i n i t y for b e n z y l a m i n e n , ~2, a l t h o u g h t y r a m i n e has b e e n t h e m o s t f r e q u e n t l y s t u d i e d s u b s t r a t e . P r e v i o u s i n v e s t i g a t o r s h a v e o b s e r v e d lower p l a s m a MAO activities in w o m e n t a k i n g s y n t h e t i c p r o g e s t i n - c o n t a i n i n g p h a r m a c o l o g i c a g e n t s t h a n in a g e - m a t c h e d c o n t r o l s ~a. One s t u d y f o u n d decreases in p l a s m a MAO f r o m an
t We thank Dr. C. H. CONAWAYand Dr. J. D. LoY of the Caribbean Primate Research Center and J. G. VA~DENB*RGHand W. POST of the North Carolina Department of Mental Health for the use of their facilities and animals. We also acknowledge the technical
assistance of C. H. DONNELLY and Ms. SUZANNE BAULU. This work was supported by a grant from Dr. H. C. ROBINSON,JR., through Yale University Medical School and by NIH Contract No. 71-2003. Address reprint requests to: D. E. REDMOND,JR., Department of Psychiatry, 34 Park Street, New Haven, Connecticut 06510, USA. a M. L. C. HARE, Biochem. J. 22, 986 (1928). I. J. KOPIN, Pharmac. Rev. 76, 179 (1964). 5 F. COSTA and P. GREENGARD,Advances in Biochemical Psychopharmacology (Raven Press, New York 1972}, vol. 5. 6 N. WEINER and R. BJUR, ill Advances in Biochemical Psycho ~ pharmacology, (Eds. F. COSTA and P. GREENGARD; Raven Press, New York 1972), voI. 5, p. 409. v D. L. MURPHY, C. H. DONNELLY, and M. ]3UCHSBAUiVI,in preparation (1976). s j. p. JOHNSTON, Bioehem. Pharmac. 77, 1285 (1968). 9 N. H. Nz~" and C. GORIDIS, in Advances in Biochemical Psychopharmacology (Eds. F. COSTA and P. GREENGARO;Raven Press, New York 1972), voh 5, p. 307. C. H. DONNELLu and D. L. MURPHY,in preparation (1976). C. M. MCEWEN, JR., J. biol. Chem. 240, 2003 (1965). C. M. MCEWEN, JR., in Advances in Biochemical Psychopharmaeology (Eds. F. COSTA and P. GREENGARD; Raven Press, New York 1972), vol. 5, p. 151. N. TRYDING, S. E. NILSSON, G. TIFVESSON, B. BERG, S. CARLSTRO.~, B. ELMFORS, and J. E. NILSSON, Scand. J. din. Lab. Invest. 23, 79 (1969).
1214
Specialia
a v e r a g e of 6 h before d e l i v e r y to a n a v e r a g e of 78 h a f t e r 1~. H o w e v e r , t h e r e was no difference b e t w e e n p l a s m a MAO a c t i v i t y in 3rd t r i m e s t e r p r e g n a n t h u m a n females a n d c o n t r o l s in a n o t h e r s t u d y 15. R e c e n t studies h a v e s u g g e s t e d tile p o s s i b i l i t y t h a t t h e t y r a m i n e - d e a m i n a t i n g e n z y m e a c t i v i t i e s m e a s u r e d in t h e l a t t e r s t u d y a f t e r c e n t r i f u g a t i o n a t 1000-2000 g reflected r e s i d u a l p a r t i c u l a t e M A O act i v i t y f r o m plateletsl6,17. C o n s i s t e n t w i t h t h i s interpretation, the plasma MAO changes previously reported in a s s o c i a t i o n w i t h t h e m e n s t r u a l cycle in w o m e n 15,1s were n o t f o u n d b y BELMAKER et al. ~9 in p l a s m a e n z y m e t h a t h a d b e e n c e n t r i f u g e d a t 80,000 g to r e m o v e r e s i d u a l p a r t i c u l a t e e n z y m e a c t i v i t y . C h a n g e s were n o t e d i n s t e a d i n t h e p l a t e l e t MAO. W e h a v e r e p l i c a t e d t h e s e f i n d i n g s in t h e female r h e s u s m o n k e y d u r i n g t h e m e n s t r u a l cycle 3% T h e p r e s e n t s t u d y was u n d e r t a k e n t o i n v e s t i g a t e t h e possible effects of p r e g n a n c y o n p l a s m a M A O a c t i v i t y in t h e r h e s u s m o n k e y , u s i n g h i g h speed c e n t r i f u g a t i o n a n d specific i n h i b i t o r s to r e m o v e possible c o n t a m i n a t i n g m i t o chondrial enzyme. Materials and methods. 10 h e a l t h y p r e g n a n t a d u l t female r h e s u s m o n k e y s (Macaca mulatta) were s t u d i e d f r o m t h e Caribbean Primate Research Center at La Parguera. Their m e a n c e n s u s - d e t e r m i n e d age was 9.1 years ( • 1.8 SEM). A n e q u a l n u m b e r of n o n - p r e g n a n t females were s t u d i e d as controls, aged 8.7 y e a r s ( :L 1.6 SEM). 16 of t h e s e a n i m a l s were enclosed in 1/2 or 1/4 acre corrals w h e r e t h e y were fed P u r i n a m o n k e y c h o w a n d fresh w a t e r a t a single feeder a d l i b i t u m . 4 c o n t r o l m o n k e y s were i n d i v i d u a l l y caged. M o n k e y s were c a u g h t in n e t s a n d h a n d r e s t r a i n e d w i t h o u t a n e s t h e t i c s while 10 c m ~ blood was d r a w n v i a a s a p h e n o u s v e n i p u n c t u r e i n t o s o d i u m h e p a r i n i z e d syringes. T h e s p e c i m e n s were d r a w n on e a c h a n i m a l d u r i n g t h e l a t t e r p a r t of p r e g n a n c y , a f t e r delivery, or a t c o r r e s p o n d ing t i m e s of t h e y e a r for controls, a n a v e r a g e of a p p r o x i m a t e l y 4 m o n t h s a p a r t . S p e c i m e n s were p l a c e d in acid c i t r a t e d e x t r o s e (ACD) p r e s e r v a t i v e a n d cooled in a n ice b a t h p r i o r to p r e p a r a t i o n . T h e s p e c i m e n s were c e n t r i f u g e d a t 2000 g for 3 0 + m i n a t 4~ a n d p l a s m a was r e m o v e d a n d frozen o n d r y ice w i t h i n 3 h of drawing. P l a s m a was s t o r e d a t - 2 0 ~ u n t i l analysis. P l a s m a M A O was determ i n e d u s i n g a m o d i f i c a t i o n of t h e m e t h o d d e s c r i b e d b y ROBINSON e t al. ~1 u s i n g 14C-benzylamine (10 -4 M) as s u b s t r a t e . V a l u e s are r e p o r t e d as n a n o m o l e s of d e a m i n a t ed p r o d u c t / m l p l a s m a / h . A t w o - f a c t o r a n a l y s i s of v a r i a n c e w i t h r e p e a t e d m e a s u r e s on one f a c t o r was used to a n a l y z e t h e d a t a , a f t e r t e s t s of t h e h o m o g e n i e t y of v a r i a n c e s in t h e p a r t i t i o n e d e r r o r t e r m s . T e s t s of simple m a i n effects completed the analysis after a significant AB interaction was f o u n d 23 Results. P l a s m a a m i n e oxidase a c t i v i t y i n c r e a s e d in e a c h of t h e p r e g n a n t a n i m a l s f r o m t h e s p e c i m e n d u r i n g t h e p r e g n a n c y to t h e p o s t d e l i v e r y s p e c i m e n ( F = 19.1, p < 0.001). T h e r e p e a t e d m e a s u r e was n o t s i g n i f i c a n t l y d i f f e r e n t in t h e c o n t r o l s ( F = 0.2, N.S.). T h e p r e p a r t u l n m e a n was n o t s i g n i f i c a n t l y d i f f e r e n t f r o m t h e c o r r e s p o n d ing t i m e s p e c i m e n for t h e c o n t r o l s ( F -- 2.3, N.S.), while t h e p o s t - p a r t u m m e a n was s i g n i f i c a n t l y d i f f e r e n t ( F 130, p < 0.001) (Table). Discussion. T h e differences in t h i s s t u d y are b e t w e e n p o s t p a r t u m MAO a c t i v i t i e s a n d c o r r e s p o n d i n g a c t i v i t i e s f r o m n o r m a l n o n - p r e g n a n t females w i t h o u t i n f a n t s in t h a t b i r t h season, a n d b e t w e e n p o s t p a r t u m v a l u e s a n d t h e v a l u e s f r o m p r e v i o u s s p e c i m e n s d u r i n g p r e g n a n c y in t h e s a m e animals. H o r m o n a l c h a n g e s in t h e p o s t - p a r t u m p e r i o d or associated w i t h n u r s i n g i n f a n t s m a y be i n v o l v e d in t h e s e differences. Some s o c i a l l y - i n d u c e d stress r e l a t e d to b e i n g t h e m o t h e r of a y o u n g i n f a n t in a p r i m a t e social g r o u p m i g h t also be i m p l i c a t e d , a l t h o u g h we k n o w of no b e h a v i o r a l data. to s u p p o r t t h i s possibility.
EXPERIENTIA 32/9
T h e r e is e v i d e n c e f r o m o t h e r s t u d i e s t h a t m i t o c h o n d r i a l MAO a c t i v i t y in a v a r i e t y of tissues is affected b y t h e e s t r u s cycle38 or b y p r e - t r e a t m e n t w i t h estradiol or progesterone ~*. W e h a v e n o t e d effects of t h e m e n s t r u a l cycle a n d of o v a r i e c t o m y on p l a t e l e t M A O in rhesus m o n k e y s , using t y r a m i n e as s u b s t r a t e , b u t f o u n d no c h a n g e s in p l a s m a a m i n e oxidase, u s i n g benzylamineS~ T h e r e s u l t s r e p o r t e d here a p p e a r to s u p p o r t t h e f i n d i n g of n o difference in p l a s m a MAO a c t i v i t y , m e a s u r e d w i t h t y r a m i n e as s u b s t r a t e , in p r e g n a n t h u m a n females a n d c o n t r o l s b y BRIGGS a n d BRIGGS 15. O u r r e s u l t s are n o t c o n s i s t e n t w i t h t h e findings of TRYDING et al. la w h o f o u n d r e d u c e d p l a s m a MAO a c t i v i t y in h u m a n females t a k i n g s y n t h e t i c prog e s t i n - c o n t a i n i n g p h a r m a c o l o g i c agents, w h e t h e r a m i n e oxidase a c t i v i t y was m e a s u r e d w i t h b e n z y l a m i n e or t y r a m i n e as s u b s t r a t e . T h e s m a l l differences in t h e op-
P l a s m a MAO a c t i v i t y (nmoles of b e n z y l a m i n e o x i d i z e d / m l p l a s m a / h ) in p a i r e d s p e c i m e n s f r o m 10 female r h e s u s m o n k e y s a n d c o r r e s p o n d ing specimens f r o m n o n - p r e g n a n t c o n t r o l s Monkey No.
Monkey No.
Mothers Pre-
Post-partum
095 231 R65 060 0Y6 0Z1 143 685 0X3 834
7.1 10.4 23.1 39.5 17.8 13.0 67.2 34.3 11.7 9.8
16.7 17.3 31.4 121.3 50.8 19.3 81.5 85.5 25.3 45.7
Mean S.D. S;E.M.
23.4 18.8 6.0
49.5 35.6 11.3
303 075 288 08D 09G 09C 0L9 283 309 241
Controls Spec. 1
Spec. 2
25.5 13.0 5.8 21.4 48.2 24.7 22.9 36.0 43.1 27.7
23.3 21.5 13.9 31.9 29.4 24.3 15.0 30.8 28.8 18.7
26.8 12.9 4.1
23.8 6.5 2.0
Mean No. days from Delivery =82.3 +30.7 S.E.M. 8.4 7.9
1, N. J. GILMORE, D. S. IROBINSON, A. NIES, D. SYLVESTER a n d C. J . RAVARIS, J. P s y e h o s o m . Res. 15, 215 (1971). 15 M. BRIGGS a n d M. BRIGGS, J . IReprod. F e r t . 29, 447 (1972). 16 E. A. ZELLER, B. H . BABU, M. J. CAVANAUGFI a n d G. H. STANICH, P h a r m a c . ires. C o m m u n . 7, 20 (1969). 17 D. L. MURPHY a n d C. H. DONNELLY, in Neuropsychopharmacology o/ Monoamines and their Regulatory Enzymes ( R a v e n Press, N e w Y o r k 1974), p. 71. 18 E. g . KLAIBER, Y. ]5~OBAYASHI,D. M. J~ROVERMAN a n d F. HALL, J. clin. E n d o c r i n o l o g y 33, 630 (1971). 19 IR. H . ]~BLMAKER, D. L. MURPHY, IR. J . WYATT a n d D. L. L o RIAUX, A r c h . gen. P s y c h . 37, 553 (1974). 20 D. E. REDMOND, JR., D. L. MURPHY, J. BAULU, M. G. ZEIGLER a n d C. R. LAKE, P s y c h o s o m . Med. 37, 417 (1975). 21 D. S. ROBINSON, W. LOVENBERG, H . KEISER a n d J . 8JOERDSMA, Bioehem. P h a r m a e . 77, 109 (1968).
33 B. J. WINER, Statistical Principles in Experimental Design, 2nd edn. McGraw Hill, New York 1962), p. 514. 33 A. J. ZOLOVICK, IR. PEARSE, K. W. BOEHLKE and B. E. ELEFTHERIOU, Science 754, 649 (1966). 34 G. C. S. COLLINS, M. SANDLER, E. D. WILLIAMS, a n d M. B. H . YOUDIM, N a t u r e , L o u d . 225, 817 (1970).
15.9. 1976
25 H. THOMS, in Classical Contributions to Obstetrics and Gynecology (Charles C. Thomas, Springfield 1935). 26 B. C. STEVENS, Soc. Set. Med. 5, 527 (1971). 27 C. PROTHI~ROE, Br. J. Psychiat. 115, 9 (1969). 2s G. C. ROSENWALD and M. W. STONEHILL, Psyehosom, Med. 3d, 129 (1972). 29 K. DALTON, Proc. R. Soc. Med. 6d, 1249 (1971). a0 W. A. BRown and P. SHERESItEFSKY, Psychiatry 3d, 139 (1972). 31 A. NIES, D. S. ROBINSON, C. L. RAVARIS and J. M. DAvis, Psychosom. Med. 33, 470 (1971). 32 D. L. MURPHY and R. WEiss, Am. J. Psychiatr. 128, 1351 (1972). 33 j . BOCKAR, R. H. ROTH and G. HENINGER, Life Sci. 15,2109 (1974). 34 D. L. MURPhy and R. J. WYATT, Nature, Lond. 238, 225 (1972). 35 R. J. WYATT, ~). L. MURPHY, R. BELMAKER, S. COHEN, C. H. DONNEZLY and W. POLLIN, Science 179, 916 (1973). 3s A. NIES, D. S. ROmNSON, L. S. HARRIS and K. R. LAMBORN,Adv. Biochem. Psyehopharmac. 72, 59 (1974). 37 H. M~LTZER and S. M. STAHL, Res. Commun. Chem. Path. Pharmac. 7, 419 (1974).
Thyroid
1215
Specialia
Hormone
in Serum
p o s i t e d i r e c t i o n f o u n d i n t h e s t u d y b y GILMORE e t al. 14 w e r e f r o m s a m p l e s d r a w n f r o m 6 h b e f o r e t o 134 h a f t e r delivery, a short time period for which we have no comparable data. T h e s i g n i f i c a n c e of p l a s m a a m i n e o x i d a s e a c t i v i t y a n d i t s r e l a t i o n s h i p s t o b r a i n or s y m p a t h e t i c m o n o a m i n e f u n c t i o n is n o t k n o w n . D i f f e r e n c e s i n e n z y m e s r e l a t e d t o t h e m o n o a m i n e s y s t e m s a r e of i n t e r e s t b e c a u s e of p o s sible relevance to the behavioral changes associated with t h e p o s t p a r t u m p e r i o d in h u m a n s s u c h as t h e p u e r p e r a l p s y c h o s e s w h i c h h a v e b e e n r e p o r t e d s i n c e H i p p o c r a t e s 25. T h e r e is a d i s a g r e e m e n t a s t o w h e t h e r t h e s e p s y c h o s e s represent distinct nosological entities or whether factors associated with the puerperal period precipitate the exp r e s s i o n of e p i s o d e s of s c h i z o p h r e n i c o r m a n i c - d e p r e s s i v e p s y c h o s i s 2 6 - 3 0 , b o t h of w h i c h h a v e b e e n a s s o c i a t e d w i t h c h a n g e s in o t h e r m o n o a m i n e e n z y m e activities31-37.
of Fetal Calf and Pregnant
Cow During
the Last Trimester
of Pregnancy
V. STRB~K 1, F. TOMgiK2 a n d I. H L O ~ N E K 3
Institute o/ Experimental Endocrinology, Slovak Academy o[ Sciences, Kramdre, Vldrska 3, CS-809 36 Bratislava (Czechoslovakia), University School o[ Veterinary Medicine, Brno (Czechoslovakia), and Institute o[ Molecular Genetics, Czechoslovak Academy o] Sciences, Praha (Czechoslovakia), 2 March 1976. Summary. C o n s i d e r a b l y h i g h e r t h y r o x i n e a n d t r i i o d o t h y r o n i n e c o n c e n t r a t i o n s in s e r a of b o v i n e f e t u s e s t h a n i n m a t e r n a l s a m p l e s w e r e f o u n d d u r i n g t h e l a s t t r i m e s t e r of p r e g n a n c y . I t w a s s h o w n t h a t m a m m a l i a n f e t a l t h y r o i d is a c t i v e in utero and that negative feedback control appears to be f u n c t i o n a l b e f o r e p a r t u r i t i o n 4,~. T o k n o w m o r e a b o u t mother-fetus interrelation, we compared thyroid hormone c o n t e n t in s e r a of b o v i n e f e t u s e s a n d p r e g n a n t c o w s d u r i n g t h e l a s t t h i r d of p r e g n a n c y . I Material and methods. 23 p r e g n a n t C z e c h r e d - w h i t e c o w s in t h e 2 4 1 - 2 7 9 t h d a y of g e s t a t i o n w e r e c l i n i c a l l y and hematologically examined before experiment. They were submitted to high epidural anaesthesia with procain, t h e h e a d of f e t u s w a s e x t e r i o r i z e d , a r t e r i a c a r o t i s w a s c a n n u l a t e d a n d t h e f e t u s w a s e x a n g u i n a t e d s. S a m p l e s of maternal blood were withdrawn during surgical procedure. In maternal and fetal sera, thyroxine was determined by radioimmunological method ~with some modifications o f F6LDESS a n d t r i i o d o t h y r o n i n e w a s d e t e r m i n e d a c c o r d -
i n g t o NAUMAN a n d •AUMAN 9. T h y r o x i n e a n t i b o d y w a s p r o d u c e d a n d k i n d l y s u p p l i e d b y D r . F6LDES s, t r i i o d o t h y r o n i n e a n t i b o d y w a s t h e k i n d g i f t f r o m D r . NAUMAN o. Results and discussion. T h e r e s u l t s a r e s u m m a r i z e d i n the Table. Both thyroid hormones, thyroxine and trii o d o t h y r o n i n e a r e in a t l e a s t t w i c e a s h i g h in c o n c e n t r a t i o n in f e t a l s e r a a s in m a t e r n a l o n e s . S t u d e n t ' s t a n d p a i r e d tests are highly significant. We did not find any correlation between thyroid hormone values in fetuses and mothers, between thyroid hormone content in sera and body w e i g h t of f e t u s e s . W e also d i d n o t f i n d a n x t e n d e n c y of thyroid hormone concentrations to change according to a g e of f e t u s e s . I n f e t u s e s , u n l i k e m o t h e r s , p o s i t i v e c o r relation between thyroxine and triiodothyronine was noted. In the sheep fetus, high thyroxine secretion rate in c o m p a r i s o n t o m a t e r n a l o n e w a s d e s c r i b e d 1~ HA-
Thyroxine (Tt) and triiodothyronine (T3) concentrations in maternal and fetal sera. BW, body weight; n.s., non-significant
T 4 (nmol/1) T 3 (nmol/l)
Mothers
Fetuses
n
n
Means 4- SE
23 42.9 4- 3.3 19 0.50 4- 0.05
Correlation coefficients T 4 (mother: fetus) T a (mother: fetus)
23 109.4 4- 5.1 19 1.22 :J= 0.15
+ 0.07767 + 0.21016
Fetal T4:BW Ta:BW T4:T 3 Maternal T4:T 3
-- 0.42385
P < 0.001 < 0.01 n.s. n.s.
23 19 19 19
Means4- SE
+0.20033 +0.00243 + 0.45442
ms. n.s. < 0.05 n.s.
1 Acknowledgment. We wish to thank Miss R. FAJKOgOVs for skilled technical assistance. 2 University School of Veterinary Medicine, Brno, Czechoslovakia. 3 Institute of molecular Genetics, Czechoslovak Academy of Sciences, Praha, Czechoslovakia. G. t3. HEGSTAD and L. J. WELLS, Anat. Rec. 718, 389 (1954). 5 M. NIKITOWITCtt and E. KNOBIL, J. clin. Endocr. Metab. 75, 837 (1955). s F. ToragiK and L. CERN'~, In vitro v r 2/3, 46 (1973). 7 S. J. SIEGEL, W. F. LINE, N. S. YANG, A. KWONG and C. ]FRANK, J. olin. Endocr. Metab. 37, 526 (1973). s O. F6LDES, to be published. Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava. 9 A. NAUMANand J. NAUMAN, Polskie Arehw. Med. wewn. 50, 1295 (1973). 10 p. W. NATHAmELSZ, R. S. COMLINE, M. SILVER and A. L. THOMAS, J. Endocr. 58, 535 (1973). n A. ERENBER~ and D. A. FISCHER, in Foetal and Neonatal Physiology (Eds. R. S. CO,LINE, K. W. CRoss, G. S. DAVES and P. W. NATHAmELSZ, Cambridge University Press, London 1973), p. 508.
