15. 9. 1977
Speeialia
P r o t h o r a c i c g l a n d s : T h e p r o t h o r a c i c g l a n d s of S. l i t u r a are s e m i - t r a n s p a r e n t w h i t e a n d c o m p a c t , as r e p o r t e d b y Lee 8, w i t h a b o u t 51 t o 62 b e a d - l i k e cells in e a c h g l a n d w h i c h h a s a s m a l l f l a t t e n e d b o d y m a d e u p of a single row of 5 to 16 cells in t h e m i d d l e a n d d o u b l e or m o r e rows of cells t o w a r d s t h e e x t r e m i t i e s w h i c h b i f u r c a t e a n t e r i o r l y a n d posteriorly. 2 small-sized b r a n c h e s of a n t e r i o r ext r e m i t y b e a r 2 t o 3 t i e r s of bead-cells w h i c h t o u c h t h e
1249
p o s t e r i o r m a r g i n of t h e h e a d . T h e b r a n c h e s of t h e post e r i o r e x t r e m i t y are l a r g e r in size ; t h e l a t e r a l one r e a c h i n g t h e spiracle a n d t h e i n n e r one t e r m i n a t i n g n e a r t h e p r o t h o r a c i c ganglia. T h e p r o t h o r a c i c g l a n d s also increase in l e n g t h in each l a r v a l i n s t a r f r o m 0.702 m m in t h e 3rd i n s t a r t o 1.503 m m in t h e 6 t h i n s t a r l a r v a of S. l i t u r a , b u t t h e y do n o t r e v e a l a n y c h a n g e s in t h e i r f r o m e i t h e r before or a f t e r a m o u l t .
Cellular and 'subcellular ~ a l i z a t i o n of(3H~diethylstilbestrol in the ~ituitary, oViduct and~uterus of the female rat 1 P. J. S h e r i d a n
Department o/Anatomy, The University o/ Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio (Texas 78284 USA), 1 February 1977 Summary. F e m a l e r a t s were i n j e c t e d w i t h 3 H - d i e t h y l s t i l b e s t r o l . T h e p i t u i t a r i e s , o v i d u c t s , a n d u t e r i were r e m o v e d a n d processed for a u t o r a d i o g r a p h y . T h e n u c l e a r u p t a k e a n d r e t e n t i o n i n t h e s e tissues a p p e a r e d s i m i l a r to t h a t seen a f t e r t h e i n j e c t i o n of a H - e s t r a d i o l e x c e p t in t h e p a r s n e r v o s a of t h e p i t u i t a r y gland. Numerous studies using biochemical techniques have d e m o n s t r a t e d n u c l e a r u p t a k e a n d r e t e n t i o n of ~Hestradiol-17fl b y t h e p i t u i t a r y g l a n d 2, 8, t h e o v i d u c t a n d t h e u t e r u s 4. A u t o r a d i o g r a p h i e t e c h n i q u e s h a v e r e v e a l e d t h a t t h e n u c l e a r u p t a k e a n d r e t e n t i o n occurs in well o v e r
Figs. I and 2. Autoradiograms of the pituitary (figure 1), pars distaIis, and uterus (figure 2'), from 25-day - old female rats, killed 2 h after the injection of 3H-diethylstilbestrol. Exposure time 8 weeks. 4 p.m. Stained with methylgreen-pyronin. • 400.
6 0 % of t h e cells of t h e p a r s distalis, a s m a l l e r n u m b e r in t h e p a r s n e r v o s a , of t h e p i t u i t a r y g l a n d 6, a n d in t h e conn e c t i v e tissue cells, m u s c l e cells a n d e p i t h e l i a l ceils of t h e o v i d u c t a n d u t e r u s s. Since t h e c o m m o n l y used s y n t h e t i c estrogen, d i e t h y l s t i l b e s t r o l , h a s b e e n s h o w n to b i n d t o t h e e s t r o g e n r e c e p t o r in all of t h e s e tissues 3, 7 i t h a s b e e n g e n e r a l l y a s s u m e d t h a t t h e s a m e cells w h i c h t a k e u p estradiol-17fl also t a k e u p d i e t h y l s t i l b e s t r o l . T h e following s t u d y was c o n d u c t e d t o d e t e r m i n e w h e t h e r t h e n u c l e a r u p t a k e a n d r e t e n t i o n of t h i s c o m m o n l y used s y n t h e t i c e s t r o g e n is t h e s a m e as t h a t of its n a t u r a l c o u n t e r p a r t . Methods. 25-day- old f e m a l e r a t s (n = 5/group) were i n j e c t e d i.v. w i t h 1.0 ~xg of ~ H - d i e t h y l s t i l b e s t r o l (62.4 Ci/mM)/100 g b . w t o n l y or in c o m b i n a t i o n w i t h a 15-fold g r e a t e r dose of u n l a b e l e d estradiol- 178 o r u n l a b e l e d t e s t o s t e r o n e . T h e a n i m a l s were killed 2 h a f t e r t h e i n j e c t i o n , a n d f h e tissues p r o c e s s e d for a u t o r a d i o g r a p h y . T h e a u t 0 r a d i o g r a p h i c p r o c e d u r e .is d e s c r i b e d in d e t a i l elsew h e r e 8. Briefly, t h e p i t u i t a r i e s , o v i d u c t s a n d u t e r i were excised, p l a c e d o n b r a s s t i s s u e h o l d e r s a n d s i m u l t a n e o u s l y frozen and mounted by immersion in liquefied propane at about -180~ 4 m i c r o m e t e r s e c t i o n s were c u t a t -30~ k n i f e t e m p e r a t u r e in a W i d e - R a n g e C y r o s t a t ( H a r r i s M a n u f a c t u r i n g Co., C a m b r i d g e , M a s s a c h u s e t t s , U S A ) . T h e frozen sections were t h e n freeze-dried w i t h a c y r o p u m p for 24 h, a n d t h e n d r y m o u n t e d b y p r e s s u r e w i t h a t e f l o n s u p p o r t o n e m u l s i o n c o a t e d slides, p r e v i o u s l y s t o r e d o v e r drierite. T h e slides were e x p o s e d a t - 1 5 ~ for 8 weeks, t h e n d e v e l o p e d for 45 sec a t 18~ in K o d a k D-19 d e v e l o p e r , rinsed, a n d fixed for 5 m i n i n K o d a k fixer, a n d f i n a l l y rinsed a n d s t a i n e d w i t h m e t h y l g r e e n - p y r o n i n . Results. N u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y a f t e r t h e i n j e c t i o n of ~ H - d i e t h y l s t i l b e s t r o l w a s f o u n d i n well o v e r 7 0 % of t h e cells i n t h e p a r s distalis, (figure 1) b u t n o t in e i t h e r t h e p a r s i n t e r m e d i a or p a r s n e r v o s a of t h e p i t u i t a r y gland. N u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y was f o u n d in t h e c o n n e c t i v e tissue cells, m u s c l e cells, a n d epitheIiM 1 This study was supported by US PHS Career Development Award No. NS00164 and PHS Grant No. NS21933. 2 A . J . Eisenfeld, Endocrinology 86, 1313 (1970). 3 K.S. Korach and T. G. Muldoon, Biochemistry 13, 1932 (1974). 4 E.V. Jensen and E. R. DeSombre, Science 182, 126 (1973). 5 W . E . Stumpf, Z. Zellforsch., Abst. Histoehem. 92, 23 (1968). 6 W.E. Stumpf, Endocrinology 85, 31 (1969). 7 B.W. O'Malley and A. R. Means, Science 183, 610 (1974). 8 W.E. Stumpf, Acta endocr. Suppl. 153, 205 (1971).
1250
Specialia
cells of b o t h t h e o v i d u c t a n d u t e r u s (figure 2). Simult a n e o u s a d m i n i s t r a t i o n of a 15-fold g r e a t e r dose of u n labeled estradiol-17fl c o m p l e t e l y i n h i b i t e d t h e n u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y in all ceils of all 3 tissues. O n t h e o t h e r h a n d , s i m u l t a n e o u s a d m i n i s t r a t i o n of a 15-fold g r e a t e r dose of u n l a b e l e d t e s t o s t e r o n e h a d no effect o n t h e n u c l e a r c o n c e n t r a t i o n of a H - d i e t h y l s t i l b e s t r o l in t h e 3 tissues. Discussion. T h e p a t t e r n of labeled cells in t h e p i t u i t a r y agrees well w i t h t h a t r e p o r t e d a f t e r e s t r a d i o l i n j e c t i o n e x c e p t for t h e p a r s n e r v o s a ~. I n c o n t r a s t to t h e a d u l t f e m a l e r a t 5, w h e r e a n u m b e r of cells in t h e p a r s n e r v o s a were f o u n d t o be labeled a f t e r t h e i n j e c t i o n of 3H-estradiolo17fl, n o labeled cells were f o u n d in t h e p a r s n e r v o s a a f t e r t h e i n j e c t i o n of a H - d i e t h y l s t i l b e s t r o l . T h e s e d a t a are in a g r e e m e n t w i t h t h e lack of n u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y in t h e p a r s n e r v o s a in t h e 2-day-old f e m a l e r a t a f t e r t h e i n j e c t i o n of 3H-estradiol-17flg. T h e
EXPERIENTIA 33/9
n u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y in t h e o v i d u c t a n d u t e r u s a f t e r t h e i n j e c t i o n of 3 H - d i e t h y l s t i l b e s t r o l is similar t o t h a t r e p o r t e d for 3H-estradiol-17fl. T h u s , w i t h t h e e x c e p t i o n of t h e p a r s n e r v o s a of t h e p i t u i t a r y gland, t h e l o c a l i z a t i o n seen a f t e r t h e i n j e c t i o n of 3 H - d i e t h y l s t i l b e s t r o l a p p e a r s s i m i l a r to t h a t f o u n d a f t e r t h e i n j e c t i o n of 3H-estradiol-17ft. T h e difference f o u n d in t h e p a r s n e r v o s a is m o s t likely d u e t o 1 of 3 factors. First, i t m a y b e d u e to t h e s t r u c t u r a l differences b e t w e e n t h e 2 estrogens. A second p o s s i b i l i t y is t h a t t h e r e are d e v e l o p m e n t a l differences b e t w e e n t h e a d u l t a n d i m m a t u r e a n i m a l , p e r h a p s in t h e h o r m o n a l milieu. F i n a l l y , t h e differences m a y b e d u e to p r o c e d u r a l differences s u c h as v a r i a t i o n in dose or e x p o s u r e t i m e .
9
P . J . Sheridan, M. Sar and W. E. Stumpf, Experientia 29, 1415 (1973).
Section des nerfs paracardiaques internes et lib6ration provoqu6e du facteur gonadotrope m6dian chez le Criquet migrateur Section of the n e r v i corporis cardiaci I and provoked release of median gonadotropic factor in the African locust J. G i r a r d i e e t A. G i r a r d i e
Laboratoire de Zoologie, Universitd de Provence, Centre Saint-Charles, 3 Place Victor Hugo, F-13331 Marseille Cedex 3 (France), 31 January 1977 Summary. T h e in v i v o e l e c t r o s t i m u l a t i o n of t h e p a r s i n t e r c e r e b r a l i s of L o c u s t a m i g r a t o r i a , a f t e r c u t t i n g t h e n e r v i corporis c a r d i a c i I, does n o t p r o v o k e t h e release of tile g o n a d o t r o p i c h o r m o n e f r o m t h e cell bodies. T h e p r o v o k e d release of t h e g o n a d o t r o p i c h o r m o n e r e c o m m e n c e s soon as t h e a x o n t e r m i n a l s are r e g e n e r a t e d ( r e e n t r y of t h e n e r v e s i n t o t h e o r i g i n a l c o r p o r a c a r d i a c a for all tile h i s t o l o g i c a l controls). Tile e l e c t r o s t i m u l a t i o n of t h e p a r s i n t e r c e r e b r a l i s seems t o f a v o u r t h e b i l a t e r a l r e e n t r y of t h e n e r v e s i n t o t h e c o r p o r a c a r d i a c a . Les cellules n e u r o s 6 c r 6 t r i c e s p r o t o c 6 r 6 b r a l e s m 6 d i a n e s de Locusta migratoria produisent un facteur gonadotropel. L a section des nerfs p a r a c a r d i a q u e s i n t e r n e s isole les p 6 r i c a r y o n e s n e u r o s ~ c r 6 t e u r s m 6 d i a n s (group6s d a n s la p a r s i n t e r c e r e b r a l i s ) des e x t r 6 m i t 6 s a x o n i q u e s ( f o r m a n t la p a t t i e neuros6cr~trice des c o r p o r a cardiaca). C e t t e s e c t i o n e m p 6 c h e la m a t u r a t i o n o v a r i e n n e d u c r i q u e t q u a n d elle n ' e s t p a s suivie de la r 6 g 6 n 6 r a t i o n des nerfs 3. L ' e f f e t p h y s i o l o g i q u e de la l i b 6 r a t i o n d u f a c t e u r g o n a d o t r o p e n 6 c e s s i t e r a i t d o n c l ' i n t 6 g r i t 6 des cellules neuros6cr6trices m 6 d i a n e s . Ce r 6 s u l t a t est c o n f o r m e a u c o n c e p t a c t u e l l e m e n t a d m i s de la l i b 6 r a t i o n d u p r o d u i t de n e u r o s 6 c r 6 t i o n a u x e x t r 6 m i t 6 s a x o n i q u e s p o u r les cellules neuros6cr6trices d u s y s t ~ m e n e r v e u x c e n t r a l 3. C e p e n d a n t , u n e l i b 6 r a t i o n du p r o d u i t de n e u r o s 6 c r ~ t i o n p a r t i r des c o r p s cellulaires a 6t6 a v a n c 6 e p a r de n o m b r e u x histologistes a v a n t l ' a p p a r i t i o n de la microscopie 61ectronique (voir T a k e d a * ) . U n e telle l i b 6 r a t i o n s e r a i t r e s p o n s a b l e de la r u p t u r e de la d i a p a u s e chez M o n e m a flavescens e t v i e n d r a i t s ' a j o u t e r ~ l a l i b 6 r a t i o n a x o n a l e ~. Chez R h o d n i u s , le g a n g l i o n m 6 s o t h o r a c i q u e r e n f e r m e les p 6 r i c a r y o n e s de cellules neuros6cr6trices d i u r 6 t i q u e s . Sous l ' i n f l u e n c e d u p o t a s s i u m , ce g a n g l i o n lib~re in v i t r o d'importantes quantit6s d'hormones diur6tiques 5 m6me apr~s a b l a t i o n des n e r f s a b d o m i n a u x q u i c o n t i e n n e n t les e x t r 6 m i t 6 s a x o n i q u e s des cellules neuros6cr6trices. I1 n o u s a p a r u alors i n t 6 r e s s a n t d ' e x a m i n e r si les p6ric a r y o n e s n e u r o s 6 c r 6 t e u r s m 6 d i a n s de L o c u s t a , s6par6s des e x t r 6 m i t 6 s a x o n i q u e s , 6 t a l e n t c a p a b l e s de lib6rer
in v i v o leur f a c t e u r g o n a d o t r o p e sous 1'influence de chocs 61ectriques. N o u s a v o n s doric compar6, apr~s p s e u d o 6 1 e c t r o s t i m u l a t i o n (t6moins) e t ~ l e c t r o s t i m u l a t i o n (stimul6s) de la p a r s i n t e r c e r e b r a l i s e, les indices o v a r i e n s ( l o n g u e u r m o y e n n e de 10 o v o c y t e s b a s a u x ) de 99 c r i q u e t s femelles a d u l t e s r 6 p a r t i s e n 8 s6ries: 4 s6ries de t 6 m o i n s e t 4 s6ries de stimul6s. T o u s l e s c r i q u e t s o n t subi a u j o u r 1 de leur vie i m a g i n a l e u n e o u v e r t u r e collaire suivie chez 6 s6ries s e u l e m e n t de la section b i l a t 6 r a l e des nerfs p a r a c a r d i a q u e s i n t e r n e s . L a p s e u d o - 6 1 e c t r o s t i m u l a t i o n et l ' 6 1 e c t r o s t i m u l a t i o n o n t 6t6 effectu6es 1 ]our (chez les 2 s6ries g nerfs i n t a c t s e t chez 2 s6ries A nerfs sectionn6s), 4 j o u r s (chez 2 a u t r e s s6ries A nerfs sectionn6s) et 6 j o u r s (chez les 2 derni~res s6ries ~ nerfs sectionn6s) apr~s l ' o u v e r t u r e collaire. Les indices o v a r i e n s o n t 6t6 mesur6s 6 j o u r s apr~s la s t i m u l a t i o n chez les 4 s6ries h s t i m u l a t i o n pr6coce e t 4 j o u r s apr~s la s t i m u l a t i o n chez les 4 a u t r e s
1 2 3 4 5 6
A. Girardie, Bull. Soc. zool. Fr. 97, 423 (1966). M. Cazal, A. Girardie et F. Bentz, Arch. Zool. exp. g6n. 112, 293 (1971). T.C. Normann, Int. Rev. Cytol. 46, 1 (1977). N. Takeda, Appl. Ent. Zool. 11, 143 (1976). S . H . P . Maddrell et J. D. Gee, J. exp. Biol. 61,155 (1974). A. Girardie, M. Moulins et J. Girardie, J. Insect Physiol. 20, 2261 (1974).
Speeialia
P r o t h o r a c i c g l a n d s : T h e p r o t h o r a c i c g l a n d s of S. l i t u r a are s e m i - t r a n s p a r e n t w h i t e a n d c o m p a c t , as r e p o r t e d b y Lee 8, w i t h a b o u t 51 t o 62 b e a d - l i k e cells in e a c h g l a n d w h i c h h a s a s m a l l f l a t t e n e d b o d y m a d e u p of a single row of 5 to 16 cells in t h e m i d d l e a n d d o u b l e or m o r e rows of cells t o w a r d s t h e e x t r e m i t i e s w h i c h b i f u r c a t e a n t e r i o r l y a n d posteriorly. 2 small-sized b r a n c h e s of a n t e r i o r ext r e m i t y b e a r 2 t o 3 t i e r s of bead-cells w h i c h t o u c h t h e
1249
p o s t e r i o r m a r g i n of t h e h e a d . T h e b r a n c h e s of t h e post e r i o r e x t r e m i t y are l a r g e r in size ; t h e l a t e r a l one r e a c h i n g t h e spiracle a n d t h e i n n e r one t e r m i n a t i n g n e a r t h e p r o t h o r a c i c ganglia. T h e p r o t h o r a c i c g l a n d s also increase in l e n g t h in each l a r v a l i n s t a r f r o m 0.702 m m in t h e 3rd i n s t a r t o 1.503 m m in t h e 6 t h i n s t a r l a r v a of S. l i t u r a , b u t t h e y do n o t r e v e a l a n y c h a n g e s in t h e i r f r o m e i t h e r before or a f t e r a m o u l t .
Cellular and 'subcellular ~ a l i z a t i o n of(3H~diethylstilbestrol in the ~ituitary, oViduct and~uterus of the female rat 1 P. J. S h e r i d a n
Department o/Anatomy, The University o/ Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio (Texas 78284 USA), 1 February 1977 Summary. F e m a l e r a t s were i n j e c t e d w i t h 3 H - d i e t h y l s t i l b e s t r o l . T h e p i t u i t a r i e s , o v i d u c t s , a n d u t e r i were r e m o v e d a n d processed for a u t o r a d i o g r a p h y . T h e n u c l e a r u p t a k e a n d r e t e n t i o n i n t h e s e tissues a p p e a r e d s i m i l a r to t h a t seen a f t e r t h e i n j e c t i o n of a H - e s t r a d i o l e x c e p t in t h e p a r s n e r v o s a of t h e p i t u i t a r y gland. Numerous studies using biochemical techniques have d e m o n s t r a t e d n u c l e a r u p t a k e a n d r e t e n t i o n of ~Hestradiol-17fl b y t h e p i t u i t a r y g l a n d 2, 8, t h e o v i d u c t a n d t h e u t e r u s 4. A u t o r a d i o g r a p h i e t e c h n i q u e s h a v e r e v e a l e d t h a t t h e n u c l e a r u p t a k e a n d r e t e n t i o n occurs in well o v e r
Figs. I and 2. Autoradiograms of the pituitary (figure 1), pars distaIis, and uterus (figure 2'), from 25-day - old female rats, killed 2 h after the injection of 3H-diethylstilbestrol. Exposure time 8 weeks. 4 p.m. Stained with methylgreen-pyronin. • 400.
6 0 % of t h e cells of t h e p a r s distalis, a s m a l l e r n u m b e r in t h e p a r s n e r v o s a , of t h e p i t u i t a r y g l a n d 6, a n d in t h e conn e c t i v e tissue cells, m u s c l e cells a n d e p i t h e l i a l ceils of t h e o v i d u c t a n d u t e r u s s. Since t h e c o m m o n l y used s y n t h e t i c estrogen, d i e t h y l s t i l b e s t r o l , h a s b e e n s h o w n to b i n d t o t h e e s t r o g e n r e c e p t o r in all of t h e s e tissues 3, 7 i t h a s b e e n g e n e r a l l y a s s u m e d t h a t t h e s a m e cells w h i c h t a k e u p estradiol-17fl also t a k e u p d i e t h y l s t i l b e s t r o l . T h e following s t u d y was c o n d u c t e d t o d e t e r m i n e w h e t h e r t h e n u c l e a r u p t a k e a n d r e t e n t i o n of t h i s c o m m o n l y used s y n t h e t i c e s t r o g e n is t h e s a m e as t h a t of its n a t u r a l c o u n t e r p a r t . Methods. 25-day- old f e m a l e r a t s (n = 5/group) were i n j e c t e d i.v. w i t h 1.0 ~xg of ~ H - d i e t h y l s t i l b e s t r o l (62.4 Ci/mM)/100 g b . w t o n l y or in c o m b i n a t i o n w i t h a 15-fold g r e a t e r dose of u n l a b e l e d estradiol- 178 o r u n l a b e l e d t e s t o s t e r o n e . T h e a n i m a l s were killed 2 h a f t e r t h e i n j e c t i o n , a n d f h e tissues p r o c e s s e d for a u t o r a d i o g r a p h y . T h e a u t 0 r a d i o g r a p h i c p r o c e d u r e .is d e s c r i b e d in d e t a i l elsew h e r e 8. Briefly, t h e p i t u i t a r i e s , o v i d u c t s a n d u t e r i were excised, p l a c e d o n b r a s s t i s s u e h o l d e r s a n d s i m u l t a n e o u s l y frozen and mounted by immersion in liquefied propane at about -180~ 4 m i c r o m e t e r s e c t i o n s were c u t a t -30~ k n i f e t e m p e r a t u r e in a W i d e - R a n g e C y r o s t a t ( H a r r i s M a n u f a c t u r i n g Co., C a m b r i d g e , M a s s a c h u s e t t s , U S A ) . T h e frozen sections were t h e n freeze-dried w i t h a c y r o p u m p for 24 h, a n d t h e n d r y m o u n t e d b y p r e s s u r e w i t h a t e f l o n s u p p o r t o n e m u l s i o n c o a t e d slides, p r e v i o u s l y s t o r e d o v e r drierite. T h e slides were e x p o s e d a t - 1 5 ~ for 8 weeks, t h e n d e v e l o p e d for 45 sec a t 18~ in K o d a k D-19 d e v e l o p e r , rinsed, a n d fixed for 5 m i n i n K o d a k fixer, a n d f i n a l l y rinsed a n d s t a i n e d w i t h m e t h y l g r e e n - p y r o n i n . Results. N u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y a f t e r t h e i n j e c t i o n of ~ H - d i e t h y l s t i l b e s t r o l w a s f o u n d i n well o v e r 7 0 % of t h e cells i n t h e p a r s distalis, (figure 1) b u t n o t in e i t h e r t h e p a r s i n t e r m e d i a or p a r s n e r v o s a of t h e p i t u i t a r y gland. N u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y was f o u n d in t h e c o n n e c t i v e tissue cells, m u s c l e cells, a n d epitheIiM 1 This study was supported by US PHS Career Development Award No. NS00164 and PHS Grant No. NS21933. 2 A . J . Eisenfeld, Endocrinology 86, 1313 (1970). 3 K.S. Korach and T. G. Muldoon, Biochemistry 13, 1932 (1974). 4 E.V. Jensen and E. R. DeSombre, Science 182, 126 (1973). 5 W . E . Stumpf, Z. Zellforsch., Abst. Histoehem. 92, 23 (1968). 6 W.E. Stumpf, Endocrinology 85, 31 (1969). 7 B.W. O'Malley and A. R. Means, Science 183, 610 (1974). 8 W.E. Stumpf, Acta endocr. Suppl. 153, 205 (1971).
1250
Specialia
cells of b o t h t h e o v i d u c t a n d u t e r u s (figure 2). Simult a n e o u s a d m i n i s t r a t i o n of a 15-fold g r e a t e r dose of u n labeled estradiol-17fl c o m p l e t e l y i n h i b i t e d t h e n u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y in all ceils of all 3 tissues. O n t h e o t h e r h a n d , s i m u l t a n e o u s a d m i n i s t r a t i o n of a 15-fold g r e a t e r dose of u n l a b e l e d t e s t o s t e r o n e h a d no effect o n t h e n u c l e a r c o n c e n t r a t i o n of a H - d i e t h y l s t i l b e s t r o l in t h e 3 tissues. Discussion. T h e p a t t e r n of labeled cells in t h e p i t u i t a r y agrees well w i t h t h a t r e p o r t e d a f t e r e s t r a d i o l i n j e c t i o n e x c e p t for t h e p a r s n e r v o s a ~. I n c o n t r a s t to t h e a d u l t f e m a l e r a t 5, w h e r e a n u m b e r of cells in t h e p a r s n e r v o s a were f o u n d t o be labeled a f t e r t h e i n j e c t i o n of 3H-estradiolo17fl, n o labeled cells were f o u n d in t h e p a r s n e r v o s a a f t e r t h e i n j e c t i o n of a H - d i e t h y l s t i l b e s t r o l . T h e s e d a t a are in a g r e e m e n t w i t h t h e lack of n u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y in t h e p a r s n e r v o s a in t h e 2-day-old f e m a l e r a t a f t e r t h e i n j e c t i o n of 3H-estradiol-17flg. T h e
EXPERIENTIA 33/9
n u c l e a r c o n c e n t r a t i o n of r a d i o a c t i v i t y in t h e o v i d u c t a n d u t e r u s a f t e r t h e i n j e c t i o n of 3 H - d i e t h y l s t i l b e s t r o l is similar t o t h a t r e p o r t e d for 3H-estradiol-17fl. T h u s , w i t h t h e e x c e p t i o n of t h e p a r s n e r v o s a of t h e p i t u i t a r y gland, t h e l o c a l i z a t i o n seen a f t e r t h e i n j e c t i o n of 3 H - d i e t h y l s t i l b e s t r o l a p p e a r s s i m i l a r to t h a t f o u n d a f t e r t h e i n j e c t i o n of 3H-estradiol-17ft. T h e difference f o u n d in t h e p a r s n e r v o s a is m o s t likely d u e t o 1 of 3 factors. First, i t m a y b e d u e to t h e s t r u c t u r a l differences b e t w e e n t h e 2 estrogens. A second p o s s i b i l i t y is t h a t t h e r e are d e v e l o p m e n t a l differences b e t w e e n t h e a d u l t a n d i m m a t u r e a n i m a l , p e r h a p s in t h e h o r m o n a l milieu. F i n a l l y , t h e differences m a y b e d u e to p r o c e d u r a l differences s u c h as v a r i a t i o n in dose or e x p o s u r e t i m e .
9
P . J . Sheridan, M. Sar and W. E. Stumpf, Experientia 29, 1415 (1973).
Section des nerfs paracardiaques internes et lib6ration provoqu6e du facteur gonadotrope m6dian chez le Criquet migrateur Section of the n e r v i corporis cardiaci I and provoked release of median gonadotropic factor in the African locust J. G i r a r d i e e t A. G i r a r d i e
Laboratoire de Zoologie, Universitd de Provence, Centre Saint-Charles, 3 Place Victor Hugo, F-13331 Marseille Cedex 3 (France), 31 January 1977 Summary. T h e in v i v o e l e c t r o s t i m u l a t i o n of t h e p a r s i n t e r c e r e b r a l i s of L o c u s t a m i g r a t o r i a , a f t e r c u t t i n g t h e n e r v i corporis c a r d i a c i I, does n o t p r o v o k e t h e release of tile g o n a d o t r o p i c h o r m o n e f r o m t h e cell bodies. T h e p r o v o k e d release of t h e g o n a d o t r o p i c h o r m o n e r e c o m m e n c e s soon as t h e a x o n t e r m i n a l s are r e g e n e r a t e d ( r e e n t r y of t h e n e r v e s i n t o t h e o r i g i n a l c o r p o r a c a r d i a c a for all tile h i s t o l o g i c a l controls). Tile e l e c t r o s t i m u l a t i o n of t h e p a r s i n t e r c e r e b r a l i s seems t o f a v o u r t h e b i l a t e r a l r e e n t r y of t h e n e r v e s i n t o t h e c o r p o r a c a r d i a c a . Les cellules n e u r o s 6 c r 6 t r i c e s p r o t o c 6 r 6 b r a l e s m 6 d i a n e s de Locusta migratoria produisent un facteur gonadotropel. L a section des nerfs p a r a c a r d i a q u e s i n t e r n e s isole les p 6 r i c a r y o n e s n e u r o s ~ c r 6 t e u r s m 6 d i a n s (group6s d a n s la p a r s i n t e r c e r e b r a l i s ) des e x t r 6 m i t 6 s a x o n i q u e s ( f o r m a n t la p a t t i e neuros6cr~trice des c o r p o r a cardiaca). C e t t e s e c t i o n e m p 6 c h e la m a t u r a t i o n o v a r i e n n e d u c r i q u e t q u a n d elle n ' e s t p a s suivie de la r 6 g 6 n 6 r a t i o n des nerfs 3. L ' e f f e t p h y s i o l o g i q u e de la l i b 6 r a t i o n d u f a c t e u r g o n a d o t r o p e n 6 c e s s i t e r a i t d o n c l ' i n t 6 g r i t 6 des cellules neuros6cr6trices m 6 d i a n e s . Ce r 6 s u l t a t est c o n f o r m e a u c o n c e p t a c t u e l l e m e n t a d m i s de la l i b 6 r a t i o n d u p r o d u i t de n e u r o s 6 c r 6 t i o n a u x e x t r 6 m i t 6 s a x o n i q u e s p o u r les cellules neuros6cr6trices d u s y s t ~ m e n e r v e u x c e n t r a l 3. C e p e n d a n t , u n e l i b 6 r a t i o n du p r o d u i t de n e u r o s 6 c r ~ t i o n p a r t i r des c o r p s cellulaires a 6t6 a v a n c 6 e p a r de n o m b r e u x histologistes a v a n t l ' a p p a r i t i o n de la microscopie 61ectronique (voir T a k e d a * ) . U n e telle l i b 6 r a t i o n s e r a i t r e s p o n s a b l e de la r u p t u r e de la d i a p a u s e chez M o n e m a flavescens e t v i e n d r a i t s ' a j o u t e r ~ l a l i b 6 r a t i o n a x o n a l e ~. Chez R h o d n i u s , le g a n g l i o n m 6 s o t h o r a c i q u e r e n f e r m e les p 6 r i c a r y o n e s de cellules neuros6cr6trices d i u r 6 t i q u e s . Sous l ' i n f l u e n c e d u p o t a s s i u m , ce g a n g l i o n lib~re in v i t r o d'importantes quantit6s d'hormones diur6tiques 5 m6me apr~s a b l a t i o n des n e r f s a b d o m i n a u x q u i c o n t i e n n e n t les e x t r 6 m i t 6 s a x o n i q u e s des cellules neuros6cr6trices. I1 n o u s a p a r u alors i n t 6 r e s s a n t d ' e x a m i n e r si les p6ric a r y o n e s n e u r o s 6 c r 6 t e u r s m 6 d i a n s de L o c u s t a , s6par6s des e x t r 6 m i t 6 s a x o n i q u e s , 6 t a l e n t c a p a b l e s de lib6rer
in v i v o leur f a c t e u r g o n a d o t r o p e sous 1'influence de chocs 61ectriques. N o u s a v o n s doric compar6, apr~s p s e u d o 6 1 e c t r o s t i m u l a t i o n (t6moins) e t ~ l e c t r o s t i m u l a t i o n (stimul6s) de la p a r s i n t e r c e r e b r a l i s e, les indices o v a r i e n s ( l o n g u e u r m o y e n n e de 10 o v o c y t e s b a s a u x ) de 99 c r i q u e t s femelles a d u l t e s r 6 p a r t i s e n 8 s6ries: 4 s6ries de t 6 m o i n s e t 4 s6ries de stimul6s. T o u s l e s c r i q u e t s o n t subi a u j o u r 1 de leur vie i m a g i n a l e u n e o u v e r t u r e collaire suivie chez 6 s6ries s e u l e m e n t de la section b i l a t 6 r a l e des nerfs p a r a c a r d i a q u e s i n t e r n e s . L a p s e u d o - 6 1 e c t r o s t i m u l a t i o n et l ' 6 1 e c t r o s t i m u l a t i o n o n t 6t6 effectu6es 1 ]our (chez les 2 s6ries g nerfs i n t a c t s e t chez 2 s6ries A nerfs sectionn6s), 4 j o u r s (chez 2 a u t r e s s6ries A nerfs sectionn6s) et 6 j o u r s (chez les 2 derni~res s6ries ~ nerfs sectionn6s) apr~s l ' o u v e r t u r e collaire. Les indices o v a r i e n s o n t 6t6 mesur6s 6 j o u r s apr~s la s t i m u l a t i o n chez les 4 s6ries h s t i m u l a t i o n pr6coce e t 4 j o u r s apr~s la s t i m u l a t i o n chez les 4 a u t r e s
1 2 3 4 5 6
A. Girardie, Bull. Soc. zool. Fr. 97, 423 (1966). M. Cazal, A. Girardie et F. Bentz, Arch. Zool. exp. g6n. 112, 293 (1971). T.C. Normann, Int. Rev. Cytol. 46, 1 (1977). N. Takeda, Appl. Ent. Zool. 11, 143 (1976). S . H . P . Maddrell et J. D. Gee, J. exp. Biol. 61,155 (1974). A. Girardie, M. Moulins et J. Girardie, J. Insect Physiol. 20, 2261 (1974).
