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Light microscopic immunohistochemistry reveals species-dependent presence of tachykinins in intrinsic neurons in the mammalian respiratory tract
425 LIGHT MICROSCOPIC IMMUNOHISTOCHEMISTRY REVEALS SPECIES-DEPENDENT PRESENCE OF TACHYKININS IN INTRINSIC NEURONS IN THE MAMMALIAN RESPIRATORY TRACT D. NOHR and E. WEIHE, A n a t o m i c a l Institute, F.R.G.
U n i v e r s i t y of Mainz,
D-6500 Mainz,
The t a c h y k i n i n s s u b s t a n c e P (SP) and n e u r o k i n i n A (NKA) a r e p r e s e n t i n c a p s a i c i n - s e n s i t i v e v a g a l s e n s o r y f i b r e s as m a i n l y i n v e s t i g a t e d i n t h e g u i n e a p i g . U n l i k e VIP, t h e y have been r e p o r t e d t o be a b s e n t from i n t r i n s i c n e u r o n s of the respiratory tract. This s t u d y i n v e s t i g a t e s t h e p o s s i b i l i t y that the r e p o r t e d a b s e n c e of t a c h y k i n i n s from i n t r i n s i c n e u r o n s may have been due t o as yet unelucidated interspecies variations. Therefore, t h e lower r e s p i r a t o r y tract of g u i n e a - p i g , r a t and c a t was p r o c e s s e d f o r enzyme and f l u o r e s c e n c e i a t a u n o h i s t o c h e m i s t r y . In a l l s p e c i e s i n v e s t i g a t e d , SP- and V I P - i m a u n o r e a c t i v e (Jr) nerve f i b r e s s u p p l i e d the r e s p i r a t o r y e p i t h e l i u m , b r o n c h i a l and t r a c h e a l b l o o d v e s s e l s and smooth m u s c l e s as w e l l as seromucous g l a n d s . A l a r g e number of p e r i k a r y a o f i n t r i n s i c g a n g l i a s t a i n e d f o r r I P . In t h e c a t , a s u b p o p u l a t i o n of i n t r i n s i c t r a c h e a l and b r o n c h i a l n e u r o n s s t a i n e d a l s o f o r SP. I n c o n t r a s t , SP s t a i n i n g was a b s e n t from such n e u r o n s o f g u i n e a - p i g and r a t a l t h o u g h i n t e n s e staining f o r SP was s e e n i n f i b r e s s u p p l y i n g i n t r i n s i c g a n g l i a of t h e s e s p e c i e s . At l e a s t some o f t h e i n t r i n s i c S P - i r n e u r o n s i n t h e c a t a l s o c o n t a i n e d VIP. There was a l s o c o e x i s t e n c e of r I P and SP i n a s u b p o p u l a t i o n o f f i b r e s . Our results c o n f i r m v e r y r e c e n t o b s e r v a t i o n s of o t h e r a u t h o r s showing t h e p r e s e n c e of SP i n i n t r i n s i c n e u r o n s o f c a t r e s p i r a t o r y t r a c t . I t w i l l be c r u c i a l to determine whether intrinsic tachykinin neurons also occur in the respiratory t r a c t of o t h e r n o n - r o d e n t s , p a r t i c u l a r l y i n t h a t of man, b e f o r e c o n c l u s i o n s can be drawn about t h e i r physiological and p a t h o p h y s i o l o g i c a l s i g n i f i c a n c e . S u p p o r t e d by t h e German R e s e a r c h F o u n d a t i o n .
THE E F F E C T O F O V A R I E C T O M Y A N D O E S T R O G E N R E P L A C E M E N T O N THE A N T E R I O R P I T U I T A R Y P E P T I D E S 7B2 A N D G A L A N I N IN THE RAT. D.J. O ' H A L L O R A N , M.A. GHATEI ,J.M. B U R R I N , J.H. S T E E L I, ~.M. P O L A K 1 and S.R. BLOOM. D e p a r t m e n t s of M e d i c i n e and H i s t o c h e m i s t r y , Royal Postgraduate Medical School,London WI20HS. 7B2 and g a l a n i n (GAL) are present in h i g h c o n c e n t r a t i o n in the rat a n t e r i o r ~ i t u i t a r y gland. We i n v e s t i g a t e d the effect of o v a r i e c t o m y on the a n t e r l o r p i t u i t a r y c o n c e n t r a t i o n of these p e p t i d e s in the adult rat. A r e p l a c e m e n t g r o u ~ r e c e i v e d o e s t r a d i o l SuE s.c. d a i l y for 28 days. A n i m a l s were s a c r l f i c e d at 42 days post-surgery. A n t e r i o r p i t u i t a r i e s w s r e e x t r a c t e d in 0.5M acetic acid for 10 m i n u t e s and stored at -20 C prior to a s s a y in duplicate. 7B2 and GAL c o n c e n t r a t i o n s were m e a s u r e d by a specific radioimmunoassay. G o n a d o t r o p h i n and o e s t r a d i o l levels proved the e f f i c a c y of the endocrine m a n i p u l a t i o n . R e s u l t s are expressed as m e a n ~ SEM (n=10). GROUP control ovariectomised replaced
GAL(fmol/21and) 185 ! 8 91 ~12 163 ~ 8
7B2fDmol/~land] 12.9 ~ 0.6 26.2 ~ 1.0 14.4 ~ 1.3
7B2 shows a s i g n i f i c a n t i n c r e a s e in female rats following ovariectomy. This effect was r e v e r s e d w i t h p h y s i o l o g i c a l o e s t r o g e n replacement. G A L shows a s i g n i f i c a n t and r e v e r s i b l e d e c r e a s e following ovariectomy. These r e s u l t s w o u l d suggest that the a n t e r i o r p i t u i t a r y c o n c e n t r a t i o n of 7B2 and GAL are o e s t r o g e n r e g u l a t e d in the female rat. The functional role of these o b s e r v a t i o n s is not understood.
U n i v e r s i t y of Mainz,
D-6500 Mainz,
The t a c h y k i n i n s s u b s t a n c e P (SP) and n e u r o k i n i n A (NKA) a r e p r e s e n t i n c a p s a i c i n - s e n s i t i v e v a g a l s e n s o r y f i b r e s as m a i n l y i n v e s t i g a t e d i n t h e g u i n e a p i g . U n l i k e VIP, t h e y have been r e p o r t e d t o be a b s e n t from i n t r i n s i c n e u r o n s of the respiratory tract. This s t u d y i n v e s t i g a t e s t h e p o s s i b i l i t y that the r e p o r t e d a b s e n c e of t a c h y k i n i n s from i n t r i n s i c n e u r o n s may have been due t o as yet unelucidated interspecies variations. Therefore, t h e lower r e s p i r a t o r y tract of g u i n e a - p i g , r a t and c a t was p r o c e s s e d f o r enzyme and f l u o r e s c e n c e i a t a u n o h i s t o c h e m i s t r y . In a l l s p e c i e s i n v e s t i g a t e d , SP- and V I P - i m a u n o r e a c t i v e (Jr) nerve f i b r e s s u p p l i e d the r e s p i r a t o r y e p i t h e l i u m , b r o n c h i a l and t r a c h e a l b l o o d v e s s e l s and smooth m u s c l e s as w e l l as seromucous g l a n d s . A l a r g e number of p e r i k a r y a o f i n t r i n s i c g a n g l i a s t a i n e d f o r r I P . In t h e c a t , a s u b p o p u l a t i o n of i n t r i n s i c t r a c h e a l and b r o n c h i a l n e u r o n s s t a i n e d a l s o f o r SP. I n c o n t r a s t , SP s t a i n i n g was a b s e n t from such n e u r o n s o f g u i n e a - p i g and r a t a l t h o u g h i n t e n s e staining f o r SP was s e e n i n f i b r e s s u p p l y i n g i n t r i n s i c g a n g l i a of t h e s e s p e c i e s . At l e a s t some o f t h e i n t r i n s i c S P - i r n e u r o n s i n t h e c a t a l s o c o n t a i n e d VIP. There was a l s o c o e x i s t e n c e of r I P and SP i n a s u b p o p u l a t i o n o f f i b r e s . Our results c o n f i r m v e r y r e c e n t o b s e r v a t i o n s of o t h e r a u t h o r s showing t h e p r e s e n c e of SP i n i n t r i n s i c n e u r o n s o f c a t r e s p i r a t o r y t r a c t . I t w i l l be c r u c i a l to determine whether intrinsic tachykinin neurons also occur in the respiratory t r a c t of o t h e r n o n - r o d e n t s , p a r t i c u l a r l y i n t h a t of man, b e f o r e c o n c l u s i o n s can be drawn about t h e i r physiological and p a t h o p h y s i o l o g i c a l s i g n i f i c a n c e . S u p p o r t e d by t h e German R e s e a r c h F o u n d a t i o n .
THE E F F E C T O F O V A R I E C T O M Y A N D O E S T R O G E N R E P L A C E M E N T O N THE A N T E R I O R P I T U I T A R Y P E P T I D E S 7B2 A N D G A L A N I N IN THE RAT. D.J. O ' H A L L O R A N , M.A. GHATEI ,J.M. B U R R I N , J.H. S T E E L I, ~.M. P O L A K 1 and S.R. BLOOM. D e p a r t m e n t s of M e d i c i n e and H i s t o c h e m i s t r y , Royal Postgraduate Medical School,London WI20HS. 7B2 and g a l a n i n (GAL) are present in h i g h c o n c e n t r a t i o n in the rat a n t e r i o r ~ i t u i t a r y gland. We i n v e s t i g a t e d the effect of o v a r i e c t o m y on the a n t e r l o r p i t u i t a r y c o n c e n t r a t i o n of these p e p t i d e s in the adult rat. A r e p l a c e m e n t g r o u ~ r e c e i v e d o e s t r a d i o l SuE s.c. d a i l y for 28 days. A n i m a l s were s a c r l f i c e d at 42 days post-surgery. A n t e r i o r p i t u i t a r i e s w s r e e x t r a c t e d in 0.5M acetic acid for 10 m i n u t e s and stored at -20 C prior to a s s a y in duplicate. 7B2 and GAL c o n c e n t r a t i o n s were m e a s u r e d by a specific radioimmunoassay. G o n a d o t r o p h i n and o e s t r a d i o l levels proved the e f f i c a c y of the endocrine m a n i p u l a t i o n . R e s u l t s are expressed as m e a n ~ SEM (n=10). GROUP control ovariectomised replaced
GAL(fmol/21and) 185 ! 8 91 ~12 163 ~ 8
7B2fDmol/~land] 12.9 ~ 0.6 26.2 ~ 1.0 14.4 ~ 1.3
7B2 shows a s i g n i f i c a n t i n c r e a s e in female rats following ovariectomy. This effect was r e v e r s e d w i t h p h y s i o l o g i c a l o e s t r o g e n replacement. G A L shows a s i g n i f i c a n t and r e v e r s i b l e d e c r e a s e following ovariectomy. These r e s u l t s w o u l d suggest that the a n t e r i o r p i t u i t a r y c o n c e n t r a t i o n of 7B2 and GAL are o e s t r o g e n r e g u l a t e d in the female rat. The functional role of these o b s e r v a t i o n s is not understood.