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Age-related changes in GRF, somatostatin, CRF and vasopressin in the rat hypothalasmus
S2 AGING-ASSOCIATED CHANGES IN N G F - D E P E N D E N T AND NGF-INDEPENDENT N E U R I T E S F R O M T I S S U E - C U L T U R E D S Y M P A T H E T I C G A N G L I A IN A C H E M I C A L L Y
REGENERATION OF DEFINED MEDIUM
*
JUN FUKUDA and KAZUKO KEINO , Department of P h y s i o l o g y , U n i v e r s i t y of Tokyo, B u n k y o - k u , T o k y o 113, J a p a n
Faculty
of
Medicine,
Abdominal sympathetic ganglia (SYG) w e r e d i s s e c t e d f r o m rabbits, rats, and m i c e of v a r i o u s ages (newborn, infant, young adult, adult and aged), and small p i e c e s of the SYG w e r e g r o w n in a c o m p l e t e l y c h e m i c a l l y d e f i n e d m e d i u m . Neurite r e g e n e r a t i o n f r o m the e x p l a n t s w a s c o m p a r e d b e t w e e n c u l t u r e s p e r f o r m e d w i t h and without nerve growth factor (NGF: e x t r a c t e d and purified from mouse salivary g l a n d s , as e i t h e r 7S or 2.5S f o r m ; t h e f i n a l c o n c e n t r a t i o n in t h e c u l t u r e media b e i n g e i t h e r 200 n g / m l f o r 7S N G F or 50 n g / m l f o r 2.5S N G F , r e s p e c t i v e l y ) . The l e n g t h of n e u r i t e s g r o w n in the p r e s e n c e of the NGF t e n d e d to be r e d u c e d w i t h the a g i n g of the h o s t a n i m a l s ; the n e u r i t e l e n g t h f r o m the SYG of 2 y e a r s old a n i m a l s was approximately 1/2 (rats) to 1/4 ( r a b b i t s ) of t h a t of t h e n e w b o r n animals. S o m e e x p l a n t s of the SYG r e p r e s e n t e d the r e g e n e r a t i o n of a s m a l l n u m b e r of s h o r t neurites e v e n in t h e a b s e n c e of t h e NGF. Such NGF-independent neurite growth b e c a m e m o r e d o m i n a n t in c u l t u r e s of the SYG f r o m the a g e d a n i m a l s ; m o s t of the SYG f r o m n e w b o r n and i n f a n t a n i m a l s f a i l e d to g r o w any n e u r i t e s w i t h o u t the NGF, w h i l e the SYG from adult and aged animals grew rather long neurites. In c u l t u r e s of t h e aged human SYG obtained from patients, n e u r i t e g r o w t h w a s s e e n n o t o n l y in t h e p r e s e n c e of t h e m o u s e 2.5S N G F b u t a l s o in t h e a b s e n c e of t h e N G F . T h e l e n g t h of t h e n e u r i t e s w i t h o u t t h e N G F w a s 1/2 to 1/4 of t h a t g r o w n w i t h t h e m o u s e 2.5S NGF. In c o n c l u s i o n , i) s y m p a t h e t i c n e r v e c e l l s r e d u c e d t h e i r c a p a b i l i t y of r e g e n e r a t i n g neurites in a g i n g , e v e n in t h e p r e s e n c e of t h e NGF. 2_! T h e S Y G n e r v e c e l l s gained NGF-independent n e u r i t e r e g e n e r a t i o n in the p r o c e s s of aging. 3) The N G F independent neurite growth remained, however, l e s s t h a n 1/2 of t h a t of t h e N G F dependent neurite growth. 4) T h e r e w a s no s i g n i f i c a n t difference between cultures made in t h e 2.5S N G F a n d t h o s e in t h e 7S N G F in t e r m s of n e u r i t e r e g e n e r a t i o n or of a g i n g - d e p e n d e n c y .
AGE-RELATEDCHANGES IN GRF, SOMATOSTATIN, CRF AND VASOPRESSIN IN THE RAT HYPOTHALASMUS
NOBUMITSU MORIMOTO I , FUMIO KAWAKAMI 2 , HITOSHI OKAMURA I , SHIGERU MAKINO I , MICHINORI HASEGAWA 3 and YASUHIKO IBATA I Dept. of IAnatomy and 2psychiatry, Kyoto Prefectural Univ. of Medicine, Kyoto
602
3Fuso Pharmaceutical Company, Osaka We
examined
age-related
changes
in GRF,
somatostatin,
CRF
and vasopressin
in the
hypothalamus of an old (24 months of age) sprague-Dawley male rat as compared with a young (3 months of age) one, using the P A P - i m m u n o h i s t o c h e m i c a l procedure.
Some animals of each age
group were injected intraventricularly with colchicine in order to investigate peptide positive perikarya, excluding vasopressin positive neurons. Both GRF and somatostatin are peptides which are related to growth hormone secretion in the hypophysis.
In the old rats, GRF immunoreactive
terminals were markedly reduced in the
external layer of the median eminence (EME) as compared with those of the young ones, although GRF immunoreactive perikarya in the arcuate nucleus were unchanged.
Somatostatin immuno-
reacative nerve terminals in the EME were not altered. On the other hand, both CRF and vasopressin are related to ACTH secretion, and their immunoreactive terminals in the EME were decreased in 24 month old rats.
The immunoreactivity of
vasopressin containing neurons was decreased particularly in the parvocellular region of the paraventricular nucleus (PVN) but was not altered in the supraoptic nucleus in the old rats. The immunoreactivity of CRF containing neurons was also decreased in the PVN of the old rats.
REGENERATION OF DEFINED MEDIUM
*
JUN FUKUDA and KAZUKO KEINO , Department of P h y s i o l o g y , U n i v e r s i t y of Tokyo, B u n k y o - k u , T o k y o 113, J a p a n
Faculty
of
Medicine,
Abdominal sympathetic ganglia (SYG) w e r e d i s s e c t e d f r o m rabbits, rats, and m i c e of v a r i o u s ages (newborn, infant, young adult, adult and aged), and small p i e c e s of the SYG w e r e g r o w n in a c o m p l e t e l y c h e m i c a l l y d e f i n e d m e d i u m . Neurite r e g e n e r a t i o n f r o m the e x p l a n t s w a s c o m p a r e d b e t w e e n c u l t u r e s p e r f o r m e d w i t h and without nerve growth factor (NGF: e x t r a c t e d and purified from mouse salivary g l a n d s , as e i t h e r 7S or 2.5S f o r m ; t h e f i n a l c o n c e n t r a t i o n in t h e c u l t u r e media b e i n g e i t h e r 200 n g / m l f o r 7S N G F or 50 n g / m l f o r 2.5S N G F , r e s p e c t i v e l y ) . The l e n g t h of n e u r i t e s g r o w n in the p r e s e n c e of the NGF t e n d e d to be r e d u c e d w i t h the a g i n g of the h o s t a n i m a l s ; the n e u r i t e l e n g t h f r o m the SYG of 2 y e a r s old a n i m a l s was approximately 1/2 (rats) to 1/4 ( r a b b i t s ) of t h a t of t h e n e w b o r n animals. S o m e e x p l a n t s of the SYG r e p r e s e n t e d the r e g e n e r a t i o n of a s m a l l n u m b e r of s h o r t neurites e v e n in t h e a b s e n c e of t h e NGF. Such NGF-independent neurite growth b e c a m e m o r e d o m i n a n t in c u l t u r e s of the SYG f r o m the a g e d a n i m a l s ; m o s t of the SYG f r o m n e w b o r n and i n f a n t a n i m a l s f a i l e d to g r o w any n e u r i t e s w i t h o u t the NGF, w h i l e the SYG from adult and aged animals grew rather long neurites. In c u l t u r e s of t h e aged human SYG obtained from patients, n e u r i t e g r o w t h w a s s e e n n o t o n l y in t h e p r e s e n c e of t h e m o u s e 2.5S N G F b u t a l s o in t h e a b s e n c e of t h e N G F . T h e l e n g t h of t h e n e u r i t e s w i t h o u t t h e N G F w a s 1/2 to 1/4 of t h a t g r o w n w i t h t h e m o u s e 2.5S NGF. In c o n c l u s i o n , i) s y m p a t h e t i c n e r v e c e l l s r e d u c e d t h e i r c a p a b i l i t y of r e g e n e r a t i n g neurites in a g i n g , e v e n in t h e p r e s e n c e of t h e NGF. 2_! T h e S Y G n e r v e c e l l s gained NGF-independent n e u r i t e r e g e n e r a t i o n in the p r o c e s s of aging. 3) The N G F independent neurite growth remained, however, l e s s t h a n 1/2 of t h a t of t h e N G F dependent neurite growth. 4) T h e r e w a s no s i g n i f i c a n t difference between cultures made in t h e 2.5S N G F a n d t h o s e in t h e 7S N G F in t e r m s of n e u r i t e r e g e n e r a t i o n or of a g i n g - d e p e n d e n c y .
AGE-RELATEDCHANGES IN GRF, SOMATOSTATIN, CRF AND VASOPRESSIN IN THE RAT HYPOTHALASMUS
NOBUMITSU MORIMOTO I , FUMIO KAWAKAMI 2 , HITOSHI OKAMURA I , SHIGERU MAKINO I , MICHINORI HASEGAWA 3 and YASUHIKO IBATA I Dept. of IAnatomy and 2psychiatry, Kyoto Prefectural Univ. of Medicine, Kyoto
602
3Fuso Pharmaceutical Company, Osaka We
examined
age-related
changes
in GRF,
somatostatin,
CRF
and vasopressin
in the
hypothalamus of an old (24 months of age) sprague-Dawley male rat as compared with a young (3 months of age) one, using the P A P - i m m u n o h i s t o c h e m i c a l procedure.
Some animals of each age
group were injected intraventricularly with colchicine in order to investigate peptide positive perikarya, excluding vasopressin positive neurons. Both GRF and somatostatin are peptides which are related to growth hormone secretion in the hypophysis.
In the old rats, GRF immunoreactive
terminals were markedly reduced in the
external layer of the median eminence (EME) as compared with those of the young ones, although GRF immunoreactive perikarya in the arcuate nucleus were unchanged.
Somatostatin immuno-
reacative nerve terminals in the EME were not altered. On the other hand, both CRF and vasopressin are related to ACTH secretion, and their immunoreactive terminals in the EME were decreased in 24 month old rats.
The immunoreactivity of
vasopressin containing neurons was decreased particularly in the parvocellular region of the paraventricular nucleus (PVN) but was not altered in the supraoptic nucleus in the old rats. The immunoreactivity of CRF containing neurons was also decreased in the PVN of the old rats.