- Email: [email protected]
Production of glucagon and gut glucagon-like immunoreactivity by a human pancreatic glucagonoma in culture
332
Intravenous but not intragastric urogastrone-EGF is trophic to the of parenterally fed rats
intestine
R A Goodlad, T J G Wilson, W Lenton, H Gregory *, K G McCullagh**, N A Wright. Department of Histopathology,Royal Postgraduate Medical School, Ham~ersmith Hospital, Ducane Road, London. * ICI, Alderley Park, Macclesfield. ** G D Searle, High Wycombe, Bucks. Rats were maintained on total parenteral nutrition (TPN) in order to reduce intestinal epithelial cell proliferation to a steady state basal level and to lessen the effects of luminal nutrition and endogenous secretions. The wet weight of the stomach,small intestine and colon and the mucosal crypt cell production rate (CCPR) of these tissues were significantly decreased (P<0.01 to 0.001) after ten days on an isocaloric TPN diet when compared to orally fed controls. Continuous infusion of 15 pg per rat per day of recombinant beta urogastrone, a dose below that needed to inhibit gastric acid secretion, significantly increased (P<0.01 to 0.001 ) both the weights of the various sections of the intestine and the CCPR of five sites in the intestine (P<0.05 to 0.001). Increasing doses of urogastrone progressively elevated the two hour collection of metaphases and the weights of the intestine, with the colon showing the most pronounced effects. Intravenous infusion of urogastrone was also effective in restoring cell proliferation when it was only infused after the intestine had become hypoproliferative. 1 5 ~ g per rat per day of urogastrone administered via an intragastric cannulae thrice daily had no significant effect on intestinal weight or CCPR, neither did the luminal administration of higher doses of urogasrtone (150 and 300 ~g/rat/day) have any significant effect on intestinal weight or 2 hour metaphase collection. It is proposed that one of the in vivo actions of urogastrone-EGF is the maintenance of gastrointestinal growth and that this occurs primarily via a systemic mechanism.
PRODUCl"ION OF ~UCAGON AIO 9UT ~LUCA~iq-.-LI]~ IMI~O~K&CI"IVITT BY A ~ PANCREATIC ~LUCA~0NONA IN CULT0~E C. GRAY, A. WHITE, K. TAN, I . ROBERTS AND Y.G. RATCLIFFE, D e p t . o f Chem. P a t h . , Univ. o f M a n c h e s t e r , Hope H o s p i t a l , S a l f o r d , M6 8HD and Div. o f C l i n i c a l B i o c h e m i s t r y Univ. o f S u r r e y , G u i l d f o r d , GU2 5XH A pancreatic presented
glucagon
islet
cell
tumour
was
removed
from a patient
who
with the classical g l u c a g o n o m a syndrome, i n c l u d i n g r a i s e d p l a s m a ( 1 9 2 0 n g / 1 ) and h i g h l e v e l s o f g u t g l u c a g o n - l i k e immunoreactivity
(gut-GLI, 5910ng equiv/1). At o p e r a t i o n , jejunal hypertrophy was n o t e d . Immunohistochemioal studies demonstrated 81ucagon-reactive tumour cells. For in vitro studies the tumour was dispersed with or without trypsin at 37°C using a teflon paddle. Cells were cultured in medium containing 20% f o e t a l calf serum with fibronectin or conditioned medium to aid cell survival. Morphologically the cell population consisted of granular secretory cells p r e s e n t as c l u m p s o r i n t e r s p e r s e d w i t h f i b r o b l a s t s . The c e l l s w e r e m a i n t a i n e d in c u l t u r e f o r a p e r i o d o f 63 d a y s and s e c r e t e d h i g h levels of immunoreactive (IR) p a n c r e a t i c g l u c a g o n (up t o 2 4 0 0 r i g / l ) and gUt
GLI, (up t o 8 0 , 0 0 0 n g e q u i v / l ) .
IR s o m a t o s t a t i n
(up t o 2 5 0 0 n g / 1 ) ,
IR i n s u l i n
(up t o 302 mU/l) and l o w , b u t s i g n i f i c a n t l e v e l s o f N- and C- t e r m i n a l VIP w e r e a l s o d e t e c t e d . T h e r e was no e v i d e n c e o f g a s t r i n o r ACTH s e c r e t i o n by a n y cultures. Secretion o f g l u c a g o n and s o m a t o s t a t i n persisted in several cultures f o r 63 d a y s , h u t g u t - G L I and i n s u l i n l e v e l s d e c l i n e d . T h e r e w a s no direct or inverse correlation between d i f f e r e n t hormone l e v e l s . Incubation of t u m o u r c e l l s w i t h e x o g e n o u s s o m a t o s t a t i n ( I n g / m l ) i n h i b i t e d glucagon s e c r e t i o n by 75%, w h e r e a s a d d i t i o n o f i n s u l i n up t o I 0 0 0 mU/l had no e f f e c t on g l u c a g o n s e c r e t i o n b u t p r o d u c e d a 70~ i n h i b i t i o n o f s o m a t o s t a t i n s e c r e t i o n .
Intravenous but not intragastric urogastrone-EGF is trophic to the of parenterally fed rats
intestine
R A Goodlad, T J G Wilson, W Lenton, H Gregory *, K G McCullagh**, N A Wright. Department of Histopathology,Royal Postgraduate Medical School, Ham~ersmith Hospital, Ducane Road, London. * ICI, Alderley Park, Macclesfield. ** G D Searle, High Wycombe, Bucks. Rats were maintained on total parenteral nutrition (TPN) in order to reduce intestinal epithelial cell proliferation to a steady state basal level and to lessen the effects of luminal nutrition and endogenous secretions. The wet weight of the stomach,small intestine and colon and the mucosal crypt cell production rate (CCPR) of these tissues were significantly decreased (P<0.01 to 0.001) after ten days on an isocaloric TPN diet when compared to orally fed controls. Continuous infusion of 15 pg per rat per day of recombinant beta urogastrone, a dose below that needed to inhibit gastric acid secretion, significantly increased (P<0.01 to 0.001 ) both the weights of the various sections of the intestine and the CCPR of five sites in the intestine (P<0.05 to 0.001). Increasing doses of urogastrone progressively elevated the two hour collection of metaphases and the weights of the intestine, with the colon showing the most pronounced effects. Intravenous infusion of urogastrone was also effective in restoring cell proliferation when it was only infused after the intestine had become hypoproliferative. 1 5 ~ g per rat per day of urogastrone administered via an intragastric cannulae thrice daily had no significant effect on intestinal weight or CCPR, neither did the luminal administration of higher doses of urogasrtone (150 and 300 ~g/rat/day) have any significant effect on intestinal weight or 2 hour metaphase collection. It is proposed that one of the in vivo actions of urogastrone-EGF is the maintenance of gastrointestinal growth and that this occurs primarily via a systemic mechanism.
PRODUCl"ION OF ~UCAGON AIO 9UT ~LUCA~iq-.-LI]~ IMI~O~K&CI"IVITT BY A ~ PANCREATIC ~LUCA~0NONA IN CULT0~E C. GRAY, A. WHITE, K. TAN, I . ROBERTS AND Y.G. RATCLIFFE, D e p t . o f Chem. P a t h . , Univ. o f M a n c h e s t e r , Hope H o s p i t a l , S a l f o r d , M6 8HD and Div. o f C l i n i c a l B i o c h e m i s t r y Univ. o f S u r r e y , G u i l d f o r d , GU2 5XH A pancreatic presented
glucagon
islet
cell
tumour
was
removed
from a patient
who
with the classical g l u c a g o n o m a syndrome, i n c l u d i n g r a i s e d p l a s m a ( 1 9 2 0 n g / 1 ) and h i g h l e v e l s o f g u t g l u c a g o n - l i k e immunoreactivity
(gut-GLI, 5910ng equiv/1). At o p e r a t i o n , jejunal hypertrophy was n o t e d . Immunohistochemioal studies demonstrated 81ucagon-reactive tumour cells. For in vitro studies the tumour was dispersed with or without trypsin at 37°C using a teflon paddle. Cells were cultured in medium containing 20% f o e t a l calf serum with fibronectin or conditioned medium to aid cell survival. Morphologically the cell population consisted of granular secretory cells p r e s e n t as c l u m p s o r i n t e r s p e r s e d w i t h f i b r o b l a s t s . The c e l l s w e r e m a i n t a i n e d in c u l t u r e f o r a p e r i o d o f 63 d a y s and s e c r e t e d h i g h levels of immunoreactive (IR) p a n c r e a t i c g l u c a g o n (up t o 2 4 0 0 r i g / l ) and gUt
GLI, (up t o 8 0 , 0 0 0 n g e q u i v / l ) .
IR s o m a t o s t a t i n
(up t o 2 5 0 0 n g / 1 ) ,
IR i n s u l i n
(up t o 302 mU/l) and l o w , b u t s i g n i f i c a n t l e v e l s o f N- and C- t e r m i n a l VIP w e r e a l s o d e t e c t e d . T h e r e was no e v i d e n c e o f g a s t r i n o r ACTH s e c r e t i o n by a n y cultures. Secretion o f g l u c a g o n and s o m a t o s t a t i n persisted in several cultures f o r 63 d a y s , h u t g u t - G L I and i n s u l i n l e v e l s d e c l i n e d . T h e r e w a s no direct or inverse correlation between d i f f e r e n t hormone l e v e l s . Incubation of t u m o u r c e l l s w i t h e x o g e n o u s s o m a t o s t a t i n ( I n g / m l ) i n h i b i t e d glucagon s e c r e t i o n by 75%, w h e r e a s a d d i t i o n o f i n s u l i n up t o I 0 0 0 mU/l had no e f f e c t on g l u c a g o n s e c r e t i o n b u t p r o d u c e d a 70~ i n h i b i t i o n o f s o m a t o s t a t i n s e c r e t i o n .