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Effect of 6-hydroxydopamine on concentrations of neuropeptide Y (NPY) in rat female genital tract
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EFFECT OF 6 - H Y D R O X Y D O P A M I N E ON C O N C E N T R A T I O N S (NPY) IN RAT FEMALE G E N I T A L TRACT
OF N E U R O P E P T I D E
Y
JC YEATS, JM ALLEN, J GU, JM P O L A K and SR BLOOM, D e p a r t m e n t s of M e d i c i n e & Histochemistry, Royal P o s t g r a d u a t e M e d i c a l School, Du Cane Road, London W12 OHS The d i s t r i b u t i o n of n e u r o p e p t i d e Y (NPY) has been d e t e r m i n e d in the rat female u r o g e n i t a l tract. W i t h i n the bladder, high c o n c e n t r a t i o n s were found in the trigone r e g i o n (48.8 ± 5.2 pmol/g) c o m p a r e d to the dome (36.0 ± 2.1 pmol/g). NPY w a s i d e n t i f i e d thoughout the female genital tract; h i g h e s t c o n c e n t r a t i o n s b e i n g p r e s e n t in the vagina. In order to investigate the i n t e r a c t i o n of N P Y w i t h the s y m p a t h e t i c nervous system, rats were treated with 6 - h y d r o x y d o p a m i n e (60HDA), a d r u g w h i c h is a specific n e u r o t o x i n to a d r e n e r g i c nerves. Six female W i s t a r rats (200-250g) w e r e given 50 m g / k g i n t r a p e r i t o n e a l l y on two c o n s e c u t i v e days and third dose was given on the sixth day. T i s s u e s were o b t a i n e d on the seventh day after the initial dose. 6 - O H D A s i g n i f i c a n t l y d e p l e t e d c o n c e n t r a t i o n s of N P Y in both regions of the bladder, vagina, u t e r i n e horn and fallopian tube. In contrast, however, NPY c o n c e n t r a t i o n s were u n c h a n g e d within the cervix and uterine body. C h r o m a t o g r a p h i c analysis of the e x t r a c t of bladder in the control and 6-OHDA treated animals r e v e a l e d all the i m m u n o r e a c t i v i t y eluted in the p o s i t i o n of the p o r c i n e standard and no abnormal m o l e c u l a r forms were detected. D e p l e t i o n of N P Y in the bladder and three regions of the female genital tract suggests that this peptide is closely a s s o c i a t e d with the adrenergic system in this region. The failure to deplete NPY c o n c e n t r a t i o n s in the cervix and uterine body suggests that the p e p t i d e may in part be independent of the s y m p a t h e t i c nervous system.
POST SECRETORY PROCESSING OF GASTRIN IN THE PIG
D.M. Power, R. Dima]ine, N. Bunnet* and G.J. Dockray, Physiological Laboratory, U n i v e r s i t y of Liverpool, Liverpool, U.K. * A n i m a l Physiology, U n i v e r s i t y of Leeds, Leeds, U.K. We have investigated the occurrence of post secretory processing of gastrin in the pig by comparing forms in antral mucosa and antral venous plasma. Plasma and antral mucosa, taken 60 minutes a f t e r feeding, from 7 anaesthetized Yucatan pigs was f r a c t i o n a t e d by ion exchange (DEAE 52 cellulose) and gel f i l t r a t i o n (GS0) chromatography. Gastrin-tike i m m u n o r e a c t i v i t y was estimated with antisera specific for the C-terminus (1296), N - t e r m i n u s 0 2 9 5 ) or intact G17 (L6). On the DEAE four i m m u n o r e a c t i v e components were resolved from tissue, corresponding to the sulphated (s) and non-sulphated (ns) forms of (317 and (334. In plasma an additional two peaks, not present in tissue, were resolved. In tissue Q17 ns and s were present in a p p r o x i m a t e l y equimolar amounts and accounted for 93.3% + 1.6 (n = 7) of total C - t e r m i n a l i m m u n o r e a c t i v i t y , in plasma (317 ns and s accounted for 1/4% + /4.9 and 29.4% + /4.9 respectively of C - t e r m i n a l i m m u n o r e a c t i v i t y . Small amounts of-G34 were present in both antrum and plasma. The two plasma components which were absent in antral tissue together accounted for 38% + 3.7 of t o t a l plasma C - t e r m i n a l i m m u n o r e a c t i v i t y . Gel f i l t r a t i o n indicated that the two G17 peaks in antrum and plasma, and the two new components appearing in plasma, were heterogenous in that they contained C- and Nt e r m i n a l Q17 fragments. A t least two of the major forms of gastrin appearing in antral venous o u t f l o w are likely to be produced by cleavage of Q17 at or soon a f t e r secretion. Work is now in progress to characterise the main c i r c u l a t i n g fragments.
EFFECT OF 6 - H Y D R O X Y D O P A M I N E ON C O N C E N T R A T I O N S (NPY) IN RAT FEMALE G E N I T A L TRACT
OF N E U R O P E P T I D E
Y
JC YEATS, JM ALLEN, J GU, JM P O L A K and SR BLOOM, D e p a r t m e n t s of M e d i c i n e & Histochemistry, Royal P o s t g r a d u a t e M e d i c a l School, Du Cane Road, London W12 OHS The d i s t r i b u t i o n of n e u r o p e p t i d e Y (NPY) has been d e t e r m i n e d in the rat female u r o g e n i t a l tract. W i t h i n the bladder, high c o n c e n t r a t i o n s were found in the trigone r e g i o n (48.8 ± 5.2 pmol/g) c o m p a r e d to the dome (36.0 ± 2.1 pmol/g). NPY w a s i d e n t i f i e d thoughout the female genital tract; h i g h e s t c o n c e n t r a t i o n s b e i n g p r e s e n t in the vagina. In order to investigate the i n t e r a c t i o n of N P Y w i t h the s y m p a t h e t i c nervous system, rats were treated with 6 - h y d r o x y d o p a m i n e (60HDA), a d r u g w h i c h is a specific n e u r o t o x i n to a d r e n e r g i c nerves. Six female W i s t a r rats (200-250g) w e r e given 50 m g / k g i n t r a p e r i t o n e a l l y on two c o n s e c u t i v e days and third dose was given on the sixth day. T i s s u e s were o b t a i n e d on the seventh day after the initial dose. 6 - O H D A s i g n i f i c a n t l y d e p l e t e d c o n c e n t r a t i o n s of N P Y in both regions of the bladder, vagina, u t e r i n e horn and fallopian tube. In contrast, however, NPY c o n c e n t r a t i o n s were u n c h a n g e d within the cervix and uterine body. C h r o m a t o g r a p h i c analysis of the e x t r a c t of bladder in the control and 6-OHDA treated animals r e v e a l e d all the i m m u n o r e a c t i v i t y eluted in the p o s i t i o n of the p o r c i n e standard and no abnormal m o l e c u l a r forms were detected. D e p l e t i o n of N P Y in the bladder and three regions of the female genital tract suggests that this peptide is closely a s s o c i a t e d with the adrenergic system in this region. The failure to deplete NPY c o n c e n t r a t i o n s in the cervix and uterine body suggests that the p e p t i d e may in part be independent of the s y m p a t h e t i c nervous system.
POST SECRETORY PROCESSING OF GASTRIN IN THE PIG
D.M. Power, R. Dima]ine, N. Bunnet* and G.J. Dockray, Physiological Laboratory, U n i v e r s i t y of Liverpool, Liverpool, U.K. * A n i m a l Physiology, U n i v e r s i t y of Leeds, Leeds, U.K. We have investigated the occurrence of post secretory processing of gastrin in the pig by comparing forms in antral mucosa and antral venous plasma. Plasma and antral mucosa, taken 60 minutes a f t e r feeding, from 7 anaesthetized Yucatan pigs was f r a c t i o n a t e d by ion exchange (DEAE 52 cellulose) and gel f i l t r a t i o n (GS0) chromatography. Gastrin-tike i m m u n o r e a c t i v i t y was estimated with antisera specific for the C-terminus (1296), N - t e r m i n u s 0 2 9 5 ) or intact G17 (L6). On the DEAE four i m m u n o r e a c t i v e components were resolved from tissue, corresponding to the sulphated (s) and non-sulphated (ns) forms of (317 and (334. In plasma an additional two peaks, not present in tissue, were resolved. In tissue Q17 ns and s were present in a p p r o x i m a t e l y equimolar amounts and accounted for 93.3% + 1.6 (n = 7) of total C - t e r m i n a l i m m u n o r e a c t i v i t y , in plasma (317 ns and s accounted for 1/4% + /4.9 and 29.4% + /4.9 respectively of C - t e r m i n a l i m m u n o r e a c t i v i t y . Small amounts of-G34 were present in both antrum and plasma. The two plasma components which were absent in antral tissue together accounted for 38% + 3.7 of t o t a l plasma C - t e r m i n a l i m m u n o r e a c t i v i t y . Gel f i l t r a t i o n indicated that the two G17 peaks in antrum and plasma, and the two new components appearing in plasma, were heterogenous in that they contained C- and Nt e r m i n a l Q17 fragments. A t least two of the major forms of gastrin appearing in antral venous o u t f l o w are likely to be produced by cleavage of Q17 at or soon a f t e r secretion. Work is now in progress to characterise the main c i r c u l a t i n g fragments.