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Cytokines in the reproduction of lower vertebrates
A.58
Placenta (1993), I/ol 14
E F F E C T S OF A L K A L I N E P H O S P H A T A S E I N H I B I T O R S U P O N R A D I O C A L C I U M UPTAKE BY M I C R O V I L L O U S BORDER MEMBRANES OF THE HUMAN P L A C E N T A . K . R . P a g e , D.R. A b r a m o v i c h I, C . G . D a c k e 2, and A.Lee, Departments of Biomedical Sciences and Obstetrics and G y n a e c o l o g y I, A b e r d e e n U n i v e r s i t y , and S c h o o l of P h a r m a c y , P o r t s m o u t h U n i v e r s i t y 2. A l k a l i n e p h o s p h a t a s e (ALP) will c o m p e t e for ATP w i t h o t h e r enzymes, i n c l u d i n g ion pumps, in the p l a c e n t a l m i c r o v i l l o u s border. This s t u d y i n v e s t i g a t e s the e f f e c t s o f A L P i n h i b i t i o n on c a l c i u m (Ca) t r a n s p o r t into m i c r o v i l l o u s b o r d e r v e s i c l e s (MBV). 45Ca u p t a k e by MBV, i n c u b a t e d for p e r i o d s up to 30 m i n in i s o t o n i c s a l i n e c o n t a i n i n g I0 ~M Ca l a b e l l e d w i t h 45Ca was m e a s u r e d in the p r e s e n c e of the ALP i n h i b i t o r s l e v a m i s o l e (Lev) and p h e n y l a l a n i n e (Phe). Lev (i0 ~M) w h e n p r e s e n t i n s i d e the M B V p r o d u c e d a s i g n i f i c a n t 130% rise in 45Ca u p t a k e at 30 m i n ( p < 0 . 0 5 - p a i r e d t test, N=4). Lev (I0 pM) and A T P (0.5 mM) t o g e t h e r inside the M B V p r o d u c e d an i n c r e a s e in u p t a k e of 58% at 30 min. w h e n c o m p a r e d w i t h ATP (0.5 mM) alone. ATP (0.5 mM) inside p r o d u c e d no s i g n i f i c a n t c h a n g e in Ca uptake. U p t a k e at 30 min t e n d e d to i n c r e a s e w h e n e i t h e r L e v or Phe w e r e p r e s e n t at equal c o n c e n t r a t i o n on b o t h sides of the M B V m e m b r a n e s but s h o w e d no dose d e p e n d e n c e o v e r the r a n g e l0 ~M to 1 mM. O n l y the Phe i n d u c e d i n c r e a s e s w e r e s i g n i f i c a n t (p<0.05, N=I0). As the M B V w e r e o r i e n t a t e d w i t h the e x t r a c e l l u l a r s u r f a c e outward, these, r e s u l t s do not i n d i c a t e a m a j o r role in Ca t r a n s p o r t for an o u t w a r d l y directed Ca p u m p l o c a t e d on the p l a c e n t a l microvillous border.
CYTOKINES IN THE REPRODUCTION OF LOWER VERTEBRATES. aL. Paulesu, aR. Romagnoli, aM. Marchetti, bM. Cintorino, Cp. Ghiara & dG. Ghiara. aInstitute of General Physiology, bInstitute of Anatomy Pathology, University of Siena, CISIR, Siena, dDepartment of Evolutive Biology, University of Siena, Italy. A broad range of immunoregulatory peptides (cytokines) has been identified at the materno-fetal interface of different species of mammals either with invaeive or noninvasive type of placenta. Since some viviparous species of squamates (reptiles) have developed a non-invasive epitheliochorial placenta, we have examined whether cytokines, considered critical in mammalian pregnancy, could be also involved in the reproduction of lower vertebrates. Chorio-allantoic placentas of Cha~cides chalcides were incubated at 37~ in Dulbecco's Modified Eagle's Medium in an air/CO 2 atmosphere and the supernatants tested for the presence of interleukin 1 (IL-I) and transforming growth factor 81 (TGF-BI). Placental tissues were investigated by immunohistochemical analysis using alkaline phosphatase, anti-alkaline phosphatase (APAAP) technique and monoclonal antibodies anti-human IL-I u, IL-I B or TGF BI. The results have shown that IL-I and TGF B1 were released in the medium though at different amounts during incubation. Moreover immunoreactive cells for IL-I u, IL-I B and TGF B1 were observed in the chorial and the uterine epithelia with a different distribution throughout gestation. In the light of the data obtained in mammals, our data suggest that the presence of the cytokines at the maternofetal interface and their role in the maintenance of pregnancy should represent a critical event in the evolution of viviparity.
Placenta (1993), I/ol 14
E F F E C T S OF A L K A L I N E P H O S P H A T A S E I N H I B I T O R S U P O N R A D I O C A L C I U M UPTAKE BY M I C R O V I L L O U S BORDER MEMBRANES OF THE HUMAN P L A C E N T A . K . R . P a g e , D.R. A b r a m o v i c h I, C . G . D a c k e 2, and A.Lee, Departments of Biomedical Sciences and Obstetrics and G y n a e c o l o g y I, A b e r d e e n U n i v e r s i t y , and S c h o o l of P h a r m a c y , P o r t s m o u t h U n i v e r s i t y 2. A l k a l i n e p h o s p h a t a s e (ALP) will c o m p e t e for ATP w i t h o t h e r enzymes, i n c l u d i n g ion pumps, in the p l a c e n t a l m i c r o v i l l o u s border. This s t u d y i n v e s t i g a t e s the e f f e c t s o f A L P i n h i b i t i o n on c a l c i u m (Ca) t r a n s p o r t into m i c r o v i l l o u s b o r d e r v e s i c l e s (MBV). 45Ca u p t a k e by MBV, i n c u b a t e d for p e r i o d s up to 30 m i n in i s o t o n i c s a l i n e c o n t a i n i n g I0 ~M Ca l a b e l l e d w i t h 45Ca was m e a s u r e d in the p r e s e n c e of the ALP i n h i b i t o r s l e v a m i s o l e (Lev) and p h e n y l a l a n i n e (Phe). Lev (i0 ~M) w h e n p r e s e n t i n s i d e the M B V p r o d u c e d a s i g n i f i c a n t 130% rise in 45Ca u p t a k e at 30 m i n ( p < 0 . 0 5 - p a i r e d t test, N=4). Lev (I0 pM) and A T P (0.5 mM) t o g e t h e r inside the M B V p r o d u c e d an i n c r e a s e in u p t a k e of 58% at 30 min. w h e n c o m p a r e d w i t h ATP (0.5 mM) alone. ATP (0.5 mM) inside p r o d u c e d no s i g n i f i c a n t c h a n g e in Ca uptake. U p t a k e at 30 min t e n d e d to i n c r e a s e w h e n e i t h e r L e v or Phe w e r e p r e s e n t at equal c o n c e n t r a t i o n on b o t h sides of the M B V m e m b r a n e s but s h o w e d no dose d e p e n d e n c e o v e r the r a n g e l0 ~M to 1 mM. O n l y the Phe i n d u c e d i n c r e a s e s w e r e s i g n i f i c a n t (p<0.05, N=I0). As the M B V w e r e o r i e n t a t e d w i t h the e x t r a c e l l u l a r s u r f a c e outward, these, r e s u l t s do not i n d i c a t e a m a j o r role in Ca t r a n s p o r t for an o u t w a r d l y directed Ca p u m p l o c a t e d on the p l a c e n t a l microvillous border.
CYTOKINES IN THE REPRODUCTION OF LOWER VERTEBRATES. aL. Paulesu, aR. Romagnoli, aM. Marchetti, bM. Cintorino, Cp. Ghiara & dG. Ghiara. aInstitute of General Physiology, bInstitute of Anatomy Pathology, University of Siena, CISIR, Siena, dDepartment of Evolutive Biology, University of Siena, Italy. A broad range of immunoregulatory peptides (cytokines) has been identified at the materno-fetal interface of different species of mammals either with invaeive or noninvasive type of placenta. Since some viviparous species of squamates (reptiles) have developed a non-invasive epitheliochorial placenta, we have examined whether cytokines, considered critical in mammalian pregnancy, could be also involved in the reproduction of lower vertebrates. Chorio-allantoic placentas of Cha~cides chalcides were incubated at 37~ in Dulbecco's Modified Eagle's Medium in an air/CO 2 atmosphere and the supernatants tested for the presence of interleukin 1 (IL-I) and transforming growth factor 81 (TGF-BI). Placental tissues were investigated by immunohistochemical analysis using alkaline phosphatase, anti-alkaline phosphatase (APAAP) technique and monoclonal antibodies anti-human IL-I u, IL-I B or TGF BI. The results have shown that IL-I and TGF B1 were released in the medium though at different amounts during incubation. Moreover immunoreactive cells for IL-I u, IL-I B and TGF B1 were observed in the chorial and the uterine epithelia with a different distribution throughout gestation. In the light of the data obtained in mammals, our data suggest that the presence of the cytokines at the maternofetal interface and their role in the maintenance of pregnancy should represent a critical event in the evolution of viviparity.