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Heat shock proteins in mouse oogenesis
488
489
3H-thymidine Autoradiography of the duration of Meiosis and Spermiogenesis of b u l l , Bos indicus
FERRO-MAGNETIC ISOLATION OF MEMBRANES INVOLVED IN VITELLOGENIN TRANSPORT INTO XENOPUS OOCYTES A. Bauer and H.-P. R i c h t e r , FR 3.2 P h y s i o l o g i e , D-6650 Homburg/S. F.R.G. The pathway of the yolk precursor v i t e l l o g e n i n (VTG) has been followed by f e r r o - l a b e l l i n g , magnetic s o r t i n g , and EM. F e r r i c p a r t i c l e s of 10 nm are s e l e c t i v e l y t r a n s p o r t e d together with VTG i n t o the yolk p l a t e l e t s of Xenopus oocytes. Thus, several membrane compartments are involved in the i n c o r p o r a t i o n and subsequent t r a n s f e r of VTG. in i n t a c t oocytes, u n l a b e l l e d or l a b e l l e d with ferric particles, coated p i t s fuse with coated vesicl e s , coated v e s i c l e s fuse among themselves or with the c o r t i c a l ER. The r e s u l t i n g compartments discharge i n t o yolk endosomes. These endosomes are the direct precursors of the yolk o r g a n e l l e s . A f t e r pulse-chase loading with f e r r i c p a r t i c l e s and subsequent f r a c t i o n a t i o n of the oocytes, ferromagnetic sorting of the different v e s i c l e populations has been p e r f o r med using a " f r e e - f l o w magnetic chamb e r " . This novel method enables specification and c h a r a c t e r i z a t i o n of p u r i f i e d endosomal compartments t h a t accumulate r e c e p t o r - s p e c i f i c p r o t e i n .
T.K.BANERJEE, Zoology Dept.S.B.C~llege Magra Hooghly,W.B. India and S.K.GHOSAL,ZooIogy Dept.Burdwar, U n i v e r s i t y , W.B. India The course of p r o g r e s s i o n of 3H-thymidine labeled bovine spermatocytes was monitored a u t o r a d i o g r a p h i : c a l l y in vivo by applying Kodak AR-10 s t r i p p i n g film to both h i s t o l o g i c a l and a i r - d r i e d p r e p a r a t i o n s . An analy. s i s of data suggests that the duration of l e p t o t e n e and pachytene is about 3.8 and 16.8 days r e s p e e t i v e l y . l n sharp c o n t r a s t zygotene is of s h o r t e r duration and l a s t s for only 0.7 day.The t r a n s i t i o n of a l l stages from d i p l o tene to metaphase-ll is exceedingly f a s t and r e q u i r e s hardly one day.The duration of m e i o s i s , t h e r e f o r e , is about 1 6 . 3 days. Spermiogenesis, however takes a prolonged period and is completed within 20.5 days. The t o t a l duration of both meiosis and s p e r m i o g e n e s i s i s a b o u t 3 6 . 8 d a y s in Bos i n d i c u s .
490 HEAT S H O C K P R O T E I N S
491 Biochemical Differentiation in Mouse Testis During Postnatal Development. A.L. Bhatia and Sangeeta Pareek Present study provides some specific informa5ions on quantitative changes in chemical constituents of the mouse testis during maturation. Postnatally developing mice testes were studied for biochemical differentiation. Litters were autopsied at i-6 weeks p.p., their testes were taken out and studied for cholesterol, DNA, protein and various enzymes. The ~estis and body weight both exhibited a sharp increase from 3 weeks onward. The testis growth rate exceeded chat of body weight at this interval, ire total protein content remained relatively constant at 1-3 weeks of age followed by a sharp rise at 4 weeks, which again declined by 5th week and maintained a steady state onward. DNA concentration occurred maximum at 1 week of age and declined cill 6th week. An elevated cholesterol concentration at 1 week of age sharply declined oy 4th week followed by a gradual rise to the adult value. A gradual decrease in acid phosphatase activity from 1 week ~o 4 weeks of age has been noticed which increased e little and attained a steady state on the later interval Alkaline p~osphatase too declined gradually till 3 weeks of age, being sharp onward. Adenosine triphosphatase activity showed a oeak on 3rd week followed by a dip at 4 weeks of age.
IN MOIIBE O O G E N E S ! S
Curc~, A., Bevilacqua. A.. and Manqia. F. Institute of Histology and general Embryology and Dept. of Psychology, University "La Sapienza" of Rome, Italy.
It was previously demonstrated in our laboratory that preovulatory mammalian oocytes constitutively synthesize heat shock protein B9 (HSP89) and heat shock cognate protein 70 (HSC70), but lack an inducible HSP synthesis in response to hyperthermia (Curci et al., Dev. Biol. 123, !54-160, 1987). We have now investigated the pattern of H~-gene expression in growing mouse oocytes. Similarly to the preovulatory cells, growing oocytes constitutively synthesized HSP89 and HSC70, and} when heat shocked, displayed induction of isolelectric variants of HSP68. HSP68-inducibility was dev~l~pmentally regulated during oocyte growth and was experimentally suppressed by an in vivo administration of PMSG. We propose that HS-response in mam~nalian female germ cells is suppressed at the end of oogenesis by the gonadotropin-dependent development of the Graafian follicle.
S150
489
3H-thymidine Autoradiography of the duration of Meiosis and Spermiogenesis of b u l l , Bos indicus
FERRO-MAGNETIC ISOLATION OF MEMBRANES INVOLVED IN VITELLOGENIN TRANSPORT INTO XENOPUS OOCYTES A. Bauer and H.-P. R i c h t e r , FR 3.2 P h y s i o l o g i e , D-6650 Homburg/S. F.R.G. The pathway of the yolk precursor v i t e l l o g e n i n (VTG) has been followed by f e r r o - l a b e l l i n g , magnetic s o r t i n g , and EM. F e r r i c p a r t i c l e s of 10 nm are s e l e c t i v e l y t r a n s p o r t e d together with VTG i n t o the yolk p l a t e l e t s of Xenopus oocytes. Thus, several membrane compartments are involved in the i n c o r p o r a t i o n and subsequent t r a n s f e r of VTG. in i n t a c t oocytes, u n l a b e l l e d or l a b e l l e d with ferric particles, coated p i t s fuse with coated vesicl e s , coated v e s i c l e s fuse among themselves or with the c o r t i c a l ER. The r e s u l t i n g compartments discharge i n t o yolk endosomes. These endosomes are the direct precursors of the yolk o r g a n e l l e s . A f t e r pulse-chase loading with f e r r i c p a r t i c l e s and subsequent f r a c t i o n a t i o n of the oocytes, ferromagnetic sorting of the different v e s i c l e populations has been p e r f o r med using a " f r e e - f l o w magnetic chamb e r " . This novel method enables specification and c h a r a c t e r i z a t i o n of p u r i f i e d endosomal compartments t h a t accumulate r e c e p t o r - s p e c i f i c p r o t e i n .
T.K.BANERJEE, Zoology Dept.S.B.C~llege Magra Hooghly,W.B. India and S.K.GHOSAL,ZooIogy Dept.Burdwar, U n i v e r s i t y , W.B. India The course of p r o g r e s s i o n of 3H-thymidine labeled bovine spermatocytes was monitored a u t o r a d i o g r a p h i : c a l l y in vivo by applying Kodak AR-10 s t r i p p i n g film to both h i s t o l o g i c a l and a i r - d r i e d p r e p a r a t i o n s . An analy. s i s of data suggests that the duration of l e p t o t e n e and pachytene is about 3.8 and 16.8 days r e s p e e t i v e l y . l n sharp c o n t r a s t zygotene is of s h o r t e r duration and l a s t s for only 0.7 day.The t r a n s i t i o n of a l l stages from d i p l o tene to metaphase-ll is exceedingly f a s t and r e q u i r e s hardly one day.The duration of m e i o s i s , t h e r e f o r e , is about 1 6 . 3 days. Spermiogenesis, however takes a prolonged period and is completed within 20.5 days. The t o t a l duration of both meiosis and s p e r m i o g e n e s i s i s a b o u t 3 6 . 8 d a y s in Bos i n d i c u s .
490 HEAT S H O C K P R O T E I N S
491 Biochemical Differentiation in Mouse Testis During Postnatal Development. A.L. Bhatia and Sangeeta Pareek Present study provides some specific informa5ions on quantitative changes in chemical constituents of the mouse testis during maturation. Postnatally developing mice testes were studied for biochemical differentiation. Litters were autopsied at i-6 weeks p.p., their testes were taken out and studied for cholesterol, DNA, protein and various enzymes. The ~estis and body weight both exhibited a sharp increase from 3 weeks onward. The testis growth rate exceeded chat of body weight at this interval, ire total protein content remained relatively constant at 1-3 weeks of age followed by a sharp rise at 4 weeks, which again declined by 5th week and maintained a steady state onward. DNA concentration occurred maximum at 1 week of age and declined cill 6th week. An elevated cholesterol concentration at 1 week of age sharply declined oy 4th week followed by a gradual rise to the adult value. A gradual decrease in acid phosphatase activity from 1 week ~o 4 weeks of age has been noticed which increased e little and attained a steady state on the later interval Alkaline p~osphatase too declined gradually till 3 weeks of age, being sharp onward. Adenosine triphosphatase activity showed a oeak on 3rd week followed by a dip at 4 weeks of age.
IN MOIIBE O O G E N E S ! S
Curc~, A., Bevilacqua. A.. and Manqia. F. Institute of Histology and general Embryology and Dept. of Psychology, University "La Sapienza" of Rome, Italy.
It was previously demonstrated in our laboratory that preovulatory mammalian oocytes constitutively synthesize heat shock protein B9 (HSP89) and heat shock cognate protein 70 (HSC70), but lack an inducible HSP synthesis in response to hyperthermia (Curci et al., Dev. Biol. 123, !54-160, 1987). We have now investigated the pattern of H~-gene expression in growing mouse oocytes. Similarly to the preovulatory cells, growing oocytes constitutively synthesized HSP89 and HSC70, and} when heat shocked, displayed induction of isolelectric variants of HSP68. HSP68-inducibility was dev~l~pmentally regulated during oocyte growth and was experimentally suppressed by an in vivo administration of PMSG. We propose that HS-response in mam~nalian female germ cells is suppressed at the end of oogenesis by the gonadotropin-dependent development of the Graafian follicle.
S150