Anti-proteoglycan antibodies that give perineuronal staining of a subset of central neurons

Anti-proteoglycan antibodies that give perineuronal staining of a subset of central neurons

S114 ANTI-PROTEOGLYCAN ANTIBODIES THAT GIVE PERINEURONAL STAINING OF A SUBSET OF CENTRAL NEURONS. SHINOBU C. FUJITA I Department of PharmacoloKy, Gunm...

55KB Sizes 1 Downloads 33 Views

S114 ANTI-PROTEOGLYCAN ANTIBODIES THAT GIVE PERINEURONAL STAINING OF A SUBSET OF CENTRAL NEURONS. SHINOBU C. FUJITA I Department of PharmacoloKy, Gunma Universit2 School of Medicine t Maebashi 371~ Japan. Previously we described three monoclonal antibodies (MAbs 473, 376 and 528) obtained by immunization with extracts of monkey brain. These three MAbs give perineuronal staining of subsets of mammalian central neurons, and their immunoreactivities are lost by pretreatment of the sections with chondroitinase ABC. We have now tested 13 MAbs with claimed specificity to proteoglycans; three give similar perineuronal staining of subsets of central neurons in the monkey, rat and mouse. These are the MAbs IB5, 9A2 and 3B3 of Couchman et al. (1984) with established specificity to chondroitinase ABC-revealed epitopes associated with sulfated glycosaminoglycans. Dual color i n d i r e c t immunofluorescence staining showed that neurons positive to MAbs 9A2 or 3B3 are generally also positive to MAb IB5. Neurons positive to MAbs 473 or 376 were also positive to MAb IB5. Occurrence of chondroitinase-inactivated epitopes of MAbs 473 and 376 on neurons with chondreitinase-revealed glycosaminoglycan epitope of MAb 1B5 suggests that the antigens bearing the former epitopes are proteoglycans themselves or some closely related substance.

PHOSPHORUS POTASSIUM Department

NUCLEAR M A G N E T I C R E S O N A N C E , S T U D I E S ON R A T B R A I N SLICES: EFFECT OF DEPOLARIZATION. M I T S U O TAKEI, Y O S H I H I S A KAWANO* AND KAZUHIRO YAMADA* o f P h y s i o l o g y , M e d i c a l C o l l e g e o f oita, o i t a 879-56, Japan

we p e r f o r m e d p h o s p h o r u s n u c l e a r m a g n e t i c r e s o n a n c e (31p-NMR) s t u d i e s on rat brain s l i c e s (dry w e i g h t 0.4g) s u p e r f u s e d w i t h n o r m a l and h i g h p o t a s s i u m Krebs solutions. S p e c t r a were o b h a i n e d by a c c u m u l a t i n g 64 scans w i t h an interpulse i n t e r v a l of 4 s (time r e s o l u t i o n 4 min). After depolarization i n d u c e d by 60 mM potassium, the peak amplitude of p h o s p h o c r e ~ t i n e (PCr) decreased, that of inorganic p h o s p h a t e (P.) i ~ c r e a s e d a n d an a c i d i e shift of i n t r a c e l l u l a r pH was 1 observed. These changes r e t u r n e d to i n i t i a l l e v e l s w i t h i n i0 or 15 mln in normal Krebs solution, and this e f f e c t of h i g h p o t a s s i u m could be observed repeatedly on the same t i s s u e p r e p a r a t i o n . S i m i l a r but lesser effects were observed w i t h 30 and 40 m M p o t a s s i u m . W h e n o u a b a i n (0.i-i.0 mM) or T T X (i0 g ml- ) was p r e s e n t in the s u p e r f u s i n g media, h i g h p o t a s s i u m p r o d u c e d the same e f f e c t as seen w i t h o u t t h e s e drugs. T h e s e r e s u l t s i n d i c a t e that e n e r g y f r o m PCr s p l i t t i n g is not c o n s u m e d by the N a - K pump. m-conotoxin, a Ca c h a n n e l (N type) blocker, also had no e f f e c t on PCr s p l i t t i n g .