The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1 ~3in the Basi lar Membrane of Neonatal Rat Cochlea
10.3969/j.issn.1006-7299.2016.04.011
- VernacularTitle:质膜钙ATP酶异构体1～3在新生大鼠耳蜗基底膜的表达
- Author:
Qingguo CHEN
;
Hanqi CHU
;
Liangqiang ZHOU
;
Jin CHEN
;
Yun LIU
;
Mi LUO
;
Yanling TAO
- Publication Type:Journal Article
- Keywords:
Plasma membrane Ca2+-ATPase isoform 1 (PMCA1 );
Plasma membrane Ca2+-ATPase iso-forms 2(PMCA2);
Plasma membrane Ca2+-ATPase isoforms 3(PMCA3);
Basilar membrane;
Cochlea
- From:
Journal of Audiology and Speech Pathology
2016;24(4):366-370
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the expression of plasma membrane Ca2+-ATPase isoforms 1~3 (PMCA 1~3 )in the basilar membrane (BM)of the neonatal rat cochlea by Western blot.The PMCA2 content in single BM of the neonatal rat was also examined.Methods Four rats at postnatal 2 days (P2)and 8 days (P8)were respective-ly decapitated and their BMs were isolated.The total proteins of BMs were extracted.The 20μg total proteins were respectively loaded to the gel.The expression of PMCA1-3 was detected by Western blot.Likewise,3μg total proteins from P2 and P8 rat BM were loaded.The expression of PMCA2 was detected by Western blot.Four rats at P8 were decapitated and their BM was isolated.The 5μg,10μg and 20μg total proteins of P8 rat BM were added to the gel and 100 ng,400 ng and 800 ng bovine serum albumin (BSA)were also loaded as reference.After electro-phoresis,the gel was separated into two parts.One part was used for SYPRO staining and the other part was used for PMCA2 detection by Western blot.Results In the 20μg BM total proteins of P2 and P8 rats,the expression of PMCA1 was weak (0.126±0.024,0.131±0.012,respectively),PMCA2 was strong (4.16±0.528,4.25±0.319, respectively),and PMCA3 was barely expressed (0 ).There was a statistical difference among PMCA1 ,PMCA2 and PMCA3(P<0.05).In the 3μg BM total proteins of P2 and P8 rats,the expression of PMCA2 in P8 (4.571± 0.336)was higher than P2 (3.622±0.285).There was a statistical difference(P<0.05).The PMCA2 content in the BM of a P8 rat was about 2 .5 ng.Conclusion There was a different-level expression of PMCA1~3 in the neonatal rat BM with highest expression of PMCA2 ,which could be explained that cochlear hair cells had different requirements of Ca2+ turning for these PMCA isoforms.
