{"title":"大鼠腮腺基底侧质膜囊Ca2+通透性受膜电位和囊外[Ca2+]调节。","authors":"T Lockwich, I S Ambudkar, A E Shamoo","doi":"10.3109/09687689309150264","DOIUrl":null,"url":null,"abstract":"<p><p>This study examines the Ca2+ permeability of basolateral plasma membrane vesicles (BLMVs) isolated from the rat parotid gland by monitoring the rate of 45Ca2+ efflux from actively-loaded (via the Ca(2+)-ATPase) inside-out BLMVs. Ca2+ efflux from BLMVs into a K(+)-gluconate medium which hyperpolarizes the cytoplasmic side (i.e. outside) of the inside-out BLMVs resulted in a faster rate of Ca2+ efflux compared with a control medium containing N-methyl-D-glucamine (NMDG)-gluconate. Conversely, Ca2+ efflux into a medium which depolarizes the cytoplasmic side of the BLMVs (NMDG-chloride) resulted in slower rates of efflux compared with those observed with the control medium. This increased rate of 45Ca2+ efflux from the hyperpolarized BLMV was inhibited by 1 mM Ni2+, yielding a rate of efflux similar to the rate observed in depolarized BLMVs. The rate of Ca2+ efflux from BLMVs was affected by [Ca2+]o ([Ca2+] on the extravesicular, cytoplasmic side of the vesicle). When [Ca2+]o was kept > 200 nM during efflux, the rate of Ca2+ efflux from both hyper- and depolarized BLMVs was slow and relatively unresponsive to changes in [Ca2+]o, despite sizeable changes in the Ca2+ gradient across the BLMV. However, when [Ca2+]o was lowered < 200 nM, there was an abrupt increase in the rate of Ca2+ efflux from both hyper- and depolarized BLMVs. Additionally, when [Ca2+] was < 200 nM, the rate of Ca2+ efflux appeared to be more sensitive to driving force changes. These data suggest that Ca2+ permeability across the rat parotid gland basolateral plasma membrane is modulated by membrane potential and [Ca2+] on the cytoplasmic side.</p>","PeriodicalId":18448,"journal":{"name":"Membrane biochemistry","volume":"10 3","pages":"171-9"},"PeriodicalIF":0.0000,"publicationDate":"1993-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/09687689309150264","citationCount":"3","resultStr":"{\"title\":\"Ca2+ permeability of rat parotid gland basolateral plasma membrane vesicles is modulated by membrane potential and extravesicular [Ca2+].\",\"authors\":\"T Lockwich, I S Ambudkar, A E Shamoo\",\"doi\":\"10.3109/09687689309150264\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>This study examines the Ca2+ permeability of basolateral plasma membrane vesicles (BLMVs) isolated from the rat parotid gland by monitoring the rate of 45Ca2+ efflux from actively-loaded (via the Ca(2+)-ATPase) inside-out BLMVs. Ca2+ efflux from BLMVs into a K(+)-gluconate medium which hyperpolarizes the cytoplasmic side (i.e. outside) of the inside-out BLMVs resulted in a faster rate of Ca2+ efflux compared with a control medium containing N-methyl-D-glucamine (NMDG)-gluconate. Conversely, Ca2+ efflux into a medium which depolarizes the cytoplasmic side of the BLMVs (NMDG-chloride) resulted in slower rates of efflux compared with those observed with the control medium. This increased rate of 45Ca2+ efflux from the hyperpolarized BLMV was inhibited by 1 mM Ni2+, yielding a rate of efflux similar to the rate observed in depolarized BLMVs. The rate of Ca2+ efflux from BLMVs was affected by [Ca2+]o ([Ca2+] on the extravesicular, cytoplasmic side of the vesicle). When [Ca2+]o was kept > 200 nM during efflux, the rate of Ca2+ efflux from both hyper- and depolarized BLMVs was slow and relatively unresponsive to changes in [Ca2+]o, despite sizeable changes in the Ca2+ gradient across the BLMV. However, when [Ca2+]o was lowered < 200 nM, there was an abrupt increase in the rate of Ca2+ efflux from both hyper- and depolarized BLMVs. Additionally, when [Ca2+] was < 200 nM, the rate of Ca2+ efflux appeared to be more sensitive to driving force changes. These data suggest that Ca2+ permeability across the rat parotid gland basolateral plasma membrane is modulated by membrane potential and [Ca2+] on the cytoplasmic side.</p>\",\"PeriodicalId\":18448,\"journal\":{\"name\":\"Membrane biochemistry\",\"volume\":\"10 3\",\"pages\":\"171-9\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1993-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.3109/09687689309150264\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Membrane biochemistry\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.3109/09687689309150264\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Membrane biochemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3109/09687689309150264","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3
摘要
本研究检测了从大鼠腮腺分离的基底外侧质膜囊(blmv)的Ca2+通透性,通过监测45Ca2+外排率(通过Ca(2+)- atp酶)内外blmv。与含有n -甲基- d -葡萄糖胺(NMDG)-葡萄糖酸盐的对照培养基相比,钙离子从blmv外排到K(+)-葡萄糖酸盐培养基中,该培养基使blmv的细胞质侧(即外部)超极化,导致Ca2+外排速度更快。相反,Ca2+外排到blmv细胞质侧去极化的介质(NMDG-chloride)中,与在对照介质中观察到的情况相比,导致外排速率较慢。从超极化BLMV中增加的45Ca2+外排速率被1 mM Ni2+抑制,产生的外排速率与去极化BLMV中观察到的速率相似。blmv的Ca2+外排速率受到[Ca2+]o ([Ca2+]在囊泡外的细胞质侧)的影响。当[Ca2+]o在外排过程中保持> 200 nM时,超极化和去极化BLMV的Ca2+外排速率缓慢且相对不响应[Ca2+]o的变化,尽管整个BLMV的Ca2+梯度变化很大。然而,当[Ca2+]o降低到< 200 nM时,超极化和去极化blmv的Ca2+外排速率都突然增加。此外,当[Ca2+] < 200 nM时,Ca2+外排速率对驱动力变化更为敏感。这些数据表明,通过大鼠腮腺基底侧质膜Ca2+通透性是由膜电位和细胞质侧的[Ca2+]调节的。
Ca2+ permeability of rat parotid gland basolateral plasma membrane vesicles is modulated by membrane potential and extravesicular [Ca2+].
This study examines the Ca2+ permeability of basolateral plasma membrane vesicles (BLMVs) isolated from the rat parotid gland by monitoring the rate of 45Ca2+ efflux from actively-loaded (via the Ca(2+)-ATPase) inside-out BLMVs. Ca2+ efflux from BLMVs into a K(+)-gluconate medium which hyperpolarizes the cytoplasmic side (i.e. outside) of the inside-out BLMVs resulted in a faster rate of Ca2+ efflux compared with a control medium containing N-methyl-D-glucamine (NMDG)-gluconate. Conversely, Ca2+ efflux into a medium which depolarizes the cytoplasmic side of the BLMVs (NMDG-chloride) resulted in slower rates of efflux compared with those observed with the control medium. This increased rate of 45Ca2+ efflux from the hyperpolarized BLMV was inhibited by 1 mM Ni2+, yielding a rate of efflux similar to the rate observed in depolarized BLMVs. The rate of Ca2+ efflux from BLMVs was affected by [Ca2+]o ([Ca2+] on the extravesicular, cytoplasmic side of the vesicle). When [Ca2+]o was kept > 200 nM during efflux, the rate of Ca2+ efflux from both hyper- and depolarized BLMVs was slow and relatively unresponsive to changes in [Ca2+]o, despite sizeable changes in the Ca2+ gradient across the BLMV. However, when [Ca2+]o was lowered < 200 nM, there was an abrupt increase in the rate of Ca2+ efflux from both hyper- and depolarized BLMVs. Additionally, when [Ca2+] was < 200 nM, the rate of Ca2+ efflux appeared to be more sensitive to driving force changes. These data suggest that Ca2+ permeability across the rat parotid gland basolateral plasma membrane is modulated by membrane potential and [Ca2+] on the cytoplasmic side.