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稀释培养基作为淡水虹鳟鱼培养鳃上皮细胞中的环境或生理模拟物。

Dilute culture media as an environmental or physiological simulant in cultured gill epithelia from freshwater rainbow trout.

作者信息

Kelly Scott P, Wood Chris M

机构信息

Department of Biology, McMaster University, Hamilton, Ontario, Canada L8S 1S1.

出版信息

In Vitro Cell Dev Biol Anim. 2003 Jan-Feb;39(1-2):21-8. doi: 10.1290/1543-706X(2003)039<0021:DCMAAE>2.0.CO;2.

Abstract

The electrophysiological and ion-transporting properties of cultured gill epithelia from freshwater (FW) rainbow trout were examined in the presence of dilute cell culture media as an environmental or physiological simulant. Gill epithelia were cultured on cell culture inserts under symmetrical conditions (L15 apical-L15 basolateral) for 6-7 d. The following experiments were then conducted. (1) To mimic a gradual lowering of environmental salinity, apical L15 medium was progressively diluted with FW (first to 2/3 L15 for 8 h and then to 1/3 L15 for 6 h) before the introduction of apical FW (FW apical-L15 basolateral, analogous to a fish in a natural FW environment). Dilute apical media had no significant effect on the electrophysiological properties of preparations compared with symmetrical culture conditions, and no evidence for active Na(+) or Cl(-) transport was observed. Preparations subsequently exposed to apical FW exhibited a negative transepithelial potential and evidence of active Cl(-) uptake and slight Na(+) extrusion. (2) To mimic the extracellular fluid dilution that occurs in euryhaline fish after abrupt transfer from saline to FW, the osmolality or ionic strength (or both) of basolateral media was reduced by 20-40% (using either FW or FW + mannitol) while simultaneously replacing apical media with FW. Under these conditions, Na(+) and Cl(-) influx rates were low compared with efflux rates, while the Ussing flux ratio analysis generally indicated active Cl(-) uptake and Na(+) extrusion. The Na(+)-K(+) adenosine triphosphatase activity was not affected by alterations in basolateral osmolality. Our studies indicate that cultured trout gill epithelia are tolerant of media dilution from both the apical and the basolateral direction; however, neither treatment alone appeared to increase ion influx rates or stimulate active Na(+) uptake in cultured trout gill epithelia.

摘要

在稀释的细胞培养基作为环境或生理模拟物存在的情况下,研究了淡水(FW)虹鳟鱼培养鳃上皮的电生理和离子转运特性。鳃上皮在细胞培养插入物上于对称条件下(L15顶侧 - L15基底侧)培养6 - 7天。然后进行了以下实验。(1)为模拟环境盐度的逐渐降低,在引入顶侧淡水(FW顶侧 - L15基底侧,类似于处于天然淡水环境中的鱼)之前,先用淡水将顶侧L15培养基逐步稀释(先稀释至2/3 L15持续8小时,然后稀释至1/3 L15持续6小时)。与对称培养条件相比,稀释的顶侧培养基对制剂的电生理特性没有显著影响,并且未观察到主动Na(+)或Cl(-)转运的证据。随后暴露于顶侧淡水的制剂表现出负的跨上皮电位以及主动Cl(-)摄取和轻微Na(+)排出的证据。(2)为模拟广盐性鱼类从盐水突然转移到淡水后发生的细胞外液稀释,通过用淡水或淡水 + 甘露醇将基底侧培养基的渗透压或离子强度(或两者)降低20 - 40%,同时用淡水替换顶侧培养基。在这些条件下,与流出速率相比,Na(+)和Cl(-)流入速率较低,而尤斯辛通量比率分析通常表明有主动Cl(-)摄取和Na(+)排出。Na(+)-K(+)三磷酸腺苷酶活性不受基底侧渗透压变化的影响。我们的研究表明,培养的鳟鱼鳃上皮对来自顶侧和基底侧方向的培养基稀释具有耐受性;然而,单独的任何一种处理似乎都不会增加培养的鳟鱼鳃上皮中的离子流入速率或刺激主动Na(+)摄取。

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