Center for Marine Research, Institute Ruđer Bošković, Rovinj, Zagreb, Croatia.
Alfred Wegener-Institute Helmholtz Centre for Polar and Marine Research, Wadden Sea Station List, Sylt, Germany.
J Comp Physiol B. 2020 Jul;190(4):391-401. doi: 10.1007/s00360-020-01277-2. Epub 2020 Apr 24.
The gills are osmorespiratory organs of aquatic organisms and the prime target of environmentally induced hypoxia. We have studied the impact of severe hypoxia (0.5 mg O/L) on the ionic transport across posterior gills of Carcinus maenas acclimated to 12 ppt seawater (DSW). The short-circuit current (Isc) across hemilamellae from gills i.e. active ion transport was studied in micro Ussing chambers. Hypoxia induced by deoxygenation of the basolateral side, and not the apical side, resulted in time-dependent inhibition of Isc and full recovery of Isc after reoxygenation. Exposure of the crabs to severe 7 h hypoxia decreased the specific activity of Na,K-ATPase in the gills by 36%. Full recovery of enzyme activity occurred in fasted crabs after 3 days of reoxygenation. The intensity of Western blotting bands was not different in the gills of oxygenated, hypoxic and reoxygenated crabs. The reversible, nonspecific blocker of K channels Cs and hypoxia inhibited over 90% of Isc which is after reoxygenation fully recovered. The specific blocker of Cl channels NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] blocked Isc by 68.5%. Only the rest of not inhibited Isc in aerated saline was blocked by hypoxia and recovered after reoxygenation. The activity of the antioxidant enzyme catalase was not changed during hypoxia and reoxygenation kept the high enzyme activity in the gills at the level of crabs acclimated to DSW. As a response to hypoxia the presence of 2 mM HO induce an initial slight transient decrease of Isc followed by a rise and after reoxygenation fully recovered Isc. Incubation of hemilamellae with the antioxidant derivative Trolox did not affect the inhibition of Isc by hypoxia.
鳃是水生生物的渗透呼吸器官,也是环境诱导缺氧的主要目标。我们研究了严重缺氧(0.5 mg O/L)对适应 12 ppt 海水(DSW)的 Carcinus maenas 后鳃离子转运的影响。通过微 Ussing 室研究了来自鳃的半鳃片的短路电流(Isc),即主动离子转运。通过基底外侧缺氧而不是顶端缺氧诱导的缺氧导致 Isc 呈时间依赖性抑制,并在再氧化后完全恢复 Isc。将螃蟹暴露于严重的 7 h 缺氧中会使鳃中的 Na,K-ATPase 比活力降低 36%。在再氧化 3 天后,禁食螃蟹的酶活性完全恢复。在充氧、缺氧和再氧化螃蟹的鳃中,Western blot 带的强度没有差异。K 通道的可逆、非特异性阻断剂 Cs 和缺氧抑制了超过 90%的 Isc,再氧化后完全恢复。Cl 通道的特异性阻断剂 NPPB [5-硝基-2-(3-苯基丙基氨基)苯甲酸] 阻断了 68.5%的 Isc。仅在充气盐水中未被抑制的 Isc 的其余部分被缺氧阻断,并在再氧化后恢复。抗氧化酶过氧化氢酶的活性在缺氧期间没有改变,并且再氧化使适应 DSW 的螃蟹的鳃中保持高酶活性。作为对缺氧的反应,存在 2 mM HO 会导致初始轻微的瞬态 Isc 下降,随后上升,再氧化后完全恢复 Isc。将半鳃片与抗氧化剂衍生物 Trolox 孵育不会影响缺氧对 Isc 的抑制作用。