Stevanović Katarina S, Čepkenović Bogdana, Križak Strahinja, Živić Miroslav Ž, Todorović Nataša V
Faculty of Biology, Institute of Physiology and Biochemistry, University of Belgrade, Studentski Trg 16, 11158 Belgrade, Serbia.
Institute of Multidisciplinary Research, University of Belgrade, Kneza Višeslava 1, 11030 Belgrade, Serbia.
J Fungi (Basel). 2023 May 31;9(6):637. doi: 10.3390/jof9060637.
Studies of ion currents in filamentous fungi are a prerequisite for forming a complete understanding of their physiology. Cytoplasmic droplets (CDs), obtained from sporangiophores of Phycomyces blakesleeanus, are a model system that enables the characterization of ion currents in the native membrane, including the currents mediated by the channels not yet molecularly identified. Osmotically activated anionic current with outward rectification (ORIC) is a dominant current in the membrane of cytoplasmic droplets under the conditions of hypoosmotic stimulation. We have previously reported remarkable functional similarities of ORIC with the vertebrate volume regulated anionic current (VRAC), such as dose-dependent activation by osmotic difference, ion selectivity sequence, and time and voltage dependent profile of the current. Using the patch clamp method on the CD membrane, we further resolve VRAC-like ORIC characteristics in this paper. We examine the inhibition by extracellular ATP and carbenoxolone, the permeation of glutamate in presence of chloride, selectivity for nitrates, and activation by GTP, and we show its single channel behavior in excised membrane. We propose that ORIC is a functional counterpart of vertebrate VRAC in filamentous fungi, possibly with a similar essential role in anion efflux during cell volume regulation.
对丝状真菌离子电流的研究是全面了解其生理学的先决条件。从布氏梨形孢的孢子梗获得的细胞质液滴(CDs)是一种模型系统,能够对天然膜中的离子电流进行表征,包括由尚未在分子水平上鉴定的通道介导的电流。在低渗刺激条件下,具有外向整流的渗透激活阴离子电流(ORIC)是细胞质液滴膜中的主要电流。我们之前报道过ORIC与脊椎动物容积调节性阴离子电流(VRAC)具有显著的功能相似性,例如由渗透压差引起的剂量依赖性激活、离子选择性序列以及电流的时间和电压依赖性特征。在本文中,我们使用膜片钳技术对CD膜进行研究,进一步解析了类似VRAC的ORIC特性。我们研究了细胞外ATP和甘草次酸的抑制作用、氯离子存在时谷氨酸的通透情况、对硝酸盐的选择性以及GTP的激活作用,并展示了其在分离膜中的单通道行为。我们提出,ORIC是丝状真菌中脊椎动物VRAC的功能对应物,可能在细胞容积调节过程中的阴离子外流中发挥类似的重要作用。