Identification of Sodium- and Chloride-Sensitive Sites in the Slack Channel.

The Slack channel (KCNT1, Slo2.2) is a sodium-activated and chloride-activated potassium channel that regulates heart rate and maintains the normal excitability of the nervous system. Despite intense interest in the sodium gating mechanism, a comprehensive investigation to identify the sodium-sensitive and chloride-sensitive sites has been missing. In the present study, we identified two potential sodium-binding sites in the C-terminal domain of the rat Slack channel by conducting electrophysical recordings and systematic mutagenesis of cytosolic acidic residues in the rat Slack channel C terminus. In particular, by taking advantage of the M335A mutant, which results in the opening of the Slack channel in the absence of cytosolic sodium, we found that among the 92 screened negatively charged amino acids, E373 mutants could completely remove sodium sensitivity of the Slack channel. In contrast, several other mutants showed dramatic decreases in sodium sensitivity but did not abolish it altogether. Furthermore, molecular dynamics (MD) simulations performed at the hundreds of nanoseconds timescale revealed one or two sodium ions at the E373 position or an acidic pocket composed of several negatively charged residues. Moreover, the MD simulations predicted possible chloride interaction sites. By screening predicted positively charged residues, we identified R379 as a chloride interaction site. Thus, we conclude that the E373 site and the D863/E865 pocket are two potential sodium-sensitive sites, while R379 is a chloride interaction site in the Slack channel.SIGNIFICANCE STATEMENT The research presented here identified two distinct sodium and one chloride interaction sites located in the intracellular C-terminal domain of the Slack (Slo2.2, KCNT1) channel. Identification of the sites responsible for the sodium and chloride activation of the Slack channel sets its gating property apart from other potassium channels in the BK channel family. This finding sets the stage for future functional and pharmacological studies of this channel.

Characteristics of phytoplankton community and its influencing factors in the Yanhe River Basin, Northwest China.

Yanhe River is one of the important tributaries of the Yellow River, with a vital role in the maintenance of biodiversity and ecological conservation in the middle reaches of the Yellow River. In this study, we conducted a systematic aquatic ecological survey of the Yanhe River Basin in spring (April-May) and autumn (September-October) of 2021, with phytoplankton as indicator organism. A total of 33 sampling sections were selected in the mainstem, five first-class tributaries, and impounded water bodies (reservoir and check dam water bodies) of the Yanhe River Basin. The results showed that a total of 253 phytoplankton species, belonging to 7 phyla and 91 genera, were detected in the two surveys. Diatoms and green algae prevailed in spring (168 species), while diatoms and cyanobacteria dominated in autumn (179 species). The mean phytoplankton density and biomass were 316.07×104 cells·L-1 and 6.41 mg·L-1 in spring, and 69.56×104 cells·L-1 and 1.59 mg·L-1 in autumn, respectively. At the temporal scale, phytoplankton abundance in spring was higher than that in autumn. At the spatial scale, the phytoplankton abundance in the middle and lower reaches of the mainstream was higher than that in the upper reaches. Phytoplankton biomass in the impounded water bodies formed by dam interception was maintained at a high level, which was significantly higher than that in the mainstem and tributary water bodies in autumn. The phytoplankton diversity, as indicated by Shannon diversity index, Margalef richness index, and Pielou evenness index, in spring was greater than that in autumn. Phytoplankton diversity was greater in the trunk and tributary waters than that in impounded waters. The results of redundancy analysis showed that the key factors driving the phytoplankton community structure in spring were flow velocity, dissolved oxygen, nitrite nitrogen, and water depth. In contrast, the key driving factors in autumn were nitrate nitrogen, water depth, and dissolved oxygen.