Deficient LRRC8A-dependent volume-regulated anion channel activity is associated with male infertility in mice

Bao, Jianqiang; Pérez, Claudio F.; Kim, Jeesun; Zhang, Huan; Murphy, Caitlin J.; Hamidi, Tewfik; Jaubert, Jean; Platt, Craig D.; Chou, Janet; Deng, Mei; Zhou, Miou; Huang, Yuying; Gaitán‐Peñas, Héctor; Guénet, Jean Louis; Lin, Kevin; Lu, Yue; Chen, Taiping; Bedford, Mark T.; Dent, Sharon Y.R.; Richburg, John H.; Estévez, Raúl; Pan, Hui Lin; Geha, Raif S.; Shi, Qinghua; Benavides, Fernando

doi:10.1172/jci.insight.99767

Cited by 29 publications

(28 citation statements)

References 58 publications

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“…However, the genes encoding the channels remained elusive and highly controversial until 2014 when two independent groups definitively identified the Leucine‐Rich Repeat Containing 8A‐E (LRRC8A‐E) genes that encode VRACs (Kumar et al, ; Voss et al, ) Results from gene disruption, heterologous expression, and cryo‐EM experiments indicate that VRACs are likely hexameric channels that contain LRRC8A and at least one other LRRC8 subunit (i.e., LRRC8B, LRRC8C, LRRC8D, or LRRC8E) (Strange et al, ). Importantly for the present study, several groups have shown that genetic disruption of the LRRC8A gene abolishes VRAC activity in a variety of different cell types (Bao et al, ; Platt et al, ; Stuhlmann, Planells‐Cases, & Jentsch, ; Trothe et al, ).…”

Section: Introductionmentioning

confidence: 54%

The LRRC8 volume‐regulated anion channel inhibitor, DCPIB, inhibits mitochondrial respiration independently of the channel

et al. 2019

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There has been a resurgence of interest in the volume-regulated anion channel (VRAC) since the recent cloning of the LRRC8A-E gene family that encodes VRAC.The channel is a heteromer comprised of LRRC8A and at least one other family member; disruption of LRRC8A expression abolishes VRAC activity. The best-inclass VRAC inhibitor, DCPIB, suffers from off-target activity toward several different channels and transporters. Considering that some anion channel inhibitors also suppress mitochondrial respiration, we systematically explored whether DCPIB inhibits respiration in wild type (WT) and LRRC8A-knockout HAP-1 and HEK-293 cells. Knockout of LRRC8A had no apparent effects on cell morphology, proliferation rate, mitochondrial content, or expression of several mitochondrial genes in HAP-1 cells. Addition of 10 µM DCPIB, a concentration typically used to inhibit VRAC, suppressed basal and ATP-linked respiration in part through uncoupling the inner mitochondrial membrane (IMM) proton gradient and membrane potential.Additionally, DCPIB inhibits the activity of complex I, II, and III of the electron transport chain (ETC). Surprisingly, the effects of DCPIB on mitochondrial function are also observed in HAP-1 and HEK-293 cells which lack LRRC8A expression.Finally, we demonstrate that DCPIB activates ATP-inhibitable potassium channels comprised of heterologously expressed Kir6.2 and SUR1 subunits. These data indicate that DCPIB suppresses mitochondrial respiration and ATP production by dissipating the mitochondrial membrane potential and inhibiting complexes I-III of the ETC. They further justify the need for the development of sharper pharmacological tools for evaluating the integrative physiology and therapeutic potential of VRAC in human diseases. K E Y W O R D SDCPIB, LRRC8, mitochondria, respiration

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Section: Introductionmentioning

confidence: 54%

The LRRC8 volume‐regulated anion channel inhibitor, DCPIB, inhibits mitochondrial respiration independently of the channel

et al. 2019

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“…Overall, these findings suggest that deletion of Lrrc8a does not impact the numbers of neurons, astrocytes, oligodendrocytes, or the degree of myelination. The lack of changes in the CNS is somewhat surprising because VRAC was proposed to regulate cell proliferation, migration, differentiation, and survival in many cell types (reviews (Nilius et al 1997;Lang et al 1998;Hoffmann et al 2009) and recent experimental studies (Kumar et al 2014;Bao et al 2018;Chen et al 2019)).…”

Section: Lrrc8a Knockout Animals Display Astrogliosis But No Changesmentioning

confidence: 99%

“…Therefore, subsequent studies employed conditional, region-and cell type-specific Lrrc8a gene deletion. This approach revealed tissue-specific contributions of VRAC to reproductive function (spermatogenesis), regulation of adipocyte size, insulin signaling, glucose homeostasis, and innate immunity (Luck et al 2018;Bao et al 2018;Zhang et al 2017;Kang et al 2018;Zhou et al 2020). Although the central nervous system (CNS) has long been considered to be the tissue with the highest sensitivity to cell volume alterations and VRAC activity (Kimelberg and Mongin 1998;Kimelberg 2005;Mongin 2007;Mongin 2016;Wilson and Mongin 2018), few studies have analyzed the functional significance of LRRC8 proteins in the brain.…”

Section: Introductionmentioning

confidence: 99%

Late adolescence mortality in mice with brain-specific deletion of the volume-regulated anion channel subunit LRRC8A

Wilson

Dohare

Orbeta

et al. 2020

Preprint

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The leucine-rich repeat-containing family 8 member A (LRRC8A) is an essential subunit of the volume-regulated anion channel (VRAC). VRAC is indispensable for cell volume regulation but its broader physiological functions remain under investigation. Astrocyte-targeted Lrrc8a deletion in the nervous system reduces neuronal excitability, impairs synaptic plasticity and memory, and protects against ischemic damage. Here we show that deletion of LRRC8A in all brain cells, using Nestin Cre -driven Lrrc8a fl/fl excision, is lethal. Mice devoid of brain LRRC8A are born close to the expected Mendelian ratio and develop without overt histological abnormalities. Nevertheless, they all die between 5 to 8 weeks of age with a seizure-like phenotype. Consistent with seizures, we found disruptions in cell excitability, GABAergic signaling, and astrocytic production of the GABA precursor glutamine, all of which might contribute to mortality. This work provides the first evidence of a critical role for VRAC in control of brain excitability during maturation.

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“…EPB41 is one of the pleiotropic monogenic disorder's genes that cause male infertility, and manifests as azoospermia [39]. Bao et al demonstrated that LRRC8A-dependent VRAC activity is essential for male germ cell development and fertility [40]. LRRC8/VRAC anion channels are required for late stages of spermatid development in mice [41].…”

Section: Discussionmentioning

confidence: 99%

RNA-sequencing and bioinformatics analysis of long noncoding RNAs and mRNAs in the asthenozoospermia

Lü

Wang

et al. 2020

Bioscience Reports

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Asthenozoospermia is one of the major causes of human male infertility. Long non-coding RNAs (lncRNAs) play critical roles in the spermatogenesis processes. This study aims to investigate the intricate regulatory network associated with asthenozoospermia. The lncRNAs expression profile was analyzed in the asthenozoospermia seminal plasma exosomes by RNA-sequencing, and the functions of differentially expressed genes (DEGs) were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and DO (Disease Ontology) enrichment analyses. Pearson’s correlation test was utilized to calculate the correlation coefficients between lncRNA and mRNAs. Moreover, the lncRNA-miRNA-mRNA co-expression network was constructed with bioinformatics. From the co-expression analyses, we identified the cis regulated correlation pairs lncRNAs-mRNAs. To confirm sequencing results with five of the identified DElncRNAs were verified with quantitative reverse-transcription polymerase chain reaction (qRT-PCR). We identified 4,228 significantly DEGs, 995 known DElncRNAs, 2,338 DEmRNAs and 11,706 novel DElncRNAs between asthenozoospermia and normal group. GO and KEGG analyses showed that the DEGs were mainly associated with metabolism, transcription, ribosome and channel activity. We found 254,981 positive correlations lncRNA-mRNA pairs through correlation analysis. The detailed lncRNA-miRNA-mRNA regulatory network included 11 lncRNAs, 35 miRNAs and 59 mRNAs. From the co-expression analyses, we identified 7 cis-regulated correlation pair lncRNAs-mRNAs. Additionally, the qRT-PCR analysis confirmed our sequencing results. Our study constructed the lncRNA-mRNA-miRNA regulation networks in asthenozoospermia. Therefore, the study findings provide a set of pivotal lncRNAs for future investigation into the molecular mechanisms of asthenozoospermia.

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Deficient LRRC8A-dependent volume-regulated anion channel activity is associated with male infertility in mice

Cited by 29 publications

References 58 publications

The LRRC8 volume‐regulated anion channel inhibitor, DCPIB, inhibits mitochondrial respiration independently of the channel

The LRRC8 volume‐regulated anion channel inhibitor, DCPIB, inhibits mitochondrial respiration independently of the channel

Late adolescence mortality in mice with brain-specific deletion of the volume-regulated anion channel subunit LRRC8A

RNA-sequencing and bioinformatics analysis of long noncoding RNAs and mRNAs in the asthenozoospermia

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