{
  "abstract": "Neuronal intracellular chloride concentration shapes the polarity and strength of GABAA receptor-mediated inhibition. ClC-2 is a hyperpolarization-activated chloride channel proposed to support chloride extrusion, but its acute contribution to EGABA and neuronal chloride homeostasis remains incompletely understood. Here, we used the selective ClC-2 inhibitor AK-42 to test how acute ClC-2 inhibition affects GABAergic chloride signaling in organotypic hippocampal slice cultures. We combined targeted single-interneuron optogenetic activation of stCoChR-expressing GABAergic interneurons with gramicidin-perforated patch recordings to measure optogenetically evoked inhibitory postsynaptic currents and calculate EGABA and we used SuperClomeleon imaging to measure neuronal intracellular chloride. AK-42 depolarized EGABA, with a larger effect when stimulation was delivered 2 s rather than 100 ms after voltage-step onset. This delay-dependent effect is consistent with greater engagement of voltage- and time-dependent ClC-2 activity during prolonged voltage steps. AK-42 also abolished the more hyperpolarized EGABA normally measured at the 2-s stimulation delay and shifted baseline holding currents at hyperpolarized command potentials without measurably changing estimated resting membrane potential. SuperClomeleon imaging showed that AK-42 increased neuronal intracellular chloride in the presence of TTX, supporting a role for ClC-2 in basal chloride regulation. Together, these findings show that acute ClC-2 inhibition depolarizes EGABA, increases neuronal intracellular chloride, and alters the temporal dynamics of GABAergic chloride signaling, identifying ClC-2 as an acute regulator of synaptic GABAergic chloride signaling.",
  "abstract_shown": true,
  "affiliations": [
    "Harvard Medical School, Massachusetts General Hospital"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
      "name": "F. H. Shiu"
    },
    {
      "name": "V. I. Dzhala"
    },
    {
      "name": "K. Lillis"
    },
    {
      "affs": [
        0
      ],
      "name": "K. J. Staley"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/b0n6k9jkaahy2.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.09.24.753897",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.09.24.753897v1.full",
  "id": "b0n6k9jkaahy2",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.09.24.753897v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "neuroscience"
    ],
    [
      "openalex",
      "2804|Neuroscience and Neuropharmacology Research"
    ]
  ],
  "openalex": "W7214933300",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.09.24.753897v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-24",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/b0n6k9jkaahy2.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:25:29Z",
      "native_id": "10.64898/2026.09.24.753897",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.09.24.753897v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:neuroscience",
      "subfield": "neuroscience"
    }
  ],
  "title": "Acute ClC-2 blockade increases neuronal intracellular chloride and depolarizes EGABA",
  "title_text": "Acute ClC-2 blockade increases neuronal intracellular chloride and depolarizes EGABA",
  "type": "preprint",
  "updated_at": "2026-10-30T12:25:29Z",
  "url": "https://research.draconitic.com/paper/b0n6k9jkaahy2",
  "venue": "bioRxiv",
  "versions": []
}