{"doi":"10.3389/fphys.2020.00326","title":"Patch-Clamp Analysis of the Mitochondrial H+ Leak in Brown and Beige Fat","abstract":"Mitochondria convert the energy of substrate oxidation into adenosine triphosphate (ATP) and heat. Although ATP production has become a focal point of research in bioenergetics, mitochondrial thermogenesis is also crucial for energy metabolism. Mitochondria generate heat due to H+ leak across the inner mitochondrial membrane which is mediated by mitochondrial uncoupling proteins. The mitochondrial H+ leak was first identified, and studied for many decades, using mitochondrial respiration technique. Unfortunately, this method measures H+ leak indirectly, and its precision is insufficient for the rigorous insight into the mitochondrial function at the molecular level. Direct patch-clamp recording of H+ leak would have a significantly higher amplitude and time resolution, but application of the patch-clamp technique to a small subcellular organelle such as mitochondria has been challenging. We developed a method that facilitates patch-clamp recording from the whole inner mitochondrial membrane, enabling the direct measurement of small H+ leak currents via uncoupling proteins and thus, providing a rigorous understanding of the molecular mechanisms involved. In this paper we cover the methodology of measuring the H+ leak in mitochondria of specialized thermogenic tissues brown and beige fat.","journal":"Frontiers in Physiology","year":2020,"id":89917,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9554,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":453660,"name":"Yuriy Kirichok","orcid":"0000-0001-7155-843X","position":1,"is_corresponding":false},{"id":454265,"name":"Ambre M. Bertholet","orcid":null,"position":0,"is_corresponding":true}],"reference_count":46,"raw_metadata":null,"created_at":"2026-07-18T22:02:27.195966Z","pmid":"32351404","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}