{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T22:32:04Z","timestamp":1788474724713,"version":"build-2803163510"},"reference-count":51,"publisher":"American Physiological Society","issue":"4","funder":[{"name":"AHA","award":["18CDA34110044"],"award-info":[{"award-number":["18CDA34110044"]}]},{"name":"NIH","award":["R01 HL130318-01"],"award-info":[{"award-number":["R01 HL130318-01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["American Journal of Physiology-Cell Physiology"],"published-print":{"date-parts":[[2019,10,1]]},"abstract":"<jats:p> Diaphragm abnormalities in aging or chronic diseases include impaired mitochondrial respiration and H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> emission, which can be measured using saponin-permeabilized muscle fibers. Mouse diaphragm presents a challenge for isolation of fibers due to relatively high abundance of connective tissue in healthy muscle that is exacerbated in disease states. We tested a new approach to process mouse diaphragm for assessment of intact mitochondria respiration and ROS emission in saponin-permeabilized fibers. We used the red gastrocnemius (RG) as \u201cstandard\u201d limb muscle. Markers of mitochondrial content were two\u2013 to fourfold higher in diaphragm (Dia) than in RG ( P &lt; 0.05). Maximal O<jats:sub>2<\/jats:sub> consumption ( JO<jats:sub>2<\/jats:sub>: pmol\u00b7s<jats:sup>\u22121<\/jats:sup>\u00b7mg<jats:sup>\u22121<\/jats:sup>) in Dia was higher with glutamate, malate, and succinate (Dia 399\u2009\u00b1\u2009127, RG 148\u2009\u00b1\u200960; P &lt; 0.05) and palmitoyl-CoA + carnitine (Dia 15\u2009\u00b1\u20095, RG 7\u2009\u00b1\u20091; P &lt; 0.05) than in RG, but not different between muscles when JO<jats:sub>2<\/jats:sub> was normalized to citrate synthase activity. Absolute JO<jats:sub>2<\/jats:sub> for Dia was two\u2013 to fourfold higher than reported in previous studies. Mitochondrial JH<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> was higher in Dia than in RG ( P &lt; 0.05), but lower in Dia than in RG when JH<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> was normalized to citrate synthase activity. Our findings are consistent with an optimized diaphragm preparation for assessment of intact mitochondria in permeabilized fiber bundles. The data also suggest that higher mitochondrial content potentially makes the diaphragm more susceptible to \u201cmitochondrial onset\u201d myopathy. Overall, the new approach will facilitate testing and understanding of diaphragm mitochondrial function in mouse models that are used to advance biomedical research and human health. <\/jats:p>","DOI":"10.1152\/ajpcell.00184.2019","type":"journal-article","created":{"date-parts":[[2019,7,17]],"date-time":"2019-07-17T19:03:51Z","timestamp":1563390231000},"page":"C665-C673","source":"Crossref","is-referenced-by-count":10,"title":["Mitochondrial respiration and H<sub>2<\/sub>O<sub>2<\/sub> emission in saponin-permeabilized murine diaphragm fibers: optimization of fiber separation and comparison to limb muscle"],"prefix":"10.1152","volume":"317","author":[{"given":"Dongwoo","family":"Hahn","sequence":"first","affiliation":[{"name":"Department of Applied Physiology and Kinesiology, College of Health and Human Performance, University of Florida, Gainesville, Florida"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ravi 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