In vivo myosin step-size from zebrafish skeletal muscle
Muscle myosins transduce ATP free energy into actin displacement to power contraction. In vivo, myosin side chains are modified post-translationally under native conditions, potentially impacting function. Single myosin detection provides the ‘bottom-up’ myosin characterization probing basic mechani...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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The Royal Society
2016-01-01
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| Series: | Open Biology |
| Subjects: | |
| Online Access: | https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.160075 |
| _version_ | 1828885960321925120 |
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| author | Thomas P. Burghardt Katalin Ajtai Xiaojing Sun Naoko Takubo Yihua Wang |
| author_facet | Thomas P. Burghardt Katalin Ajtai Xiaojing Sun Naoko Takubo Yihua Wang |
| author_sort | Thomas P. Burghardt |
| collection | DOAJ |
| description | Muscle myosins transduce ATP free energy into actin displacement to power contraction. In vivo, myosin side chains are modified post-translationally under native conditions, potentially impacting function. Single myosin detection provides the ‘bottom-up’ myosin characterization probing basic mechanisms without ambiguities inherent to ensemble observation. Macroscopic muscle physiological experimentation provides the definitive ‘top-down’ phenotype characterizations that are the concerns in translational medicine. In vivo single myosin detection in muscle from zebrafish embryo models for human muscle fulfils ambitions for both bottom-up and top-down experimentation. A photoactivatable green fluorescent protein (GFP)-tagged myosin light chain expressed in transgenic zebrafish skeletal muscle specifically modifies the myosin lever-arm. Strychnine induces the simultaneous contraction of the bilateral tail muscles in a live embryo, causing them to be isometric while active. Highly inclined thin illumination excites the GFP tag of single lever-arms and its super-resolution orientation is measured from an active isometric muscle over a time sequence covering many transduction cycles. Consecutive frame lever-arm angular displacement converts to step-size by its product with the estimated lever-arm length. About 17% of the active myosin steps that fall between 2 and 7 nm are implicated as powerstrokes because they are beyond displacements detected from either relaxed or ATP-depleted (rigor) muscle. |
| first_indexed | 2024-12-13T11:36:01Z |
| format | Article |
| id | doaj.art-28dd798b7325493ea5edd0f0f67bd7dc |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-12-13T11:36:01Z |
| publishDate | 2016-01-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-28dd798b7325493ea5edd0f0f67bd7dc2022-12-21T23:47:46ZengThe Royal SocietyOpen Biology2046-24412016-01-016510.1098/rsob.160075160075In vivo myosin step-size from zebrafish skeletal muscleThomas P. BurghardtKatalin AjtaiXiaojing SunNaoko TakuboYihua WangMuscle myosins transduce ATP free energy into actin displacement to power contraction. In vivo, myosin side chains are modified post-translationally under native conditions, potentially impacting function. Single myosin detection provides the ‘bottom-up’ myosin characterization probing basic mechanisms without ambiguities inherent to ensemble observation. Macroscopic muscle physiological experimentation provides the definitive ‘top-down’ phenotype characterizations that are the concerns in translational medicine. In vivo single myosin detection in muscle from zebrafish embryo models for human muscle fulfils ambitions for both bottom-up and top-down experimentation. A photoactivatable green fluorescent protein (GFP)-tagged myosin light chain expressed in transgenic zebrafish skeletal muscle specifically modifies the myosin lever-arm. Strychnine induces the simultaneous contraction of the bilateral tail muscles in a live embryo, causing them to be isometric while active. Highly inclined thin illumination excites the GFP tag of single lever-arms and its super-resolution orientation is measured from an active isometric muscle over a time sequence covering many transduction cycles. Consecutive frame lever-arm angular displacement converts to step-size by its product with the estimated lever-arm length. About 17% of the active myosin steps that fall between 2 and 7 nm are implicated as powerstrokes because they are beyond displacements detected from either relaxed or ATP-depleted (rigor) muscle.https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.160075single myosin detection in vivohighly inclined thin illuminationtransgenic zebrafish skeletal musclestrychnine induced contractionzebrafish skeletal myosin powerstrokezebrafish skeletal myosin step-size |
| spellingShingle | Thomas P. Burghardt Katalin Ajtai Xiaojing Sun Naoko Takubo Yihua Wang In vivo myosin step-size from zebrafish skeletal muscle Open Biology single myosin detection in vivo highly inclined thin illumination transgenic zebrafish skeletal muscle strychnine induced contraction zebrafish skeletal myosin powerstroke zebrafish skeletal myosin step-size |
| title | In vivo myosin step-size from zebrafish skeletal muscle |
| title_full | In vivo myosin step-size from zebrafish skeletal muscle |
| title_fullStr | In vivo myosin step-size from zebrafish skeletal muscle |
| title_full_unstemmed | In vivo myosin step-size from zebrafish skeletal muscle |
| title_short | In vivo myosin step-size from zebrafish skeletal muscle |
| title_sort | in vivo myosin step size from zebrafish skeletal muscle |
| topic | single myosin detection in vivo highly inclined thin illumination transgenic zebrafish skeletal muscle strychnine induced contraction zebrafish skeletal myosin powerstroke zebrafish skeletal myosin step-size |
| url | https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.160075 |
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