The role of Sox6 in zebrafish muscle fiber type specification
Background: The transcription factor Sox6 has been implicated in regulating muscle fiber type-specific gene expression in mammals. In zebrafish, loss of function of the transcription factor Prdm1a results in a slow to fast-twitch fiber type transformation presaged by ectopic expression of sox6 in sl...
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2015
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| Online Access: | https://hdl.handle.net/10356/107147 http://hdl.handle.net/10220/25404 |
| _version_ | 1826125476471504896 |
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| author | Jackson, Harriet E Ono, Yosuke Wang, Xingang Elworthy, Stone Cunliffe, Vincent T Ingham, Philip William |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Jackson, Harriet E Ono, Yosuke Wang, Xingang Elworthy, Stone Cunliffe, Vincent T Ingham, Philip William |
| author_sort | Jackson, Harriet E |
| collection | NTU |
| description | Background: The transcription factor Sox6 has been implicated in regulating muscle fiber type-specific gene expression in mammals. In zebrafish, loss of function of the transcription factor Prdm1a results in a slow to fast-twitch fiber type transformation presaged by ectopic expression of sox6 in slow-twitch progenitors. Morpholino-mediated Sox6 knockdown can suppress this transformation but causes ectopic expression of only one of three slow-twitch specific genes assayed. Here, we use gain and loss of function analysis to analyse further the role of Sox6 in zebrafish muscle fiber type specification. Methods: The GAL4 binary misexpression system was used to express Sox6 ectopically in zebrafish embryos. Cis-regulatory elements were characterized using transgenic fish. Zinc finger nuclease mediated targeted mutagenesis was used to analyse the effects of loss of Sox6 function in embryonic, larval and adult zebrafish. Zebrafish transgenic for the GCaMP3 Calcium reporter were used to assay Ca2+ transients in wild-type and mutant muscle fibres. Results: Ectopic Sox6 expression is sufficient to downregulate slow-twitch specific gene expression in zebrafish embryos. Cis-regulatory elements upstream of the slow myosin heavy chain 1 (smyhc1) and slow troponin c (tnnc1b) genes contain putative Sox6 binding sites required for repression of the former but not the latter. Embryos homozygous for sox6 null alleles expressed tnnc1b throughout the fast-twitch muscle whereas other slow-specific muscle genes, including smyhc1, were expressed ectopically in only a subset of fast-twitch fibers. Ca2+ transients in sox6 mutant fast-twitch fibers were intermediate in their speed and amplitude between those of wild-type slow- and fast-twitch fibers. sox6 homozygotes survived to adulthood and exhibited continued misexpression of tnnc1b as well as smaller slow-twitch fibers. They also exhibited a striking curvature of the spine. Conclusions: The Sox6 transcription factor is a key regulator of fast-twitch muscle fiber differentiation in the zebrafish, a role similar to that ascribed to its murine ortholog. |
| first_indexed | 2024-10-01T06:37:16Z |
| format | Journal Article |
| id | ntu-10356/107147 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:37:16Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | ntu-10356/1071472022-02-16T16:31:06Z The role of Sox6 in zebrafish muscle fiber type specification Jackson, Harriet E Ono, Yosuke Wang, Xingang Elworthy, Stone Cunliffe, Vincent T Ingham, Philip William Lee Kong Chian School of Medicine (LKCMedicine) DRNTU::Science::Biological sciences::Zoology Background: The transcription factor Sox6 has been implicated in regulating muscle fiber type-specific gene expression in mammals. In zebrafish, loss of function of the transcription factor Prdm1a results in a slow to fast-twitch fiber type transformation presaged by ectopic expression of sox6 in slow-twitch progenitors. Morpholino-mediated Sox6 knockdown can suppress this transformation but causes ectopic expression of only one of three slow-twitch specific genes assayed. Here, we use gain and loss of function analysis to analyse further the role of Sox6 in zebrafish muscle fiber type specification. Methods: The GAL4 binary misexpression system was used to express Sox6 ectopically in zebrafish embryos. Cis-regulatory elements were characterized using transgenic fish. Zinc finger nuclease mediated targeted mutagenesis was used to analyse the effects of loss of Sox6 function in embryonic, larval and adult zebrafish. Zebrafish transgenic for the GCaMP3 Calcium reporter were used to assay Ca2+ transients in wild-type and mutant muscle fibres. Results: Ectopic Sox6 expression is sufficient to downregulate slow-twitch specific gene expression in zebrafish embryos. Cis-regulatory elements upstream of the slow myosin heavy chain 1 (smyhc1) and slow troponin c (tnnc1b) genes contain putative Sox6 binding sites required for repression of the former but not the latter. Embryos homozygous for sox6 null alleles expressed tnnc1b throughout the fast-twitch muscle whereas other slow-specific muscle genes, including smyhc1, were expressed ectopically in only a subset of fast-twitch fibers. Ca2+ transients in sox6 mutant fast-twitch fibers were intermediate in their speed and amplitude between those of wild-type slow- and fast-twitch fibers. sox6 homozygotes survived to adulthood and exhibited continued misexpression of tnnc1b as well as smaller slow-twitch fibers. They also exhibited a striking curvature of the spine. Conclusions: The Sox6 transcription factor is a key regulator of fast-twitch muscle fiber differentiation in the zebrafish, a role similar to that ascribed to its murine ortholog. Published version 2015-04-16T02:30:43Z 2019-12-06T22:25:42Z 2015-04-16T02:30:43Z 2019-12-06T22:25:42Z 2015 2015 Journal Article Jackson, H. E., Ono, Y., Wang, X., Elworthy, S., Cunliffe, V. T., & Ingham, P. W. (2015). The role of Sox6 in zebrafish muscle fiber type specification. Skeletal Muscle, 5. 2044-5040 https://hdl.handle.net/10356/107147 http://hdl.handle.net/10220/25404 10.1186/s13395-014-0026-2 25671076 en Skeletal muscle © 2015 Jackson et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Zoology Jackson, Harriet E Ono, Yosuke Wang, Xingang Elworthy, Stone Cunliffe, Vincent T Ingham, Philip William The role of Sox6 in zebrafish muscle fiber type specification |
| title | The role of Sox6 in zebrafish muscle fiber type specification |
| title_full | The role of Sox6 in zebrafish muscle fiber type specification |
| title_fullStr | The role of Sox6 in zebrafish muscle fiber type specification |
| title_full_unstemmed | The role of Sox6 in zebrafish muscle fiber type specification |
| title_short | The role of Sox6 in zebrafish muscle fiber type specification |
| title_sort | role of sox6 in zebrafish muscle fiber type specification |
| topic | DRNTU::Science::Biological sciences::Zoology |
| url | https://hdl.handle.net/10356/107147 http://hdl.handle.net/10220/25404 |
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