Inactivation of zebrafish mrf4 leads to myofibril misalignment and motor axon growth disorganization

Yun Hsin Wang, Chun Kai Li, Gang-Hui Lee, Huey Jen Tsay, Huai Jen Tsai, Yau Hung Chen

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Mrf4 is a basic helix-loop-helix (bHLH) transcription factor associated with myogenesis. Two mrf4 transcripts, mrf4_tv1 and mrf4_tv2, were identified in zebrafish generated by alternative splicing. To study their biological functions, we separately injected the Mrf4-morpholinos, including MO1 (mrf4_tv1:mrf4_tv2 knockdown), MO2+MO3 (mrf4_tv1:mrf4_tv2 knockdown), MO3 (mrf4_tv1 knockdown), and MO4 (mrf4_tv2 knockdown), into zebrafish embryos to observe mrf4 gene knockdown phenotypes. No phenotypic abnormalities were observed following injection with 0.5 ng of MO1 but those injected with 4.5, 9, or 13.5 ng displayed curved-body phenotypes, such as indistinct somite boundaries, and a lack of uniformly sized cell blocks. Similar results were also observed in the (MO2+MO3)-, MO3-, and MO4-injected groups. To further investigate the molecular mechanisms that lead to curved-body phenotypes, we stained embryos with α-bungrotoxin and specific monoclonal antibodies F59, Znp1, and Zn5 to detect morphological changes in acetyl-choline receptor (AChR) clusters, muscle fibers, common path of the primary neurons, and secondary neurons axonal projections, respectively. Our results show that the muscle fibers of mrf4_(tv1:tv2)-morphant aligned disorderly and lost their integrity and attachment, while the defects became milder in either mrf4_tv1-morphant or mrf4_tv2-morphant. On the other hand, reduced axonal projections and AChR clusters were found in both mrf4_tv2-morphant and mrf4_(tv1:tv2)-morphant but distributed normally in the mrf4_tv1-morphant. We conclude that Mrf4_tv2 is involved in alignment of muscle fibers, and Mrf4_tv1 might have cooperative function with Mrf4_tv2 in muscle fiber alignment, without affecting the muscle-nerve connection.

Original languageEnglish
Pages (from-to)1043-1050
Number of pages8
JournalDevelopmental Dynamics
Volume237
Issue number4
DOIs
Publication statusPublished - 2008 Apr 1

Fingerprint

Myofibrils
Zebrafish
Axons
Muscles
Growth
Choline
Phenotype
Embryonic Structures
Gene Knockdown Techniques
Basic Helix-Loop-Helix Transcription Factors
Neurons
Morpholinos
Somites
Muscle Development
Alternative Splicing
Monoclonal Antibodies
Injections

All Science Journal Classification (ASJC) codes

  • Developmental Biology

Cite this

Wang, Yun Hsin ; Li, Chun Kai ; Lee, Gang-Hui ; Tsay, Huey Jen ; Tsai, Huai Jen ; Chen, Yau Hung. / Inactivation of zebrafish mrf4 leads to myofibril misalignment and motor axon growth disorganization. In: Developmental Dynamics. 2008 ; Vol. 237, No. 4. pp. 1043-1050.
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abstract = "Mrf4 is a basic helix-loop-helix (bHLH) transcription factor associated with myogenesis. Two mrf4 transcripts, mrf4_tv1 and mrf4_tv2, were identified in zebrafish generated by alternative splicing. To study their biological functions, we separately injected the Mrf4-morpholinos, including MO1 (mrf4_tv1:mrf4_tv2 knockdown), MO2+MO3 (mrf4_tv1:mrf4_tv2 knockdown), MO3 (mrf4_tv1 knockdown), and MO4 (mrf4_tv2 knockdown), into zebrafish embryos to observe mrf4 gene knockdown phenotypes. No phenotypic abnormalities were observed following injection with 0.5 ng of MO1 but those injected with 4.5, 9, or 13.5 ng displayed curved-body phenotypes, such as indistinct somite boundaries, and a lack of uniformly sized cell blocks. Similar results were also observed in the (MO2+MO3)-, MO3-, and MO4-injected groups. To further investigate the molecular mechanisms that lead to curved-body phenotypes, we stained embryos with α-bungrotoxin and specific monoclonal antibodies F59, Znp1, and Zn5 to detect morphological changes in acetyl-choline receptor (AChR) clusters, muscle fibers, common path of the primary neurons, and secondary neurons axonal projections, respectively. Our results show that the muscle fibers of mrf4_(tv1:tv2)-morphant aligned disorderly and lost their integrity and attachment, while the defects became milder in either mrf4_tv1-morphant or mrf4_tv2-morphant. On the other hand, reduced axonal projections and AChR clusters were found in both mrf4_tv2-morphant and mrf4_(tv1:tv2)-morphant but distributed normally in the mrf4_tv1-morphant. We conclude that Mrf4_tv2 is involved in alignment of muscle fibers, and Mrf4_tv1 might have cooperative function with Mrf4_tv2 in muscle fiber alignment, without affecting the muscle-nerve connection.",
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Inactivation of zebrafish mrf4 leads to myofibril misalignment and motor axon growth disorganization. / Wang, Yun Hsin; Li, Chun Kai; Lee, Gang-Hui; Tsay, Huey Jen; Tsai, Huai Jen; Chen, Yau Hung.

In: Developmental Dynamics, Vol. 237, No. 4, 01.04.2008, p. 1043-1050.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Inactivation of zebrafish mrf4 leads to myofibril misalignment and motor axon growth disorganization

AU - Wang, Yun Hsin

AU - Li, Chun Kai

AU - Lee, Gang-Hui

AU - Tsay, Huey Jen

AU - Tsai, Huai Jen

AU - Chen, Yau Hung

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N2 - Mrf4 is a basic helix-loop-helix (bHLH) transcription factor associated with myogenesis. Two mrf4 transcripts, mrf4_tv1 and mrf4_tv2, were identified in zebrafish generated by alternative splicing. To study their biological functions, we separately injected the Mrf4-morpholinos, including MO1 (mrf4_tv1:mrf4_tv2 knockdown), MO2+MO3 (mrf4_tv1:mrf4_tv2 knockdown), MO3 (mrf4_tv1 knockdown), and MO4 (mrf4_tv2 knockdown), into zebrafish embryos to observe mrf4 gene knockdown phenotypes. No phenotypic abnormalities were observed following injection with 0.5 ng of MO1 but those injected with 4.5, 9, or 13.5 ng displayed curved-body phenotypes, such as indistinct somite boundaries, and a lack of uniformly sized cell blocks. Similar results were also observed in the (MO2+MO3)-, MO3-, and MO4-injected groups. To further investigate the molecular mechanisms that lead to curved-body phenotypes, we stained embryos with α-bungrotoxin and specific monoclonal antibodies F59, Znp1, and Zn5 to detect morphological changes in acetyl-choline receptor (AChR) clusters, muscle fibers, common path of the primary neurons, and secondary neurons axonal projections, respectively. Our results show that the muscle fibers of mrf4_(tv1:tv2)-morphant aligned disorderly and lost their integrity and attachment, while the defects became milder in either mrf4_tv1-morphant or mrf4_tv2-morphant. On the other hand, reduced axonal projections and AChR clusters were found in both mrf4_tv2-morphant and mrf4_(tv1:tv2)-morphant but distributed normally in the mrf4_tv1-morphant. We conclude that Mrf4_tv2 is involved in alignment of muscle fibers, and Mrf4_tv1 might have cooperative function with Mrf4_tv2 in muscle fiber alignment, without affecting the muscle-nerve connection.

AB - Mrf4 is a basic helix-loop-helix (bHLH) transcription factor associated with myogenesis. Two mrf4 transcripts, mrf4_tv1 and mrf4_tv2, were identified in zebrafish generated by alternative splicing. To study their biological functions, we separately injected the Mrf4-morpholinos, including MO1 (mrf4_tv1:mrf4_tv2 knockdown), MO2+MO3 (mrf4_tv1:mrf4_tv2 knockdown), MO3 (mrf4_tv1 knockdown), and MO4 (mrf4_tv2 knockdown), into zebrafish embryos to observe mrf4 gene knockdown phenotypes. No phenotypic abnormalities were observed following injection with 0.5 ng of MO1 but those injected with 4.5, 9, or 13.5 ng displayed curved-body phenotypes, such as indistinct somite boundaries, and a lack of uniformly sized cell blocks. Similar results were also observed in the (MO2+MO3)-, MO3-, and MO4-injected groups. To further investigate the molecular mechanisms that lead to curved-body phenotypes, we stained embryos with α-bungrotoxin and specific monoclonal antibodies F59, Znp1, and Zn5 to detect morphological changes in acetyl-choline receptor (AChR) clusters, muscle fibers, common path of the primary neurons, and secondary neurons axonal projections, respectively. Our results show that the muscle fibers of mrf4_(tv1:tv2)-morphant aligned disorderly and lost their integrity and attachment, while the defects became milder in either mrf4_tv1-morphant or mrf4_tv2-morphant. On the other hand, reduced axonal projections and AChR clusters were found in both mrf4_tv2-morphant and mrf4_(tv1:tv2)-morphant but distributed normally in the mrf4_tv1-morphant. We conclude that Mrf4_tv2 is involved in alignment of muscle fibers, and Mrf4_tv1 might have cooperative function with Mrf4_tv2 in muscle fiber alignment, without affecting the muscle-nerve connection.

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