Effects of insulin like growth factors on early embryonic chick limb myogenesis

Limb muscles derive from pax3 expressing precursor cells that migrate from the hypaxial somite into the developing limb bud. Once there they begin to differentiate and express muscle determination genes such as MyoD. This process is regulated by a combination of inductive or inhibitory signals inclu...

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Main Authors: Mohammed, Rabeea Hazim, Anderton, Helen, Brameld, John Michael, Sweetman, Dylan
Format: Article
Published: Public Library of Science 2017
Online Access:https://eprints.nottingham.ac.uk/46986/
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author Mohammed, Rabeea Hazim
Anderton, Helen
Brameld, John Michael
Sweetman, Dylan
author_facet Mohammed, Rabeea Hazim
Anderton, Helen
Brameld, John Michael
Sweetman, Dylan
author_sort Mohammed, Rabeea Hazim
building Nottingham Research Data Repository
collection Online Access
description Limb muscles derive from pax3 expressing precursor cells that migrate from the hypaxial somite into the developing limb bud. Once there they begin to differentiate and express muscle determination genes such as MyoD. This process is regulated by a combination of inductive or inhibitory signals including Fgf18, retinoic acid, HGF, Notch and IGFs. IGFs are well known to affect late stages of muscle development and to promote both proliferation and differentiation. We examined their roles in early stage limb bud myogenesis using chicken embryos as an experimental model. Grafting beads soaked in purified recombinant IGF-I, IGF-II or small molecule inhibitors of specific signaling pathways into developing chick embryo limbs showed that both IGF-I and IGF-II induce expression of the early stage myogenic markers pax3 and MyoD as well as myogenin. Their effects on pax3 and MyoD expression were blocked by inhibitors of both the IGF type I receptor (picropodophyllotoxin, PPP) and MEK (U0126). The PI3K inhibitor LY294002 blocked IGF-II, but not IGF-I, induction of pax3 mRNA as well as the IGF-I, but not IGF-II, induction of MyoD mRNA. In addition SU5402, an FGFR/ VEGFR inhibitor, blocked the induction of MyoD by both IGFs but had no effect on pax3 induction, suggesting a role for FGF or VEGF signaling in their induction of MyoD. This was confirmed by in situ hybridization showing that FGF18, a known regulator of MyoD in limb myoblasts, was induced by IGF-I. In addition to their well-known effects on later stages of myogenesis via their induction of myogenin expression, both IGF-I and IGF-II induced pax3 and MyoD expression in developing chick embryos, indicating that they also regulate early stages of myogenesis. The data suggests that the IGFs may have slightly different effects on IGF1R signal transduction via PI3K and that their stimulatory effects on MyoD expression may be indirect, possibly via induction of FGF18 expression.
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spelling nottingham-469862020-05-04T19:10:23Z https://eprints.nottingham.ac.uk/46986/ Effects of insulin like growth factors on early embryonic chick limb myogenesis Mohammed, Rabeea Hazim Anderton, Helen Brameld, John Michael Sweetman, Dylan Limb muscles derive from pax3 expressing precursor cells that migrate from the hypaxial somite into the developing limb bud. Once there they begin to differentiate and express muscle determination genes such as MyoD. This process is regulated by a combination of inductive or inhibitory signals including Fgf18, retinoic acid, HGF, Notch and IGFs. IGFs are well known to affect late stages of muscle development and to promote both proliferation and differentiation. We examined their roles in early stage limb bud myogenesis using chicken embryos as an experimental model. Grafting beads soaked in purified recombinant IGF-I, IGF-II or small molecule inhibitors of specific signaling pathways into developing chick embryo limbs showed that both IGF-I and IGF-II induce expression of the early stage myogenic markers pax3 and MyoD as well as myogenin. Their effects on pax3 and MyoD expression were blocked by inhibitors of both the IGF type I receptor (picropodophyllotoxin, PPP) and MEK (U0126). The PI3K inhibitor LY294002 blocked IGF-II, but not IGF-I, induction of pax3 mRNA as well as the IGF-I, but not IGF-II, induction of MyoD mRNA. In addition SU5402, an FGFR/ VEGFR inhibitor, blocked the induction of MyoD by both IGFs but had no effect on pax3 induction, suggesting a role for FGF or VEGF signaling in their induction of MyoD. This was confirmed by in situ hybridization showing that FGF18, a known regulator of MyoD in limb myoblasts, was induced by IGF-I. In addition to their well-known effects on later stages of myogenesis via their induction of myogenin expression, both IGF-I and IGF-II induced pax3 and MyoD expression in developing chick embryos, indicating that they also regulate early stages of myogenesis. The data suggests that the IGFs may have slightly different effects on IGF1R signal transduction via PI3K and that their stimulatory effects on MyoD expression may be indirect, possibly via induction of FGF18 expression. Public Library of Science 2017-10-03 Article PeerReviewed Mohammed, Rabeea Hazim, Anderton, Helen, Brameld, John Michael and Sweetman, Dylan (2017) Effects of insulin like growth factors on early embryonic chick limb myogenesis. PLoS ONE, 12 (10). e0185775. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0185775 doi:10.1371/journal.pone.0185775 doi:10.1371/journal.pone.0185775
spellingShingle Mohammed, Rabeea Hazim
Anderton, Helen
Brameld, John Michael
Sweetman, Dylan
Effects of insulin like growth factors on early embryonic chick limb myogenesis
title Effects of insulin like growth factors on early embryonic chick limb myogenesis
title_full Effects of insulin like growth factors on early embryonic chick limb myogenesis
title_fullStr Effects of insulin like growth factors on early embryonic chick limb myogenesis
title_full_unstemmed Effects of insulin like growth factors on early embryonic chick limb myogenesis
title_short Effects of insulin like growth factors on early embryonic chick limb myogenesis
title_sort effects of insulin like growth factors on early embryonic chick limb myogenesis
url https://eprints.nottingham.ac.uk/46986/
https://eprints.nottingham.ac.uk/46986/
https://eprints.nottingham.ac.uk/46986/