Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development
The development of craniofacial skeletal structures requires well-orchestrated tissue interactions controlled by distinct molecular signals. Disruptions in normal function of these molecular signals have been associated with a wide range of craniofacial malformations. A pathway mediated by estrogens...
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PeerJ Inc.
2016-03-01
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author | Ehsan Pashay Ahi Benjamin S. Walker Christopher S. Lassiter Zophonías O. Jónsson |
author_facet | Ehsan Pashay Ahi Benjamin S. Walker Christopher S. Lassiter Zophonías O. Jónsson |
author_sort | Ehsan Pashay Ahi |
collection | DOAJ |
description | The development of craniofacial skeletal structures requires well-orchestrated tissue interactions controlled by distinct molecular signals. Disruptions in normal function of these molecular signals have been associated with a wide range of craniofacial malformations. A pathway mediated by estrogens is one of those molecular signals that plays role in formation of bone and cartilage including craniofacial skeletogenesis. Studies in zebrafish have shown that while higher concentrations of 17-β estradiol (E2) cause severe craniofacial defects, treatment with lower concentrations result in subtle changes in head morphology characterized with shorter snouts and flatter faces. The molecular basis for these morphological changes, particularly the subtle skeletal effects mediated by lower E2 concentrations, remains unexplored. In the present study we address these effects at a molecular level by quantitative expression analysis of sets of candidate genes in developing heads of zebrafish larvae treated with two different E2 concentrations. To this end, we first validated three suitable reference genes, ppia2, rpl8 and tbp, to permit sensitive quantitative real-time PCR analysis. Next, we profiled the expression of 28 skeletogenesis-associated genes that potentially respond to estrogen signals and play role in craniofacial development. We found E2 mediated differential expression of genes involved in extracellular matrix (ECM) remodelling, mmp2/9/13, sparc and timp2a, as well as components of skeletogenic pathways, bmp2a, erf, ptch1/2, rankl, rarab and sfrp1a. Furthermore, we identified a co-expressed network of genes, including cpn1, dnajc3, esr1, lman1, rrbp1a, ssr1 and tram1 with a stronger inductive response to a lower dose of E2 during larval head development. |
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spelling | doaj.art-29f52eb8b6f14171bf874872dae42a8c2023-12-02T21:59:54ZengPeerJ Inc.PeerJ2167-83592016-03-014e187810.7717/peerj.1878Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head developmentEhsan Pashay Ahi0Benjamin S. Walker1Christopher S. Lassiter2Zophonías O. Jónsson3Institute of Life and Environmental Sciences, University of Iceland, Reykjavik, IcelandBiology Department, Roanoke College, Salem, VA, United StatesBiology Department, Roanoke College, Salem, VA, United StatesInstitute of Life and Environmental Sciences, University of Iceland, Reykjavik, IcelandThe development of craniofacial skeletal structures requires well-orchestrated tissue interactions controlled by distinct molecular signals. Disruptions in normal function of these molecular signals have been associated with a wide range of craniofacial malformations. A pathway mediated by estrogens is one of those molecular signals that plays role in formation of bone and cartilage including craniofacial skeletogenesis. Studies in zebrafish have shown that while higher concentrations of 17-β estradiol (E2) cause severe craniofacial defects, treatment with lower concentrations result in subtle changes in head morphology characterized with shorter snouts and flatter faces. The molecular basis for these morphological changes, particularly the subtle skeletal effects mediated by lower E2 concentrations, remains unexplored. In the present study we address these effects at a molecular level by quantitative expression analysis of sets of candidate genes in developing heads of zebrafish larvae treated with two different E2 concentrations. To this end, we first validated three suitable reference genes, ppia2, rpl8 and tbp, to permit sensitive quantitative real-time PCR analysis. Next, we profiled the expression of 28 skeletogenesis-associated genes that potentially respond to estrogen signals and play role in craniofacial development. We found E2 mediated differential expression of genes involved in extracellular matrix (ECM) remodelling, mmp2/9/13, sparc and timp2a, as well as components of skeletogenic pathways, bmp2a, erf, ptch1/2, rankl, rarab and sfrp1a. Furthermore, we identified a co-expressed network of genes, including cpn1, dnajc3, esr1, lman1, rrbp1a, ssr1 and tram1 with a stronger inductive response to a lower dose of E2 during larval head development.https://peerj.com/articles/1878.pdfCraniofacial skeletonDevelopmentEstrogenGene expressionZebrafish larvaeqPCR |
spellingShingle | Ehsan Pashay Ahi Benjamin S. Walker Christopher S. Lassiter Zophonías O. Jónsson Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development PeerJ Craniofacial skeleton Development Estrogen Gene expression Zebrafish larvae qPCR |
title | Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development |
title_full | Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development |
title_fullStr | Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development |
title_full_unstemmed | Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development |
title_short | Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development |
title_sort | investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development |
topic | Craniofacial skeleton Development Estrogen Gene expression Zebrafish larvae qPCR |
url | https://peerj.com/articles/1878.pdf |
work_keys_str_mv | AT ehsanpashayahi investigationoftheeffectsofestrogenonskeletalgeneexpressionduringzebrafishlarvalheaddevelopment AT benjaminswalker investigationoftheeffectsofestrogenonskeletalgeneexpressionduringzebrafishlarvalheaddevelopment AT christopherslassiter investigationoftheeffectsofestrogenonskeletalgeneexpressionduringzebrafishlarvalheaddevelopment AT zophoniasojonsson investigationoftheeffectsofestrogenonskeletalgeneexpressionduringzebrafishlarvalheaddevelopment |