Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules
IntroductionIn vertebrate limb morphogenesis, wingless-related integration site (Wnt) proteins and fibroblast growth factors (Fgfs) secreted from the apical ectodermal ridge (AER) coordinate proximodistal outgrowth. Fgfs also sustain sonic hedgehog (Shh) in the zone of polarizing activity (ZPA). Shh...
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Frontiers Media S.A.
2025-02-01
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| author | Jessica C. Britton Anett Somogyi-Leatigaga Billy A. Watson Endika Haro Cassidy G. Mulder Kari D. Kennedy Allen M. Cooper Kristen L. Whitley Ruth-Love Yeboah Jeanyoung Kim Micah C. Yu Micah C. Yu Jairo D. Campos Jairo D. Campos Japhet Amoah Japhet Amoah Shimako Kawauchi Eunyoung Kim Charmaine U. Pira Kerby C. Oberg |
| author_facet | Jessica C. Britton Anett Somogyi-Leatigaga Billy A. Watson Endika Haro Cassidy G. Mulder Kari D. Kennedy Allen M. Cooper Kristen L. Whitley Ruth-Love Yeboah Jeanyoung Kim Micah C. Yu Micah C. Yu Jairo D. Campos Jairo D. Campos Japhet Amoah Japhet Amoah Shimako Kawauchi Eunyoung Kim Charmaine U. Pira Kerby C. Oberg |
| author_sort | Jessica C. Britton |
| collection | DOAJ |
| description | IntroductionIn vertebrate limb morphogenesis, wingless-related integration site (Wnt) proteins and fibroblast growth factors (Fgfs) secreted from the apical ectodermal ridge (AER) coordinate proximodistal outgrowth. Fgfs also sustain sonic hedgehog (Shh) in the zone of polarizing activity (ZPA). Shh directs anteroposterior patterning and expansion and regulates AER-Fgfs, establishing a positive regulatory feedback loop that is vital in sustaining limb outgrowth. The transcription factor LIM homeodomain 2 (Lhx2) is expressed in the distal mesoderm and coordinates AER and ZPA signals that control cellular proliferation, differentiation, and shaping of the developing limb. Yet how Lhx2 is transcriptionally regulated to support such functions has only been partially characterized.Methods/ResultsWe have identified two limb-specific cis-regulatory modules (CRMs) active within the Lhx2 expression domain in the limb. Chromatin conformation analysis of the Lhx2 locus in mouse embryonic limb bud cells predicted CRMs-Lhx2 promoter interactions. Single-cell RNA-sequencing analysis of limb bud cells revealed co-expression of several Fgf-related Ets and Wnt-related Tcf/Lef transcripts in Lhx2-expressing cells. Additionally, disruption of Ets and Tcf/Lef binding sites resulted in loss of reporter-driven CRM activity. Finally, binding of β-catenin to both Lhx2-associated CRMs supports the associated binding of Tcf/Lef transcription factors.DiscussionThese results suggest a role for Ets and Tcf/Lef transcription factors in the regulation of Lhx2 expression through these limb-specific Lhx2-associated CRMs. Moreover, these CRMs provide a mechanism for Fgf and Wnt signaling to localize and maintain distal Lhx2 expression during vertebrate limb development. |
| format | Article |
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| institution | DOAJ |
| issn | 2296-634X |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
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| spelling | doaj-art-63f1dee5b3bb4546a121613e6f1ff0352025-08-20T03:00:50ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-02-011310.3389/fcell.2025.15527161552716Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modulesJessica C. Britton0Anett Somogyi-Leatigaga1Billy A. Watson2Endika Haro3Cassidy G. Mulder4Kari D. Kennedy5Allen M. Cooper6Kristen L. Whitley7Ruth-Love Yeboah8Jeanyoung Kim9Micah C. Yu10Micah C. Yu11Jairo D. Campos12Jairo D. Campos13Japhet Amoah14Japhet Amoah15Shimako Kawauchi16Eunyoung Kim17Charmaine U. Pira18Kerby C. Oberg19Department of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesSchool of Medicine, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesSchool of Medicine, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesSchool of Medicine, Loma Linda University, Loma Linda, CA, United StatesUC Irvine Transgenic Mouse Facility, University of Irvine, Irvine, CA, United StatesUC Irvine Transgenic Mouse Facility, University of Irvine, Irvine, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA, United StatesIntroductionIn vertebrate limb morphogenesis, wingless-related integration site (Wnt) proteins and fibroblast growth factors (Fgfs) secreted from the apical ectodermal ridge (AER) coordinate proximodistal outgrowth. Fgfs also sustain sonic hedgehog (Shh) in the zone of polarizing activity (ZPA). Shh directs anteroposterior patterning and expansion and regulates AER-Fgfs, establishing a positive regulatory feedback loop that is vital in sustaining limb outgrowth. The transcription factor LIM homeodomain 2 (Lhx2) is expressed in the distal mesoderm and coordinates AER and ZPA signals that control cellular proliferation, differentiation, and shaping of the developing limb. Yet how Lhx2 is transcriptionally regulated to support such functions has only been partially characterized.Methods/ResultsWe have identified two limb-specific cis-regulatory modules (CRMs) active within the Lhx2 expression domain in the limb. Chromatin conformation analysis of the Lhx2 locus in mouse embryonic limb bud cells predicted CRMs-Lhx2 promoter interactions. Single-cell RNA-sequencing analysis of limb bud cells revealed co-expression of several Fgf-related Ets and Wnt-related Tcf/Lef transcripts in Lhx2-expressing cells. Additionally, disruption of Ets and Tcf/Lef binding sites resulted in loss of reporter-driven CRM activity. Finally, binding of β-catenin to both Lhx2-associated CRMs supports the associated binding of Tcf/Lef transcription factors.DiscussionThese results suggest a role for Ets and Tcf/Lef transcription factors in the regulation of Lhx2 expression through these limb-specific Lhx2-associated CRMs. Moreover, these CRMs provide a mechanism for Fgf and Wnt signaling to localize and maintain distal Lhx2 expression during vertebrate limb development.https://www.frontiersin.org/articles/10.3389/fcell.2025.1552716/fullLhx2vertebrate limb developmentcis-regulatory modulegene regulationFGFsWnt |
| spellingShingle | Jessica C. Britton Anett Somogyi-Leatigaga Billy A. Watson Endika Haro Cassidy G. Mulder Kari D. Kennedy Allen M. Cooper Kristen L. Whitley Ruth-Love Yeboah Jeanyoung Kim Micah C. Yu Micah C. Yu Jairo D. Campos Jairo D. Campos Japhet Amoah Japhet Amoah Shimako Kawauchi Eunyoung Kim Charmaine U. Pira Kerby C. Oberg Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules Frontiers in Cell and Developmental Biology Lhx2 vertebrate limb development cis-regulatory module gene regulation FGFs Wnt |
| title | Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules |
| title_full | Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules |
| title_fullStr | Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules |
| title_full_unstemmed | Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules |
| title_short | Evidence for Fgf and Wnt regulation of Lhx2 during limb development via two limb-specific Lhx2-associated cis-regulatory modules |
| title_sort | evidence for fgf and wnt regulation of lhx2 during limb development via two limb specific lhx2 associated cis regulatory modules |
| topic | Lhx2 vertebrate limb development cis-regulatory module gene regulation FGFs Wnt |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2025.1552716/full |
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