Identification of Pax6-dependent gene regulatory networks in the mouse lens

Louise V. Wolf, Ying Yang, Jinhua Wang, Qing Xie, Barbara Braunger, Ernst R. Tamm, Jiri Zavadil, Ales Cvekl

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Lineage-specific DNA-binding transcription factors regulate development by activating and repressing particular set of genes required for the acquisition of a specific cell type. Pax6 is a paired domain and homeodomain-containing transcription factor essential for development of central nervous, olfactory and visual systems, as well as endocrine pancreas. Haploinsufficiency of Pax6 results in perturbed lens development and homeostasis. Loss-of-function of Pax6 is incompatible with lens lineage formation and results in abnormal telencephalic development. Using DNA microarrays, we have identified 559 genes expressed differentially between 1-day old mouse Pax6 heterozygous and wild type lenses. Of these, 178 (31.8%) were similarly increased and decreased in Pax6 homozygous embryonic telencephalon [Holm PC, Mader MT, Haubst N, Wizenmann A, Sigvardsson M, Götz M (2007) Loss- and gain-of-function analyses reveals targets of Pax6 in the developing mouse telencephalon. Mol Cell Neurosci 34: 99-119]. In contrast, 381 (68.2%) genes were differently regulated between the lens and embryonic telencephalon. Differential expression of nine genes implicated in lens development and homeostasis: Cspg2, Igfbp5, Mab21l2, Nrf2f, Olfm3, Spag5, Spock1, Spon1 and Tgfb2, was confirmed by quantitative RT-PCR, with five of these genes: Cspg2, Mab21l2, Olfm3, Spag5 and Tgfb2, identified as candidate direct Pax6 target genes by quantitative chromatin immunoprecipitation (qChIP). In Mab21l2 and Tgfb2 promoter regions, twelve putative individual Pax6-binding sites were tested by electrophoretic mobility shift assays (EMSAs) with recombinant Pax6 proteins. This led to the identification of two and three sites in the respective Mab21l2 and Tgfb2 promoter regions identified by qChIPs. Collectively, the present studies represent an integrative genome-wide approach to identify downstream networks controlled by Pax6 that control mouse lens and forebrain development.

Original languageEnglish (US)
Article numbere4159
JournalPLoS One
Volume4
Issue number1
DOIs
StatePublished - Jan 9 2009

Fingerprint

Gene Regulatory Networks
Lens
Lenses
Telencephalon
Genes
mice
genes
Genetic Promoter Regions
homeostasis
Homeostasis
Transcription Factors
transcription factors
promoter regions
Haploinsufficiency
Chromatin Immunoprecipitation
abnormal development
Recombinant proteins
Electrophoretic Mobility Shift Assay
Electrophoretic mobility
Prosencephalon

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Wolf, L. V., Yang, Y., Wang, J., Xie, Q., Braunger, B., Tamm, E. R., ... Cvekl, A. (2009). Identification of Pax6-dependent gene regulatory networks in the mouse lens. PLoS One, 4(1), [e4159]. https://doi.org/10.1371/journal.pone.0004159

Identification of Pax6-dependent gene regulatory networks in the mouse lens. / Wolf, Louise V.; Yang, Ying; Wang, Jinhua; Xie, Qing; Braunger, Barbara; Tamm, Ernst R.; Zavadil, Jiri; Cvekl, Ales.

In: PLoS One, Vol. 4, No. 1, e4159, 09.01.2009.

Research output: Contribution to journalArticle

Wolf, LV, Yang, Y, Wang, J, Xie, Q, Braunger, B, Tamm, ER, Zavadil, J & Cvekl, A 2009, 'Identification of Pax6-dependent gene regulatory networks in the mouse lens', PLoS One, vol. 4, no. 1, e4159. https://doi.org/10.1371/journal.pone.0004159
Wolf LV, Yang Y, Wang J, Xie Q, Braunger B, Tamm ER et al. Identification of Pax6-dependent gene regulatory networks in the mouse lens. PLoS One. 2009 Jan 9;4(1). e4159. https://doi.org/10.1371/journal.pone.0004159
Wolf, Louise V. ; Yang, Ying ; Wang, Jinhua ; Xie, Qing ; Braunger, Barbara ; Tamm, Ernst R. ; Zavadil, Jiri ; Cvekl, Ales. / Identification of Pax6-dependent gene regulatory networks in the mouse lens. In: PLoS One. 2009 ; Vol. 4, No. 1.
@article{8b447ba364994863b9dd9b3901b295bb,
title = "Identification of Pax6-dependent gene regulatory networks in the mouse lens",
abstract = "Lineage-specific DNA-binding transcription factors regulate development by activating and repressing particular set of genes required for the acquisition of a specific cell type. Pax6 is a paired domain and homeodomain-containing transcription factor essential for development of central nervous, olfactory and visual systems, as well as endocrine pancreas. Haploinsufficiency of Pax6 results in perturbed lens development and homeostasis. Loss-of-function of Pax6 is incompatible with lens lineage formation and results in abnormal telencephalic development. Using DNA microarrays, we have identified 559 genes expressed differentially between 1-day old mouse Pax6 heterozygous and wild type lenses. Of these, 178 (31.8{\%}) were similarly increased and decreased in Pax6 homozygous embryonic telencephalon [Holm PC, Mader MT, Haubst N, Wizenmann A, Sigvardsson M, G{\"o}tz M (2007) Loss- and gain-of-function analyses reveals targets of Pax6 in the developing mouse telencephalon. Mol Cell Neurosci 34: 99-119]. In contrast, 381 (68.2{\%}) genes were differently regulated between the lens and embryonic telencephalon. Differential expression of nine genes implicated in lens development and homeostasis: Cspg2, Igfbp5, Mab21l2, Nrf2f, Olfm3, Spag5, Spock1, Spon1 and Tgfb2, was confirmed by quantitative RT-PCR, with five of these genes: Cspg2, Mab21l2, Olfm3, Spag5 and Tgfb2, identified as candidate direct Pax6 target genes by quantitative chromatin immunoprecipitation (qChIP). In Mab21l2 and Tgfb2 promoter regions, twelve putative individual Pax6-binding sites were tested by electrophoretic mobility shift assays (EMSAs) with recombinant Pax6 proteins. This led to the identification of two and three sites in the respective Mab21l2 and Tgfb2 promoter regions identified by qChIPs. Collectively, the present studies represent an integrative genome-wide approach to identify downstream networks controlled by Pax6 that control mouse lens and forebrain development.",
author = "Wolf, {Louise V.} and Ying Yang and Jinhua Wang and Qing Xie and Barbara Braunger and Tamm, {Ernst R.} and Jiri Zavadil and Ales Cvekl",
year = "2009",
month = "1",
day = "9",
doi = "10.1371/journal.pone.0004159",
language = "English (US)",
volume = "4",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "1",

}

TY - JOUR

T1 - Identification of Pax6-dependent gene regulatory networks in the mouse lens

AU - Wolf, Louise V.

AU - Yang, Ying

AU - Wang, Jinhua

AU - Xie, Qing

AU - Braunger, Barbara

AU - Tamm, Ernst R.

AU - Zavadil, Jiri

AU - Cvekl, Ales

PY - 2009/1/9

Y1 - 2009/1/9

N2 - Lineage-specific DNA-binding transcription factors regulate development by activating and repressing particular set of genes required for the acquisition of a specific cell type. Pax6 is a paired domain and homeodomain-containing transcription factor essential for development of central nervous, olfactory and visual systems, as well as endocrine pancreas. Haploinsufficiency of Pax6 results in perturbed lens development and homeostasis. Loss-of-function of Pax6 is incompatible with lens lineage formation and results in abnormal telencephalic development. Using DNA microarrays, we have identified 559 genes expressed differentially between 1-day old mouse Pax6 heterozygous and wild type lenses. Of these, 178 (31.8%) were similarly increased and decreased in Pax6 homozygous embryonic telencephalon [Holm PC, Mader MT, Haubst N, Wizenmann A, Sigvardsson M, Götz M (2007) Loss- and gain-of-function analyses reveals targets of Pax6 in the developing mouse telencephalon. Mol Cell Neurosci 34: 99-119]. In contrast, 381 (68.2%) genes were differently regulated between the lens and embryonic telencephalon. Differential expression of nine genes implicated in lens development and homeostasis: Cspg2, Igfbp5, Mab21l2, Nrf2f, Olfm3, Spag5, Spock1, Spon1 and Tgfb2, was confirmed by quantitative RT-PCR, with five of these genes: Cspg2, Mab21l2, Olfm3, Spag5 and Tgfb2, identified as candidate direct Pax6 target genes by quantitative chromatin immunoprecipitation (qChIP). In Mab21l2 and Tgfb2 promoter regions, twelve putative individual Pax6-binding sites were tested by electrophoretic mobility shift assays (EMSAs) with recombinant Pax6 proteins. This led to the identification of two and three sites in the respective Mab21l2 and Tgfb2 promoter regions identified by qChIPs. Collectively, the present studies represent an integrative genome-wide approach to identify downstream networks controlled by Pax6 that control mouse lens and forebrain development.

AB - Lineage-specific DNA-binding transcription factors regulate development by activating and repressing particular set of genes required for the acquisition of a specific cell type. Pax6 is a paired domain and homeodomain-containing transcription factor essential for development of central nervous, olfactory and visual systems, as well as endocrine pancreas. Haploinsufficiency of Pax6 results in perturbed lens development and homeostasis. Loss-of-function of Pax6 is incompatible with lens lineage formation and results in abnormal telencephalic development. Using DNA microarrays, we have identified 559 genes expressed differentially between 1-day old mouse Pax6 heterozygous and wild type lenses. Of these, 178 (31.8%) were similarly increased and decreased in Pax6 homozygous embryonic telencephalon [Holm PC, Mader MT, Haubst N, Wizenmann A, Sigvardsson M, Götz M (2007) Loss- and gain-of-function analyses reveals targets of Pax6 in the developing mouse telencephalon. Mol Cell Neurosci 34: 99-119]. In contrast, 381 (68.2%) genes were differently regulated between the lens and embryonic telencephalon. Differential expression of nine genes implicated in lens development and homeostasis: Cspg2, Igfbp5, Mab21l2, Nrf2f, Olfm3, Spag5, Spock1, Spon1 and Tgfb2, was confirmed by quantitative RT-PCR, with five of these genes: Cspg2, Mab21l2, Olfm3, Spag5 and Tgfb2, identified as candidate direct Pax6 target genes by quantitative chromatin immunoprecipitation (qChIP). In Mab21l2 and Tgfb2 promoter regions, twelve putative individual Pax6-binding sites were tested by electrophoretic mobility shift assays (EMSAs) with recombinant Pax6 proteins. This led to the identification of two and three sites in the respective Mab21l2 and Tgfb2 promoter regions identified by qChIPs. Collectively, the present studies represent an integrative genome-wide approach to identify downstream networks controlled by Pax6 that control mouse lens and forebrain development.

UR - http://www.scopus.com/inward/record.url?scp=58349090476&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58349090476&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0004159

DO - 10.1371/journal.pone.0004159

M3 - Article

C2 - 19132093

AN - SCOPUS:58349090476

VL - 4

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 1

M1 - e4159

ER -