A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium

Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corne...
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Autor*in:	Natalie J. Dorà [verfasserIn] Martine Manuel [verfasserIn] Dirk-Jan Kleinjan [verfasserIn] David J. Price [verfasserIn] J. Martin Collinson [verfasserIn] Robert E. Hill [verfasserIn] John D. West [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2018

Schlagwörter:	Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6

Übergeordnetes Werk:	In: BMC Research Notes - BMC, 2008, 11(2018), 1, Seite 8
Übergeordnetes Werk:	volume:11 ; year:2018 ; number:1 ; pages:8

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/s13104-018-3812-9

Katalog-ID:	DOAJ046987789

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520			\|a Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs.
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allfields	10.1186/s13104-018-3812-9 doi (DE-627)DOAJ046987789 (DE-599)DOAJb408ec98da0b4083aacabda28374eb4e DE-627 ger DE-627 rakwb eng QH301-705.5 Q1-390 Natalie J. Dorà verfasserin aut A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs. Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6 Medicine R Biology (General) Science (General) Martine Manuel verfasserin aut Dirk-Jan Kleinjan verfasserin aut David J. Price verfasserin aut J. Martin Collinson verfasserin aut Robert E. Hill verfasserin aut John D. West verfasserin aut In BMC Research Notes BMC, 2008 11(2018), 1, Seite 8 (DE-627)559431805 (DE-600)2413336-X 17560500 nnns volume:11 year:2018 number:1 pages:8 https://doi.org/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/article/b408ec98da0b4083aacabda28374eb4e kostenfrei http://link.springer.com/article/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/toc/1756-0500 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2018 1 8
spelling	10.1186/s13104-018-3812-9 doi (DE-627)DOAJ046987789 (DE-599)DOAJb408ec98da0b4083aacabda28374eb4e DE-627 ger DE-627 rakwb eng QH301-705.5 Q1-390 Natalie J. Dorà verfasserin aut A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs. Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6 Medicine R Biology (General) Science (General) Martine Manuel verfasserin aut Dirk-Jan Kleinjan verfasserin aut David J. Price verfasserin aut J. Martin Collinson verfasserin aut Robert E. Hill verfasserin aut John D. West verfasserin aut In BMC Research Notes BMC, 2008 11(2018), 1, Seite 8 (DE-627)559431805 (DE-600)2413336-X 17560500 nnns volume:11 year:2018 number:1 pages:8 https://doi.org/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/article/b408ec98da0b4083aacabda28374eb4e kostenfrei http://link.springer.com/article/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/toc/1756-0500 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2018 1 8
allfields_unstemmed	10.1186/s13104-018-3812-9 doi (DE-627)DOAJ046987789 (DE-599)DOAJb408ec98da0b4083aacabda28374eb4e DE-627 ger DE-627 rakwb eng QH301-705.5 Q1-390 Natalie J. Dorà verfasserin aut A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs. Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6 Medicine R Biology (General) Science (General) Martine Manuel verfasserin aut Dirk-Jan Kleinjan verfasserin aut David J. Price verfasserin aut J. Martin Collinson verfasserin aut Robert E. Hill verfasserin aut John D. West verfasserin aut In BMC Research Notes BMC, 2008 11(2018), 1, Seite 8 (DE-627)559431805 (DE-600)2413336-X 17560500 nnns volume:11 year:2018 number:1 pages:8 https://doi.org/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/article/b408ec98da0b4083aacabda28374eb4e kostenfrei http://link.springer.com/article/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/toc/1756-0500 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2018 1 8
allfieldsGer	10.1186/s13104-018-3812-9 doi (DE-627)DOAJ046987789 (DE-599)DOAJb408ec98da0b4083aacabda28374eb4e DE-627 ger DE-627 rakwb eng QH301-705.5 Q1-390 Natalie J. Dorà verfasserin aut A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs. Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6 Medicine R Biology (General) Science (General) Martine Manuel verfasserin aut Dirk-Jan Kleinjan verfasserin aut David J. Price verfasserin aut J. Martin Collinson verfasserin aut Robert E. Hill verfasserin aut John D. West verfasserin aut In BMC Research Notes BMC, 2008 11(2018), 1, Seite 8 (DE-627)559431805 (DE-600)2413336-X 17560500 nnns volume:11 year:2018 number:1 pages:8 https://doi.org/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/article/b408ec98da0b4083aacabda28374eb4e kostenfrei http://link.springer.com/article/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/toc/1756-0500 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2018 1 8
allfieldsSound	10.1186/s13104-018-3812-9 doi (DE-627)DOAJ046987789 (DE-599)DOAJb408ec98da0b4083aacabda28374eb4e DE-627 ger DE-627 rakwb eng QH301-705.5 Q1-390 Natalie J. Dorà verfasserin aut A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs. Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6 Medicine R Biology (General) Science (General) Martine Manuel verfasserin aut Dirk-Jan Kleinjan verfasserin aut David J. Price verfasserin aut J. Martin Collinson verfasserin aut Robert E. Hill verfasserin aut John D. West verfasserin aut In BMC Research Notes BMC, 2008 11(2018), 1, Seite 8 (DE-627)559431805 (DE-600)2413336-X 17560500 nnns volume:11 year:2018 number:1 pages:8 https://doi.org/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/article/b408ec98da0b4083aacabda28374eb4e kostenfrei http://link.springer.com/article/10.1186/s13104-018-3812-9 kostenfrei https://doaj.org/toc/1756-0500 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2018 1 8
language	English
source	In BMC Research Notes 11(2018), 1, Seite 8 volume:11 year:2018 number:1 pages:8
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callnumber-first	Q - Science
author	Natalie J. Dorà
spellingShingle	Natalie J. Dorà misc QH301-705.5 misc Q1-390 misc Cornea misc Corneal epithelium misc Cre-loxP misc CAG-CreER misc Krt19-CreER misc Pax6 misc Medicine misc R misc Biology (General) misc Science (General) A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium
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topic_title	QH301-705.5 Q1-390 A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium Cornea Corneal epithelium Cre-loxP CAG-CreER Krt19-CreER Pax6
topic	misc QH301-705.5 misc Q1-390 misc Cornea misc Corneal epithelium misc Cre-loxP misc CAG-CreER misc Krt19-CreER misc Pax6 misc Medicine misc R misc Biology (General) misc Science (General)
topic_unstemmed	misc QH301-705.5 misc Q1-390 misc Cornea misc Corneal epithelium misc Cre-loxP misc CAG-CreER misc Krt19-CreER misc Pax6 misc Medicine misc R misc Biology (General) misc Science (General)
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title	A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium
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title_full	A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium
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title_sort	conditional pax6 depletion study with no morphological effect on the adult mouse corneal epithelium
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title_auth	A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium
abstract	Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs.
abstractGer	Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs.
abstract_unstemmed	Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs.
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title_short	A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium
url	https://doi.org/10.1186/s13104-018-3812-9 https://doaj.org/article/b408ec98da0b4083aacabda28374eb4e http://link.springer.com/article/10.1186/s13104-018-3812-9 https://doaj.org/toc/1756-0500
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Dorà</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="2"><subfield code="a">A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2018</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">Text</subfield><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">Computermedien</subfield><subfield code="b">c</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield><subfield code="b">cr</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Abstract Objective The corneas of heterozygous Pax6 +/− mice develop abnormally and deteriorate further after birth but it is not known whether the postnatal deterioration is predetermined by abnormal development. Our objective was to identify whether depletion of Pax6 in adult mice caused any corneal abnormalities, similar to those in Pax6 +/− mice, where Pax6 levels are low throughout development and adulthood. We used two tamoxifen-inducible, Cre-loxP experimental strategies to deplete Pax6 either ubiquitously or in a restricted range of cell types. Results In a preliminary study, ubiquitous depletion of Pax6 by tamoxifen treatment of E9.5 CAG-CreER Tg/−;Pax6 fl/fl embryos affected eye development. Tamoxifen treatment of 12-week old, adult CAG-CreER Tg/−;Pax6 fl/+ and CAG-CreER Tg/−;Pax6 fl/fl mice resulted in weak and/or patchy Pax6 immunostaining in the corneal epithelium but caused no corneal abnormalities. GFP staining in tamoxifen-treated CAG-CreER Tg/−;RCE:loxP reporter mice was also patchy. We attribute patchy Pax6 staining to mosaic deletion of the Pax6 fl allele, probably caused by mosaic CAG-CreER Tg expression. In a parallel study, we treated adult Krt19-CreER Tg/−;Pax6 fl/+ mice with tamoxifen to try to deplete Pax6 in limbal epithelial stem cells (LESCs) which replenish the corneal epithelium. However, Pax6 staining remained strong after a 12-week chase period so the Krt19-CreER Tg/− transgene may have failed to target LESCs.</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Cornea</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Corneal epithelium</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Cre-loxP</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">CAG-CreER</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Krt19-CreER</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Pax6</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">Medicine</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">R</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">Biology (General)</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">Science (General)</subfield></datafield><datafield tag="700" ind1="0" ind2=" "><subfield code="a">Martine Manuel</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="700" ind1="0" ind2=" "><subfield code="a">Dirk-Jan Kleinjan</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="700" ind1="0" ind2=" "><subfield code="a">David J. 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A conditional Pax6 depletion study with no morphological effect on the adult mouse corneal epithelium

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