QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 15 August 2006
doi: 10.1242/jcs.03070

This Article Summary Full Text Full Text (PDF) Supplementary Material Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sur, I. Articles by Toftgård, R. Search for Related Content PubMed PubMed Citation Articles by Sur, I. Articles by Toftgård, R. Social Bookmarking                         What's this?

Epidermal and craniofacial defects in mice overexpressing Klf5 in the basal layer of the epidermis

¹ Department of Bioscience and Nutrition, Clinical Research Center, and Department of Laboratory Medicine Division of Pathology, Karolinska Institutet, Novum, SE-141 57 Huddinge, Sweden
² Unit for Morphological Phenotype Analysis, Clinical Research Center, and Department of Laboratory Medicine Division of Pathology, Karolinska Institutet, Novum, SE-141 57 Huddinge, Sweden

View larger version (49K): [in a new window]   Fig. 1. Cranioabdominoschisis and ectodermal dysplasia in bi-transgenics. (a) Overall morphology of wild-type and K5tTA/TRE-KLF5 bi-transgenic embryos at E16.5. (b) Skeletons of E17.5 embryos. The skeletons were double-stained with Alizarin Red and Alcian Blue. Bone is stained red and cartilage is blue. (c) Skull sections of E16.5 embryos. Bars, 500 µm. (1) Haematoxylin-Eosin-stained skull sections showing absence of vibrissae follicles and incisors in the bi-transgenic embryo. (2) Immunohistochemical staining with anti-keratin5 antibody showing the arrested molar development at the early bud-stage in the bi-transgenic embryo. (3) Immunohistochemical staining with anti-keratin5 antibody showing the absence of eyelid formation in the bi-transgenic embryo. (d) Expression of the KLF5 transgene in E16.5 embryos analysed by (1) RT-PCR and (2) immunohistochemistry using anti-Klf5 antibody. Bar, 50 µm.

View larger version (60K): [in a new window]   Fig. 2. Hypoproliferative epidermis and skin barrier defects in the bi-transgenics. (a) Decreased proliferation in the epidermis of the K5tTA/TRE-Klf5 bi-transgenic embryos (E16.5) compared with the wild type. Haematoxylin-Eosin-stained sections of dorsal skin at E16.5 (upper panel). Immunohistochemical staining of E16.5 dorsal skin with the anti-Ki67 antibody (lower panel). Bar, 50 µm. (b) Defective skin barrier in a bi-transgenic embryo compared with a wild-type embryo at E17.5.

View larger version (56K): [in a new window]   Fig. 3. Altered expression of cytokeratin markers in the epidermis of the bi-transgenics. Immunohistochemical staining of dorsal skin from E18.5 embryos with anti-keratin5, loricrin, anti-keratin1, anti-keratin17 or anti-keratin8 antibodies showing abnormal expression of keratin1, 8, 17 but normal compartmentalization of keratin5 and loricrin. Bar, 50 µm.

View larger version (82K): [in a new window]   Fig. 4. Abnormal epidermal morphology in the bi-transgenics. Ultrastructural analysis of wild-type and bi-transgenic epidermis from E14.5 embryos. Magnification, x3800. Inset shows higher magnification (x107,000) view of granules (marked by arrow) observed in the suprabasal cells of the bi-transgenic embryos.

View larger version (50K): [in a new window]   Fig. 5. Reduced p63 expression in the bi-transgenics. (a) Immunohistochemical staining of a dorsal skin section from E18.5 wild-type and K5tTA/TRE-KLF5 bi-transgenic embryos with anti-p63 (4A4) antibody. Bar, 50 µm. (b) RT-PCR analysis of different transcripts from skin of E16.5 and E18.5 embryos. Lane 1, E16.5 wild type; lane 2, E16.5 bi-transgenic; lane 3, E18.5 wild type; lane 4, E18.5 bi-transgenic.

View larger version (37K): [in a new window]   Fig. 6. Growth defect of bi-transgenic keratinocytes in vitro. (a) Image of 7-day old keratinocyte cultures from wild-type and K5tTA/TRE-KLF5 pups. Bar, 50 µm. (b) Growth curves of wild-type () and bi-transgenic keratinocytes () determined using the WST-1 reagent. The data are representative of three bi-transgenics and three wild-type cultures. (c) Addition of doxycyline to the culture prevents the growth defect; wild type (blue bars), bi-transgenic (pink bars). (d) Immunofluorescent detection and quantification of BrdU incorporation in wild-type (blue bars) and bi-transgenic (pink bars) keratinocytes showing a proliferation block in the bi-transgenics. Bar, 50 µm. In c-d the numbers represent the mean and error bars ±s.e.m. from two bi-transgenic and two wild-type cultures. To quantify BrdU incorporation at least 500 cells were counted for each culture in five fields.

View larger version (47K): [in a new window]   Fig. 7. Overexpression of Klf5 in the adult epidermis. (a) Hyperkeratosis, follicle occlusion and hyperproliferation after induction of KLF5-transgene in the bi-transgenic adult mice. Hyperproliferation of keratinocytes is mainly observed at the edges of epidermal erosions. Bar, 50 µm. (b) Analysis of expression of 6 integrin and CD34 markers in keratinocytes by flow cytometry. Keratinocytes were isolated from normal and bi-transgenic adult female mice in which Klf5 overexpression was initiated at 4-5 weeks of age. Keratinocytes were stained with CD34-FITC and 6 integrin-PE-Cy5 and examined by flow cytometry. The bi-transgenics 1 and 2 were 3 months old and the bi-transgenic 3 female and the normal control were 5 months old at the time of analysis.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

Twitter