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Epithelial membrane pumps, channels and transporters as phenotypic markers of equine hoof keratinocytes in situ : an immunohistochemical study
Author(s) -
Asplin Katie,
Harris Pat,
Mobasheri Ali
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.1002.23
Subject(s) - hoof , biology , microbiology and biotechnology , epithelial sodium channel , phenotype , epithelium , chemistry , anatomy , biochemistry , sodium , genetics , organic chemistry , gene
Several different enzymatic techniques have been published for isolating and culturing primary equine hoof keratinocytes. However, the phenotype of the resulting primary cultures is poorly understood. Freshly isolated hoof cultures consist of a mixture of keratinocytes and fibroblasts, with the latter outgrowing and overtaking the keratinocytes. Although monoclonal antibodies against cytokeratins and vimentin have been used in recent studies to investigate the differentiation of hoof epidermal cells, there has been little progress on the basic cell biology of equine hoof keratinocytes and the markers that they express in vivo . In this study we hypothesized that epithelial transport proteins may be used as phenotypic markers for these cells. To test this hypothesis we employed a panel of monoclonal and polyclonal antibodies against a range of epithelial ion and solute transporters to establish new phenotypic markers for these cells in situ . Equine hoof keratinocytes showed positive and preferential immunoreactivity for several transporters and ion channels compared to other cell types in the hoof. These included Na, K‐ATPase, Na, K, 2Cl co‐transporter, epithelial sodium channels (ENaC), Maxi K (BK) calcium‐activated potassium channels, and aquaporins including AQP3 and AQP9. These results suggest that epithelial membrane transporters are specific and sensitive “metabolic” markers of hoof keratinocytes.