The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.

The equine hoof inner epithelium is folded into primary and secondary epidermal lamellae which increase the dermo-epidermal junction surface area of the hoof and can be affected by laminitis, a common disease of equids. Two keratin proteins (K), K42 and K124, are the most abundant keratins in the ho...

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Main Authors: Caitlin Armstrong, Lynne Cassimeris, Claire Da Silva Santos, Yagmur Micoogullari, Bettina Wagner, Susanna Babasyan, Samantha Brooks, Hannah Galantino-Homer
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2019-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0219234
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author Caitlin Armstrong
Lynne Cassimeris
Claire Da Silva Santos
Yagmur Micoogullari
Bettina Wagner
Susanna Babasyan
Samantha Brooks
Hannah Galantino-Homer
author_facet Caitlin Armstrong
Lynne Cassimeris
Claire Da Silva Santos
Yagmur Micoogullari
Bettina Wagner
Susanna Babasyan
Samantha Brooks
Hannah Galantino-Homer
author_sort Caitlin Armstrong
collection DOAJ
description The equine hoof inner epithelium is folded into primary and secondary epidermal lamellae which increase the dermo-epidermal junction surface area of the hoof and can be affected by laminitis, a common disease of equids. Two keratin proteins (K), K42 and K124, are the most abundant keratins in the hoof lamellar tissue of Equus caballus. We hypothesize that these keratins are lamellar tissue-specific and could serve as differentiation- and disease-specific markers. Our objective was to characterize the expression of K42 and K124 in equine stratified epithelia and to generate monoclonal antibodies against K42 and K124. By RT-PCR analysis, keratin gene (KRT) KRT42 and KRT124 expression was present in lamellar tissue, but not cornea, haired skin, or hoof coronet. In situ hybridization studies showed that KRT124 localized to the suprabasal and, to a lesser extent, basal cells of the lamellae, was absent from haired skin and hoof coronet, and abruptly transitions from KRT124-negative coronet to KRT124-positive proximal lamellae. A monoclonal antibody generated against full-length recombinant equine K42 detected a lamellar keratin of the appropriate size, but also cross-reacted with other epidermal keratins. Three monoclonal antibodies generated against N- and C-terminal K124 peptides detected a band of the appropriate size in lamellar tissue and did not cross-react with proteins from haired skin, corneal limbus, hoof coronet, tongue, glabrous skin, oral mucosa, or chestnut on immunoblots. K124 localized to lamellar cells by indirect immunofluorescence. This is the first study to demonstrate the localization and expression of a hoof lamellar-specific keratin, K124, and to validate anti-K124 monoclonal antibodies.
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spelling doaj.art-f78cffb109aa43ffbe75078069ff1d832022-12-21T18:32:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-01149e021923410.1371/journal.pone.0219234The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.Caitlin ArmstrongLynne CassimerisClaire Da Silva SantosYagmur MicoogullariBettina WagnerSusanna BabasyanSamantha BrooksHannah Galantino-HomerThe equine hoof inner epithelium is folded into primary and secondary epidermal lamellae which increase the dermo-epidermal junction surface area of the hoof and can be affected by laminitis, a common disease of equids. Two keratin proteins (K), K42 and K124, are the most abundant keratins in the hoof lamellar tissue of Equus caballus. We hypothesize that these keratins are lamellar tissue-specific and could serve as differentiation- and disease-specific markers. Our objective was to characterize the expression of K42 and K124 in equine stratified epithelia and to generate monoclonal antibodies against K42 and K124. By RT-PCR analysis, keratin gene (KRT) KRT42 and KRT124 expression was present in lamellar tissue, but not cornea, haired skin, or hoof coronet. In situ hybridization studies showed that KRT124 localized to the suprabasal and, to a lesser extent, basal cells of the lamellae, was absent from haired skin and hoof coronet, and abruptly transitions from KRT124-negative coronet to KRT124-positive proximal lamellae. A monoclonal antibody generated against full-length recombinant equine K42 detected a lamellar keratin of the appropriate size, but also cross-reacted with other epidermal keratins. Three monoclonal antibodies generated against N- and C-terminal K124 peptides detected a band of the appropriate size in lamellar tissue and did not cross-react with proteins from haired skin, corneal limbus, hoof coronet, tongue, glabrous skin, oral mucosa, or chestnut on immunoblots. K124 localized to lamellar cells by indirect immunofluorescence. This is the first study to demonstrate the localization and expression of a hoof lamellar-specific keratin, K124, and to validate anti-K124 monoclonal antibodies.https://doi.org/10.1371/journal.pone.0219234
spellingShingle Caitlin Armstrong
Lynne Cassimeris
Claire Da Silva Santos
Yagmur Micoogullari
Bettina Wagner
Susanna Babasyan
Samantha Brooks
Hannah Galantino-Homer
The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.
PLoS ONE
title The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.
title_full The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.
title_fullStr The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.
title_full_unstemmed The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.
title_short The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.
title_sort expression of equine keratins k42 and k124 is restricted to the hoof epidermal lamellae of equus caballus
url https://doi.org/10.1371/journal.pone.0219234
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