title: Mechanosensory and ATP Release Deficits following Keratin14-Cre-Mediated TRPA1 Deletion Despite Absence of TRPA1 in Murine Keratinocytes.
abstract: Keratinocytes are the first cells that come into direct contact with external tactile stimuli; however, their role in touch transduction in vivo is not clear. The ion channel Transient Receptor Potential Ankyrin 1 (TRPA1) is essential for some mechanically-gated currents in sensory neurons, amplifies mechanical responses after inflammation, and has been reported to be expressed in human and mouse skin. Other reports have not detected Trpa1 mRNA transcripts in human or mouse epidermis. Therefore, we set out to determine whether selective deletion of Trpa1 from keratinocytes would impact mechanosensation. We generated K14Cre-Trpa1fl/fl mice lacking TRPA1 in K14-expressing cells, including keratinocytes. Surprisingly, Trpa1 transcripts were very poorly detected in epidermis of these mice or in controls, and detection was minimal enough to preclude observation of Trpa1 mRNA knockdown in the K14Cre-Trpa1fl/fl mice. Unexpectedly, these K14Cre-Trpa1fl/fl mice nonetheless exhibited a pronounced deficit in mechanosensitivity at the behavioral and primary afferent levels, and decreased mechanically-evoked ATP release from skin. Overall, while these data suggest that the intended targeted deletion of Trpa1 from keratin 14-expressing cells of the epidermis induces functional deficits in mechanotransduction and ATP release, these deficits are in fact likely due to factors other than reduction of Trpa1 expression in adult mouse keratinocytes because they express very little, if any, Trpa1.
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Zappia
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Sheldon R
authors.initials: SR
authors.lastname: Garrison
authors.affiliation: Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Oleg
authors.initials: O
authors.lastname: Palygin
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Andy D
authors.initials: AD
authors.lastname: Weyer
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Marie E
authors.initials: ME
authors.lastname: Barabas
authors.affiliation: Division of Pediatric Pathology, Department of Pathology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Lawlor
authors.affiliation: Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Alexander
authors.initials: A
authors.lastname: Staruschenko
authors.affiliation: Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Cheryl L
authors.initials: CL
authors.lastname: Stucky
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000344:Afferent Pathways / Q000502:physiology; D000818:Animals; D020296:Animals, Congenic; D001169:Arthritis, Experimental / Q000503:physiopathology; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D005620:Freund's Adjuvant / Q000633:toxicity; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D017930:Genes, Reporter; D019426:Integrases; D015603:Keratinocytes / Q000378:metabolism*; D008465:Mechanoreceptors / Q000502:physiology*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D059225:Nociception / Q000502:physiology; D009928:Organ Specificity; D017288:Pain Threshold / Q000502:physiology; D010812:Physical Stimulation / Q000009:adverse effects; D012333:RNA, Messenger / Q000096:biosynthesis; D011984:Sensory Receptor Cells / Q000502:physiology; D012867:Skin / Q000166:cytology / Q000196:embryology; D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels / Q000096:biosynthesis / Q000172:deficiency* / Q000235:genetics / Q000502:physiology
chemical_list: D012333:RNA, Messenger; D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels; C475685:Trpa1 protein, mouse; D000255:Adenosine Triphosphate; D005620:Freund's Adjuvant; C045073:Cre recombinase; D019426:Integrases