title: Modulation of semaphorin 3A expression by calcium concentration and histamine in human keratinocytes and fibroblasts.
abstract: BACKGROUND
  Both neurotrophins and chemorepellents are involved in the elongation and sprouting of itch-associated C-fibers in the skin. Nerve growth factor (NGF) and semaphorin 3A (Sema3A) are representatives of these two types of axon-guidance factors, respectively.
  
  
  OBJECTIVE
  We investigated the effects of calcium concentration and histamine on the expression of NGF and Sema3A in normal human epidermal keratinocytes (NHEK) and normal human fibroblasts (NHFb).
  
  
  METHODS
  NHEK and NHFb were cultured under different calcium concentrations (0.15-0.9 mM) with or without histamine, and the expression of mRNA for NGF and SEMA3A was assessed by real-time PCR analysis. An immunohistochemical study was performed for Sema3A using normal skin and skin cancer specimens.
  
  
  RESULTS
  In NHEK, SEMA3A expression was elevated by high calcium concentration and reduced by low calcium condition, while NGF expression was not dependent on calcium. Their expressions were unchanged by calcium in NHFb. Immunohistochemically, keratinocytes in the prickle layer of normal epidermis and squamous cell carcinoma cells were positive for Sema3A, sparing basal cells and suprabasal cells. The addition of histamine to NHEK at 10 μg/ml enhanced SEMA3A expression but depressed NGF expression. In NHFb, however, histamine decreased both NGF and SEMA3A levels.
  
  
  CONCLUSIONS
  Sema3A inhibits C-fiber elongation/sprouting in the upper layers of the epidermis, where calcium concentration is high, thereby determining the nerve endings. Histamine reduces Sema3A production by fibroblasts, allowing C-fibers to elongate in the dermis. In contrast, the histamine-augmented keratinocyte production of Sema3A might suppress C-fiber elongation and exaggerated pruritus.
authors.affiliation: Department of Dermatology, University of Occupational and Environmental Health, Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan.
authors.forename: Shoko
authors.initials: S
authors.lastname: Fukamachi
authors.forename: Toshinori
authors.initials: T
authors.lastname: Bito
authors.forename: Noriko
authors.initials: N
authors.lastname: Shiraishi
authors.forename: Miwa
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Kenji
authors.initials: K
authors.lastname: Kabashima
authors.forename: Motonobu
authors.initials: M
authors.lastname: Nakamura
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Tokura
mesh_terms: D002118:Calcium / Q000494:pharmacology; D002280:Basal Cell Carcinoma / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism* / Q000502:physiology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism* / Q000502:physiology; D036421:Nerve Fibers, Unmyelinated / Q000187:drug effects / Q000648:ultrastructure*; D020932:Nerve Growth Factor / Q000187:drug effects / Q000378:metabolism*; D009842:Oligopeptides / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D040121:Semaphorin-3A / Q000187:drug effects / Q000378:metabolism*; D012867:Skin / Q000378:metabolism
chemical_list: D016209:Interleukin-8; D009842:Oligopeptides; D012333:RNA, Messenger; D040121:Semaphorin-3A; C112356:seryl-leucyl-isoleucyl-glycyl--arginyl-leucinamide; D006632:Histamine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D020932:Nerve Growth Factor; D002118:Calcium