title: Formation of cornified cell envelope in human hair follicle development.
abstract: BACKGROUND
  Cornified cell envelope (CCE) formation is an important step in the final stage of keratinization, in which CCE precursor proteins including involucrin and loricrin are cross-linked by keratinocyte transglutaminases (TGases) to the inner surface of the plasma membrane of cornified cells, while the outer surface is coated with material derived from secreted lamellar granules.
  
  
  OBJECTIVE
  Skin samples from human fetuses of a series of estimated gestational age (EGA) (49-163 days) were studied for the prescence of precursor proteins. Methods TGase activity was studied by in situ TGase activity assay, and ultrastructural features of CCE formation were observed at each stage of hair follicle development. We used immunofluorescent labelling to investigate the time and site of expression of CCE precursor proteins involucrin and loricrin, TGases 1, 2 and 3, and a 25-kDa lamellar granule-associated protein (LGP) in developing human hair follicles.
  
  
  RESULTS
  In the hair germ (65-84 days EGA) (corresponding to the stages 1-2 of murine hair follicle morphogenesis), only TGase 2 was observed in the entire hair germ, where in situ TGase activity was weakly positive, although thickening of cell membrane was not seen ultrastructurally. In the hair peg (85-104 days EGA) (corresponding to the stage 3 of murine hair follicle morphogenesis), loricrin and TGase 2 were seen in cells of the upper part of the hair peg while TGase 1, 3 and LGP were observed in the inner cells of the hair peg. In situ TGase activity was weakly positive in the upper part and inner cells of the hair peg. In the bulbous hair peg (105-135 days EGA) (corresponding to the stages 4-6 of murine hair follicle morphogenesis) and differentiated lanugo hair follicle (> 135 days EGA) (corresponding to the stages 7-8 of murine hair follicle morphogenesis), immunoreactivities of involucrin, loricrin, TGase 1, 2, 3, in situ TGase activity and LGP were detected in the inner root sheath cells, hair canals and inner cells of the outer root sheath in the region of the isthmus. Ultrastructurally, thickening of cell membrane was already seen in the inner root sheath cells of the bulbous hair peg and electron-dense, thick CCE was observed in the hair cuticle and hair canal of differentiated lanugo hair follicle.
  
  
  CONCLUSIONS
  These data indicate that, in terms of CCE formation, certain portions of the developing human hair follicle have already been determined in differentiation of the hair canal and cuticle at the hair peg stage.
authors.affiliation: Department of Dermatology, Hokkaido University Graduate School of Medicine, North 15 West 7, Kita-ku, Sapporo 060-8638, Japan. akiyama@med.hokudai.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Akiyama
authors.forename: I
authors.initials: I
authors.lastname: Matsuo
authors.forename: H
authors.initials: H
authors.lastname: Shimizu
mesh_terms: D002454:Cell Differentiation; D005455:Fluorescent Antibody Technique; D018859:Hair Follicle / Q000737:chemistry / Q000196:embryology*; D006801:Humans; D015603:Keratinocytes / Q000201:enzymology*; D008565:Membrane Proteins / Q000032:analysis; D009024:Morphogenesis / Q000502:physiology; D011498:Protein Precursors / Q000032:analysis; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D011498:Protein Precursors; C064554:loricrin; C032253:involucrin; D011503:Transglutaminases