title: Ultrastructural localization of histidine-labelled interkeratin matrix during keratinization of amphibian epidermis.
abstract: Interkeratin histidine-rich proteins (filaggrins) play a functional role in aggregation of keratin filaments into dense bundles during terminal differentiation of mammalian keratinocytes when forming the dense matrix of the stratum corneum. The origin of the stratum corneum during adaptation to land in amphibious vertebrate progenitors was probably linked with the synthesis of matrix proteins. However, whether similar proteins are present in living amphibians is unknown. The possible involvement of interkeratin matrix molecules rich in basic amino acids such as histidine during keratinization of amphibian epidermis has been evaluated in the present study by ultrastructural autoradiography after administration of tritiated histidine. At 4 and 8 h post-injection, labelling was mainly localized over electron-dense amorphous material or irregular granules in between keratin filaments in cells of the upper intermedium, replacement, and immature corneous layers. Nuclear material incorporating tritiated histidine was also present in the maturing corneous layer. Small mucous-like granules did not take up tritiated histidine and X-ray microanalysis indicated that the latter granules contained sulphur. The present study suggests that small amounts of histidine-rich molecules which were not sufficient to form microscopically-visible keratohyalin granules were present in ancestral amphibian epidermis. However, this material was sufficient to promote aggregation of keratin filaments in the cytoplasm of amphibian differentiating keratinocytes, especially near the external corneous cell envelope. Electron-dense material associated with the corneous cell envelope also contained sulphur as indicated by X-ray microanalysis. It is unknown whether sulphur is derived from either sulphated mucins, or disulphide bonds in aggregated keratins, or specific sulphur-rich proteins.
mesh_terms: D000818:Animals; D001345:Autoradiography; D002025:Bufonidae / Q000033:anatomy & histology*; D004817:Epidermis / Q000737:chemistry* / Q000648:ultrastructure*; D000091344:Filaggrin Proteins; D006639:Histidine / Q000032:analysis; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007382:Intermediate Filaments / Q000737:chemistry / Q000648:ultrastructure; D007633:Keratins / Q000032:analysis / Q000648:ultrastructure*; D011506:Proteins / Q000032:analysis*; D014316:Tritium / Q000032:analysis; D000099053:Histidine-Rich Glycoprotein
chemical_list: D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D011506:Proteins; D000099053:Histidine-Rich Glycoprotein; D014316:Tritium; D006639:Histidine; D007633:Keratins