Immunocytochemical evaluation of reorganisation of keratinocyte cytoskeleton induced by change in Ca2plus concentration in culture medium

• Autorzy: Saczko J , Gebarowska E. , Zabel M.
• Języki publikacji: EN

Abstrakty

EN

Ca²⁺ level-induced changes in the arrangement of cytoskeleton of cultured keratinocytes were estimated immunocytochemically, by evaluating expression of specific cytokeratins, desmoplakin and tubulin. Keratinocytes were isolated from fragments of skin of dead human foetuses. Culture of epidermal cells was performed in two phases: phase I yielded cells of high proliferation abilities in serumfree Keratinocyte SFM of low Ca²⁺ level (0.03 mM); in phase II differentiated cells were obtained in Dulbecco medium of a high Ca²⁺ concentration (1.2 mM). Immunocytochemical evaluation of phase I and II cells revealed an array of differences which involved mainly expression and distribution of specific cytokeratins, distribution of tubulin, testifying to a different microtubule arrangement and distribution of desmoplakin and indicating a tendency to form desmosomes. The changes were induced by the changes in Ca²⁺ level in the culture medium.

Słowa kluczowe

EN

epidermal cell; cytoskeleton; calcium; keratinocyte; proliferation; man; culture medium; immunocytochemistry; cytokeratin; desmosome; fetus

• Wydawca: -
• Czasopismo: Folia Morphologica
• Rocznik: 2000
• Tom: 59
• Numer: 4
• Opis fizyczny: p.247-251,fig.

Twórcy

autor

Saczko J

• Medical University of Wroclaw, Chalubinskiego 10, 50-368 Wroclaw, Poland

autor

Gebarowska E.

autor

Zabel M.

Bibliografia

• 1. Chaturvedi V, Qin JZ, Denning MF, Choubey D, Diaz MO, Nickoloff BJ (1999) Apoptosis in proliferating senescent, immortalized keratinocytes. J Biol Chem, 274: 23358–23367.
• 2. Daley JP, Donovan JM (1994) Growth of human keratinocytes in serum-free medium. Cell Biology: A Laboratory Handbook. Celis JE [Ed], pp 90–95.
• 3. Gandarillas A, Goldsmith LA, Gshmeissner S, Leigh IM, Watt FM (1999) Evidence that apoptosis and terminal differentiation of epidermal keratinocytes are distinct processes. Exp Dermatol, 8: 71–79.
• 4. Girolomani G, Stone DK, Bergstresser PR, Cruz PD (1992) Increased number and microtubule associated dispersal of aciadic intracellular compartments accompany differentiation of cultured human keratinocytes. J Invest Dermatol, 98: 911–917.
• 5. Haftek M., Hansen MU, Kaiser HW, Kreysel HW, Smitt D (1996) Interkeratinocyte adherens junctions: immunocytochemical visualisation of cell-cell junctional structures, distinct from desmosomes, in human epidermis. J Invest Dermatol, 106: 498–504.
• 6. Jensen PKA, Norgard JOR, Knudsen CH, Nielsen V, Bolund L (1990) Effects of extra and intracellular calcium concentration on DNA replication, lateral growth, and differentiation of human epidermal cells in culture. Virchovs Archiv B Cell Pathol, 59: 17–25.
• 7. Jones PPH, Watt FM (1993) Separation of human epidermal stem cells from transit amplifying cells on the basis of differences in integrin function and expression. Cell, 73: 713–724.
• 8. Katagata Y, Aoki T, Kondo S (1999) Detecting expression of keratins 8/18 in human HaCaT keratinocytes. J Dermatol Sci, 19: 139–143.
• 9. Lloyd C, Cheng QCYUJ, Furksen K, Degestein L, Hutton E, Fuchs E (1995) The basal keratin network of stratified squamous epithelia: defining K15 function in the absence of K14. J Cell Biol, 106: 1249–1261.
• 10. Phaire-Washington I, Silverstin S. C, Wang E (1980) Phorbol myristate acetate stimulates microtubule and 10nm filament extension and lisosome redistribution in mouse macrophages J Cell Biol, 86: 641–655.
• 11. Porter RM, Leitgeb S, Melton DW, Swensson O, Eady RAJ, Magin TM (1996) Gene targeting at the mouse cytokeratin 10 locus: severe skin fragility and changes of cytokeratin expression in the epidermis. J Cell Biol, 132: 925–936.
• 12. Shremer EJ, Jester JV, Hardy C, Milano D, T-Sun T (1989) Transient synthesis of K6 and K 16 keratins in regenerating rabbit corneal epithelim: keratin markers for an alternative pathway of keratinocyte differentiation. Differentiation, 42: 103–110.
• 13. Teraci Y, Shiohara T (1999) Apoptosis and the skin. Eur J Dermatol, 9: 413–425.
• 14. Troy TC, Turksen K (1999) In vitro characteristics of early epidermal progenitors isolated from keratin 14 (K14)- deficient mice: insights into the role of keratin 17 in mouse keratinocytes. J Cell Physiol, 180: 409–421.
• 15. Tu CL, Oda Y, Bikle DD (1999) Effects of a calcium receptor activator on the cellular response to calcium in human keratinocytes. J Invest Dermatol, 113: 340–345.
• 16. Wanner R, Wolff B, Głowacki F, Kolde G, Wittia B (1999) The loss of desmosomes after retinoic acid treatment results in an apparent inhibition of HaCaT keratinocyte differentiation. Arch Dermatol Res, 291: 346–353.
• 17. Watt FM, Mattey DL, Carrod DR (1984) Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J Cell Biol, 99: 2211–2215.
• 18. Yuspa SH, Kilkenny AE, Steinert PM, Roop DR (1989) Expression of murine epidermal differentiation markers is tightly regulated by restricted extracellular calcium concentrations in vitro. J Cell Biol, 109: 1207–1217.
• 19. Zamansky GB, Nguyen U Chou IN (1992) An immunofluorescence study of the effects of ultraviolet radiation on the organization of microfilaments, keratin intermediate filaments, and microtubules in human keratinocytes. Cell Motil Cytoskeleton, 22: 296–306.
• 20. Zamansky GB, Nguyen U, Chou IN (1991) An immunofluorescence study of the calcium-induced coordinated reorganization of microfilaments, keratin intermediate filaments and microtubules in cultured human keratinocytes. J Invest Dermatol, 97: 985–994.