Involucrin and SPRR are synthesized sequentially in differentiating cultured epidermal cells

Epidermal keratinocytes form cornified cell envelopes during terminal differentiation. These envelopes are composed of several cross-linked molecules, including involucrin, loricrin, and SPRR. We have previously reported that involucrin is synthesized earlier in terminal differentiation than loricrin. To further elucidate the mechanisms of terminal differentiation, we have now examined the expression of the two differentiation markers, involucrin and SPRR, in cultured human epidermal keratinocytes. In confluent nonstratified cultures, many involucrin-immunoreactive cells were detected, but few SPRR1/3-positive cells. Double staining demonstrated that cells containing SPRR1/3 almost always contained involucrin, but involucrin was present in many cells that did not contain SPRR. Light and electron microscopic immunohistochemistry of a stratified culture demonstrated that lower cells (close to the basal layer) were occasionally involucrin-positive, but lacked SPRR1/3, whereas more superficial cells contained both involucrin and SPRR. We conclude that involucrin and SPRR are sequentially induced in this order during keratinocyte differentiation.