Presence in human epidermal cells of a soluble protein precursor of the cross-linked envelope: Activation of the cross-linking by calcium ions

Rice, Robert H.; Green, Howard

doi:10.1016/0092-8674(79)90123-5

Cited by 760 publications

(446 citation statements)

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“…As shown in Figure 4 (c and d), involucrin expression in the organotypic cultures was epibasal (red), as described for stratified cultures or hyperproliferative human skin. 12 EGFP expression (green) was observed in all layers of cultures seeded with LTR-EGFP-transduced keratinocytes indicating pan-epithelial expression of EGFP (Figure 4a and c). However, INV/LTR-directed EGFP expression co-localized with endogenous involucrin expression to suprabasal layers of the cultures (Figure 4b and d).…”

Section: Stratum-specific Expression Of Egfp In Epidermal Raft Culturesmentioning

confidence: 94%

“…The INV/LTR-GFP plasmid DNA was used as a control to demonstrate simultaneous amplification of both 5' LTR and 3' hybrid LTR when both are present (lane 8). The sequence of amplified fragments was confirmed by restriction analysis using XbaI (lanes [11][12][13].…”

Section: Construction Of Retroviral Vectors Targeted To the Upper Strmentioning

confidence: 99%

“…19 In cultures of human keratinocytes, involucrin is expressed by all cells that have left the basal layer. 12 To determine if transgene expression is confined to the upper strata of epidermis, organotypic cultures were constructed with keratinocytes transduced with retroviral vectors in which EGFP expression was directed by either the LTR or the INV/LTR hybrid promoter. Mature differentiated cultures were fixed and cryosectioned and expression of EGFP was examined by fluorescent microscopy.…”

Section: Stratum-specific Expression Of Egfp In Epidermal Raft Culturesmentioning

confidence: 99%

“…Involucrin is one of the precursor proteins used in assembling the keratinocyte-cornified envelope, and is specifically expressed in suprabasal layers of stratifying squamous epithelia. 12 A 2500-base fragment of hINV gene upstream regulatory region is sufficient to drive tissue-and stratum-specific expression in transgenic mice. 13,14 Deletion mapping studies have identified two regulatory regions that contribute to differentiation-induced activity of this promoter; a distal element (Ϫ2473/Ϫ2088) containing AP-1 and Sp1 binding sites, and a proximal region (Ϫ185/Ϫ1) with a critical AP-1 binding site overlapping with a consensus binding site for ets transcription factor.…”

Section: Introductionmentioning

confidence: 99%

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Durable and stratum-specific gene expression in epidermis

Ghazizadeh

Doumeng²,

Taichman³

2002

Gene Ther

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Section: Stratum-specific Expression Of Egfp In Epidermal Raft Culturesmentioning

confidence: 94%

Section: Construction Of Retroviral Vectors Targeted To the Upper Strmentioning

confidence: 99%

Section: Stratum-specific Expression Of Egfp In Epidermal Raft Culturesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Durable and stratum-specific gene expression in epidermis

Ghazizadeh

Doumeng²,

Taichman³

2002

Gene Ther

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“…In granular keratinocytes, proteins associated with the later stages of differentiation are expressed. These late differentiated markers include: profilaggrin (4), the precursor of the keratin cross-linking protein, filaggrin, transglutaminase I (5), and cornified envelope precursors such as involucrin (6) and loricrin (7). In corneocytes, loricrin and other structure proteins are extensively cross-linked by transglutaminase I to form the cornified envelope.…”

mentioning

confidence: 99%

The role of the calcium-sensing receptor in epidermal differentiation

et al. 2004

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Calcium regulates the proliferation and differentiation of keratinocytes both in vivo and in vitro. Elevated extracellular Ca 2+ concentration ([Ca 2+ ] o ) raises the intracellular free calcium ([Ca 2+ ] i ) and activates differentiation-related genes. Cells lacking the calcium sensing receptor (CaR) fail to respond to [Ca 2+ ] o and to differentiate, indicating a role for CaR in keratinocyte differentiation. These concepts derived from in vitro experiments have been tested and confirmed in two mouse models. 4 CALCIUM IS A KEY REGULATOR FOR EPIDERMAL DIFFERENTIATIONThe mammalian epidermis is a highly specialized, highly organized, stratified squamous epithelium consisting of basal, spinous, granular and cornified cell layers. Each layer is defined by distinguishing morphological and biochemical features and state of differentiation of the keratinocytes (1). While cell proliferation occurs only in basal layer, post-mitotic keratinocytes undergo progressive differentiation as they travel across the epidermis towards the stratum corneum. Keratinocytes in each epidermal layer express distinct biochemical markers. Keratin 5 (K5) and 14 (K14) are predominantly expressed in basal keratinocytes (2). In spinous cells, synthesis of two early differentiation markers, keratins K1 and K10, is initiated (2, 3). In granular keratinocytes, proteins associated with the later stages of differentiation are expressed. These late differentiated markers include: profilaggrin (4), the precursor of the keratin cross-linking protein, filaggrin, transglutaminase I (5), and cornified envelope precursors such as involucrin (6) and loricrin (7). In corneocytes, loricrin and other structure proteins are extensively cross-linked by transglutaminase I to form the cornified envelope. Calcium-induced Keratinocyte differentiationPrimary keratinocytes provide an excellent in vitro model for studying epidermal differentiation (8 level in the outer epidermis, independent of barrier perturbation, directly regulate lamellar body secretion and the expression of differentiation-specific markers in vivo (27,28). unlike the full-length CaR ( Fig. 2A) which displays distinct plasma membrane localization, the distribution of spliced variant (Fig. 2B) is strictly confined in the cytoplasm with a perinuclear pattern. Failure to generate a protein capable of being transferred to the cell surface due to a change in glycosylation is a likely basis for the failure of the CaR spliced variant to mediate cellular response to [Ca] o . Calcium signaling in the keratinocytesThe full-length CaR and its spliced variant are expressed differentially during keratinocyte differentiation. In Western analyses of keratinocyte membrane proteins (Fig. 3A) the endogenous CaR was detected by an antibody (ADDR) that reacts with both forms of CaR as three major bands of 120-kDa, 160-kDa and 185-kDa and a minor band of 130-kDa (which was best seen in longer exposed film). Whereas the 120-kDa band corresponds to the nonglycosylated full-length CaR, the 160-kDa band co...

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