Separation of Epidermal Layers of the Newborn Rat

Walker, Glenn K.; Sachs, L; Sibrack, Laurence A.; Ball, Richard D.; Bernstein, Isadore A.

doi:10.1111/1523-1747.ep12491663

Cited by 13 publications

(5 citation statements)

References 13 publications

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“…A single sample of cell envelopes was isolated from a pool of epidermal sheets obtained by this means. Samples of epidermis from skin obtained post mortem were prepared free of dermis by treatment with 0.24 M-NH4Cl at pH 9.5 as described [16]. Cell envelopes were isolated from epidermis, callus and psoriatic scales by gentle overnight stirring at room temperature in 0.025 M-Tris/HCl buffer, pH 9, contaihing 8 M-urea, 2 mM-2-mercaptoethanol and 0.2 % SDS, followed by centrifugation for 20 min at 750 g. The insoluble crude envelopes were freed of remaining soluble impurities by washing three times for 2 h with the same buffer mixture at room temperature and three times for 5 min with this mixture at 100 'C.…”

Section: Methodsmentioning

confidence: 99%

N 1 N 8-bis(γ-glutamyl)spermidine cross-linking in epidermal-cell envelopes. Comparison of cross-link levels in normal and psoriatic cell envelopes

Martinet

Beninati

Nigra

et al. 1990

Biochemical Journal

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N1N8-Bis(gamma-glutamyl)spermidine was found in exhaustive proteolytic digests of isolated cell envelopes from human epidermis at levels comparable with those of epsilon-(gamma-glutamyl)lysine. Significantly higher than normal amounts of these compounds, particularly the bis(gamma-glutamyl)polyamine, were observed in envelopes from afflicted areas (scales) of psoriatic patients. These findings support the notions that N1N8-bis(gamma-glutamyl)spermidine, like epsilon-(gamma-glutamyl)lysine, functions in cell envelopes as an enzyme-generated protein cross-link and stabilizing force and that individuals with the chronic, recurrent skin disease, psoriasis, exhibit in involved epidermis abnormal cell-envelope-protein cross-linking.

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Section: Methodsmentioning

confidence: 99%

N 1 N 8-bis(γ-glutamyl)spermidine cross-linking in epidermal-cell envelopes. Comparison of cross-link levels in normal and psoriatic cell envelopes

Martinet

Beninati

Nigra

et al. 1990

Biochemical Journal

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“…Postnatal back skins were collected from suckling mice at 2∫2, 6∫2, 18∫2, and 72∫2 h after birth. The epidermis was obtained by the alkaline ammonium chloride method of Walker et al (13) under cooling with ice, washed with ice cold PBS, and stored at ª76aeC until use.…”

Section: Animals and Tissue Samplesmentioning

confidence: 99%

Dynamic aspects of protein deimination in developing mouse epidermis

Akiyama

Senshu

1999

Experimental Dermatology

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The cornified layer of mammalian epidermis contains deiminated keratins and filaggrin whose arginine residues are partly converted to citrulline residues by peptidylarginine deiminase (EC 3.5.3.15). We have attempted to study dynamic aspects of protein deimination using late embryonic to early postnatal mouse skin. The epidermis was separated from the dermis by brief immersion of skin into a weakly alkaline ammonium chloride solution. The total homogenate of the epidermis was subjected to western blotting analyses for quantitative densitometry of major keratins, deiminated proteins and immunoreactive filaggrin. We found marked increases in both deiminated keratins and deiminated filaggrin from the 18th day of gestation to 2 h after birth followed by rapid decreases to minimum levels at 6 h and subsequent gradual increases surpassing the earlier levels by 72 h after birth. Such variations were associated with consistent changes of the intensity of deiminated proteins stained immunocytochemically. These results suggest that the protein deimination might play a role in dealing with the drastic environmental change after birth. Furthermore, we found compartmentalization of both total and deiminated filaggrins into soluble and particulate fractions. The soluble compartment contained relatively more deiminated filaggrin than the particulate fraction.

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“…may be correct. As Walkcr et al indicated in their recent paper ( 17), the incubation of rat skin in a butl er solution containing 0.5 '1-trypsin caused dissolution of the usual type of keratohyalin granule. lf this is correct and DH D are resistant to trypsin digestion.…”

Section: Discussionmentioning

confidence: 88%

The fluroescent profiles of keratohyalin granules of newborn rat epidermis with a new fluorescent thiol reagent (DACM)

Tezuka¹,

Hirai²,

Ogawa³

1978

Acta Derm Venereol

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A highly specific fluorescent thiol reagent, N-(7-Dimethylamino-4-methyl-coumarinyl)maleimide (DACM) has been used to examine the newborn rat skin in order to elucidate the cystine-rich keratohyalin granules or dense homogeneous deposits. After reduction of the disulphide bond with 2-mercaptoethanol, the periphery of the stratum corneum cells showed a very distinct fluorescence and the cytoplasm of the stratum corneum, granulosum and spinulosum showed moderate fluorescence. No fluorescence was observed, however, over the keratohyalin granules themselves. Intense, spotty fluorescence was observed from the remnants of nuclei and at the periphery of large keratohyalin granules of the stratum granulosum cells, which were assumed to be dense homogeneous deposts according to their distribution pattern. As the intense fluorescence elicited with DACM determines the abundance of cystine content in the material, cystine was found to be very scarce in keratohyalin granules, but abundant in dense homogeneous deposits and at the periphery of the stratum corneum cells.

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Separation of Epidermal Layers of the Newborn Rat

Cited by 13 publications

References 13 publications

N 1 N 8-bis(γ-glutamyl)spermidine cross-linking in epidermal-cell envelopes. Comparison of cross-link levels in normal and psoriatic cell envelopes

N 1 N 8-bis(γ-glutamyl)spermidine cross-linking in epidermal-cell envelopes. Comparison of cross-link levels in normal and psoriatic cell envelopes

Dynamic aspects of protein deimination in developing mouse epidermis

The fluroescent profiles of keratohyalin granules of newborn rat epidermis with a new fluorescent thiol reagent (DACM)

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