DOCSLIB.ORG

Expression of Trichohyalin in Dermatological Disorders: a Comparative Study with Involucrin and Filaggrin by Immunohistochemical Staining

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Expression of Trichohyalin in Dermatological Disorders: a Comparative Study with Involucrin and Filaggrin by Immunohistochemical Staining Acta Derm Venereol (Stockh) 1999; 79: 122^126 Expression of Trichohyalin in Dermatological Disorders: a Comparative Study with Involucrin and Filaggrin by Immunohistochemical Staining SEUNG-CHUL LEE1,2, JEE-BUM LEE1, JAE-JEONG SEO1 and YOUNG PIO KIM1 1Department of Dermatology, Chonnam University Medical School and 2Research Institute of Medical Sciences, Chonnam University, Kwangju, South Korea To investigate the function of trichohyalin during terminal In the present study, an attempt was made to delineate the di¡erentiation of the skin, immunohistochemical studies were role of trichohyalin as a component of the CE during terminal performed on trichohyalin and its related proteins, ¢laggrin di¡erentiation by screening its expression in various skin disor- and involucrin, the components of the corni¢ed cell envelope. ders. We also evaluated the expression of trichohyalin in com- In skin disorders unrelated to tumours, weak trichohyalin parison with that of ¢laggrin and involucrin to clarify the expression was found in a few granular cells or in the horny functional relationship among them as precursor proteins of layer of psoriasis, ichthyosis, keratosis pilaris, porokeratosis, the CE. chronic dermatitis and callus. Similar trichohyalin expression was found in epidermal tumours, such as seborrheic keratosis, actinic keratosis, Bowen's disease and well-di¡erentiated squamous cell carcinoma. In follicular tumours, trichohyalin MATERIALS AND METHODS expression was positive in trichoepithelioma, keratotic basal Materials cell epithelioma, proliferating trichilemmal tumour, trichi- Formalin-¢xed, para¤n-embedded samples were selected after evalua- lemmoma, pilomatricoma and keratoacanthoma. From com- tion of their histopathological features with haematoxylin and eosin parative studies with ¢laggrin and involucrin, trichohyalin staining. In the present study, 49 samples of skin disorders unrelated expression was co-localized with them in molluscum contagio- to tumours, 36 samples of epidermal tumours and 25 samples of sum, keratoacanthoma and pilomatricoma. From this study, follicular tumours were examined. trichohyalin is revealed to have close functional relationship with other markers of terminal di¡erentiation as a precursor Antibodies of the corni¢ed cell envelope of the skin. Key words: terminal A polyclonal anti-trichohyalin antibody was elicited from rabbits with di¡erentiation; corni¢ed cell envelope; precursor. recombinant human trichohyalin of 1,250 ^ 1,849 residues (domain 8) (Accepted September 7, 1998.) located in the carboxy-terminus (16). Antibodies to involucrin and Acta Derm Venereol (Stockh) 1999; 79: 122^126. ¢laggrin were purchased from Biomedical Technologies Inc. (MA, USA). The polyclonal anti-involucrin antibody was raised from rabbits Seung-Chul Lee, Department of Dermatology, Chonnam with puri¢ed cultured human epidermal cells (17). The monoclonal University Medical School, 8 Hak-dong, Tong-gu, Kwangju anti-¢laggrin antibody was elicited from mice with partially puri¢ed 501 ^ 190, South Korea. ¢laggrin of the newborn human epidermis (18). Immunohistochemical staining of trichohyalin, involucrin and During terminal di¡erentiation of the skin, a characteristic ¢laggrin structure called the corni¢ed cell envelope (CE) is formed in Immunohistochemical procedures were carried out according to the the inner surface of the cell periphery of the granular layer. It protocol described in the LSAB kit (DAKO, CA, USA) with minor is a rigid, complex structure composed of many precursor pro- modi¢cations. Samples 5 mm thick were depara¤nized, rehydrated teins of transglutaminase (TGase) which catalyses e-(c-gluta- and pre-treated with 3% hydrogen peroxide for 10 min to block endo- myl)lysine cross-linking (1 ^ 7). Trichohyalin is one of genous peroxidase activity, followed by incubation in the blocking substrate proteins of TGase, and it can bind with keratin ¢la- reagent for 10 min. Then the sections were incubated with the primary ments as intermediate ¢lament associated protein (IFAP) (8, antibodies under the following conditions: anti-trichohyalin at 1 : 50 9). Trichohyalin is found mainly in the inner root sheath dilution for 1 h; anti-involucrin at 1 : 10 dilution for 30 min; and anti- (IRS) and medulla of the hair follicle where it gives a structural ¢laggrin at 1 : 150 dilution for 30 min at room temperature. For tricho- rigidity, but its possible function as a precursor of the CE has hyalin staining, sectioned samples were pre-treated in the autoclave at been proposed previously (8). In a recent report (10), tricho- 121³C, 15 lbs/inch2 for 5 min before reaction with the primary anti- hyalin is found as a cross-linked component of mouse foresto- body to retrieve antigenicity. Then the sections were incubated with a mach CE, and it is thought to modulate the biomechanical biotin-labelled anti-immunoglobulin antiserum for 15 min, followed by peroxidase-conjugated streptoavidin for 15 min. As the chromogen, properties of the CE. In the skin, however, trichohyalin is £eet- 3-amino-9-ethylcarbazole, was applied, red-coloured precipitates ingly expressed in the neonatal foreskin, and it is not detected appeared at the antigen site. During immunohistochemical staining, in the adult epidermis (11, 12). A few scattered observations each sample was washed with phosphate-bu¡ered saline (pH 7.4). Sec- reported that it is aberrantly expressed only in a number of der- tions were counterstained with Meyer's haematoxylin and mounted matoses, such as psoriasis, molluscum contagiosum, actinic with Universal Mount (Research Genetics, AL, USA). As a negative keratosis, trichofolliculoma, epidermolytic hyperkeratosis control, sections were incubated with non-immunized serum instead and pilomatricoma (13 ^ 15). of the primary antibodies. Acta Derm Venereol (Stockh) 79 # 1999 Scandinavian University Press. ISSN 0001-5555 Trichohyalin expression in dermatological disorders 123 Fig. 1. Localization of trichohyalin in (a) ichthyosis vulgaris (6160), (b) Bowen's disease (6200), (c) trichoepithelioma (6200) and (d) keratotic basal cell epithelioma (6200). Arrows and arrowheads indicate positive expression of trichohyalin. RESULTS keratosis and SCC, several dyskeratotic cells were also positive in trichohyalin expression. No immunoreactivity was found in Trichohyalin expression in skin disorders unrelated to malignant tumours of non-keratotic basal cell carcinoma tumours (BCC). In depara¤nized autoclaved sections, trichohyalin expression was not detected in the interfollicular epidermis of normal adult skin, but it was strongly expressed in the IRS and medulla Trichohyalin expression in follicular tumours of hair follicles (data not shown). Such immunoreactivity was Immunoreactivity of trichohyalin was found in trichoepithe- not detected in the sections without pre-treatment by autoclav- lioma (Fig. 1c), keratotic BCC (Fig. 1d), proliferating tri- ing. Among samples, weak immunoreactivity of trichohyalin chilemmal tumour, trichilemmoma, pilomatricoma and was localized to a few granular cells or to the horny layer of keratoacanthoma. In trichoepithelioma and keratotic BCC, ichthyosis (Fig. 1a), psoriasis, keratosis pilaris, porokeratosis, immunoreactivity was found around horn cysts, but there was chronic dermatitis and callus. Strong immunreactivity of tri- contrast between them; major immunoreactivity was localized chohyalin was found from the horny to spinous layers of mol- around cysts showing a palisading pattern in trichoepithe- luscum contagiosum (Fig. 2b). lioma, while positive immunoreactivity was localized to cystic contents in keratotic BCC. In pilomatricoma, immunoreactiv- ity of trichohyalin was localized mainly to the junction between Trichohyalin expression in epidermal tumours basophilic and shadow cells (Fig. 2h). The expression pattern in several epidermal tumours, such as seborrheic keratosis, actinic keratosis, Bowen's disease Sequential expression of trichohyalin, ¢laggrin and involucrin (Fig. 1b) and squamous cell carcinoma (SCC), was similar to in molluscum contagiosum, keratoacanthoma and that of skin disorders unrelated to tumours, as described pilomatricoma before. In particular, all samples of seborrheic keratosis showed positive expression of trichohyalin in the granular In a comparative study of trichohyalin with ¢laggrin and and horny layers, and in the lining cells of horn cysts. In actinic involucrin, we found a sequential order among them in 3 Acta Derm Venereol (Stockh) 79 124 S.-C. Lee et al. Fig. 2. Co-localization of (b, e, h) trichohyalin with (c, f, i) ¢laggrin and (a, d, g) involucrin in molluscum contagiosum (a ^ c, 6100), keratoa- canthoma (d ^ f, 6132) and pilomatricoma (g ^ i, 680). dermatoses. In molluscum contagiosum, major immunoreac- and follicular origins. Such morphological evidence strongly tivity of involucrin was localized to the upper or mid-spinous suggests a functional role for trichohyalin as a precursor of layer, while that of ¢laggrin was con¢ned to the horny layer of the CE in the keratinizing process of the skin. Notably, tricho- molluscum body (Fig. 2a, c). Immunoreactivity of trichohyalin hyalin was co-localized with the other precursors of the CE, was scattered to whole layers, from the horny to upper spinous involucrin and ¢laggrin, in molluscum contagiosum and in fol- layers, overlapping with the other precursors (Fig. 2b). A simi- licular-originating tumours, such as keratoacanthoma and lar pattern of immunoreactivity was found in the follicular- pilomatricoma. The overlapped
Load more

Recommended publications
  • Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino-Like Mouse Models: Tracking the Role of the Hairless Gene
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 5-2006 Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino-like Mouse Models: Tracking the Role of the Hairless Gene Yutao Liu University of Tennessee - Knoxville Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Part of the Life Sciences Commons Recommended Citation Liu, Yutao, "Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino- like Mouse Models: Tracking the Role of the Hairless Gene. " PhD diss., University of Tennessee, 2006. https://trace.tennessee.edu/utk_graddiss/1824 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Yutao Liu entitled "Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino-like Mouse Models: Tracking the Role of the Hairless Gene." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Doctor of Philosophy, with a major in Life Sciences. Brynn H. Voy, Major Professor We have read this dissertation and recommend its acceptance: Naima Moustaid-Moussa, Yisong Wang, Rogert Hettich Accepted for the Council: Carolyn R.
    [Show full text]
  • Differential Roles of P63 Isoforms in Epidermal Development: Selective Genetic Complementation in P63 Null Mice
    Cell Death and Differentiation (2006) 13, 1037–1047 & 2006 Nature Publishing Group All rights reserved 1350-9047/06 $30.00 www.nature.com/cdd Differential roles of p63 isoforms in epidermal development: selective genetic complementation in p63 null mice E Candi1, A Rufini1, A Terrinoni1, D Dinsdale2, M Ranalli1, The skin consists of two compartments, the dermis and the A Paradisi1, V De Laurenzi2, LG Spagnoli1, MV Catani1, epidermis. The latter is a multilayered, stratified epithelium S Ramadan1, RA Knight2 and G Melino*,1,2 continuously regenerated by terminally differentiating keratino- cytes, a process called cornification1–3 (Figure 1a and b). 4 1 Biochemistry Laboratory, IDI-IRCCS, c/o University of Rome ‘Tor Vergata’, Recent evidence demonstrates a major role for p63, a 00133 Rome, Italy member of the p53 family,5–8 in this process as mutations in 2 Medical Research Council, Toxicology Unit, Leicester University, Leicester the TP63 gene cause limb and skin defects in humans,9 and LE1 9HN, UK p63À/À mice have no epidermis, no limbs and die at birth * Corresponding author: G Melino, Medical Research Council, Toxicology Unit, owing to dehydration.10,11 Hodgkin Building, Leicester University, Lancaster Road, PO Box 138, Leicester LE1 9HN, UK. Tel: þ 44 116 252 5616; Fax: þ 44 116 252 5551; The expression of p63 proteins originates from two E-mail: [email protected] promoters, giving rise to TAp63 and DNp63 isoforms. In addition, both isoforms undergo alternative splicing at the Received 03.3.06; revised 08.3.06; accepted 08.3.06; published online 07.4.06 C-terminus producing three different TAp63 and DNp63 Edited by P Vandenabeele isoforms (a, b and g).
    [Show full text]
  • Loss-Of-Function Mutations in the Gene Encoding Filaggrin Cause Ichthyosis
    LETTERS Loss-of-function mutations in the gene encoding filaggrin cause ichthyosis vulgaris Frances J D Smith1, Alan D Irvine2, Ana Terron-Kwiatkowski1, Aileen Sandilands1, Linda E Campbell1, Yiwei Zhao1, Haihui Liao1, Alan T Evans3, David R Goudie4, Sue Lewis-Jones5, Gehan Arseculeratne5, Colin S Munro6, Ann Sergeant6, Gra´inne O’Regan2, Sherri J Bale7, John G Compton7, John J DiGiovanna8,9, Richard B Presland10,11, Philip Fleckman11 & W H Irwin McLean1 Ichthyosis vulgaris (OMIM 146700) is the most common composed of the 400-kDa protein profilaggrin. Following a short, inherited disorder of keratinization and one of the most unique N-terminal domain, most of the profilaggrin molecule consists frequent single-gene disorders in humans. The most widely of 10–12 repeats of the 324-residue filaggrin sequence6. Upon terminal cited incidence figure is 1 in 250 based on a survey of differentiation of granular cells, profilaggrin is proteolytically cleaved 1 B http://www.nature.com/naturegenetics 6,051 healthy English schoolchildren . We have identified into 37-kDa filaggrin peptides and the N-terminal domain contain- homozygous or compound heterozygous mutations R501X and ing an S100-like calcium binding domain. Filaggrin rapidly aggregates 2282del4 in the gene encoding filaggrin (FLG) as the cause of the keratin cytoskeleton, causing collapse of the granular cells into moderate or severe ichthyosis vulgaris in 15 kindreds. In flattened anuclear squames. This condensed cytoskeleton is cross- addition, these mutations are semidominant; heterozygotes linked by transglutaminases during formation of the cornified cell show a very mild phenotype with incomplete penetrance. envelope (CCE). The CCE is the outermost barrier layer of the skin The mutations show a combined allele frequency of B4% which not only prevents water loss but also impedes the entry of in populations of European ancestry, explaining the high allergens and infectious agents7.
    [Show full text]
  • An Abridged Compendium of Words. a Discussion of Them and Opinions About Them
    DERMATOLOGY PRACTICAL & CONCEPTUAL www.derm101.com Dermatopathology: An abridged compendium of words. A discussion of them and opinions about them. Part 6 (I-L) Bruce J. Hookerman1 1 Dermatology Specialists, Bridgeton, Missouri, USA Citation: Hookerman BJ. Dermatopathology: An abridged compendium of words. A discussion of them and opinions about them. Part 6 (I-L). Dermatol Pract Concept. 2014;4(4):1. http://dx.doi.org/10.5826/dpc.0404a01 Copyright: ©2014 Hookerman. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Corresponding author: Bruce J. Hookerman, M.D., 12105 Bridgeton Square Drive, St. Louis, MO 63044, USA. Email: [email protected] – I – term “id reaction” only for a spongiotic dermatitis manifested by tiny vesicles on the hands of patients with florid dermato- ICHTHYOSIS: a generic term for skin conditions character- phytosis at another site, usually the feet, or for an analogue ized by what are said to be fishlike scales, i.e., scales that are of that phenomenon such as widespread vesicles that appear broad and polygonal with free edges, as are seen in ichthyosis subsequent to injudicious treatment, i.e., with Gentian violet vulgaris (and its look-alike, acquired ichthyosis), X-linked (known sardonically in times past as “Gentian violent”) of ichthyosis, and lamellar ichthyosis. Conditions reputed to be an exuberant spongiotic dermatitis, usually on the feet, such ichthyosis, such as ichthyosis hystrix and ichthyosis linearis as an allergic contact dermatitis. A time-honored explana- circumflexa, do not qualify because they are not associated tion for an “id” reaction is hematogenous dissemination of with broad polygonal scales.
    [Show full text]
  • Keratinization of the Oral Epithelium
    Annals of the Royal College of Surgeons of England (I976) vol 58 Keratinization of the oral epithelium David Adams BSC MDS PhD Department of Oral Biology, Welsh National School of Medicine Dental School, Cardiff Summary compare it with the non-keratinizing mucosa The morphology of the keratinizing epi- and the epidermis. I also wish to examine thelia in the mouth is reviewed in the light of some of the mechanisms which control and recent knowledge. There appears to be a spec- regulate keratinization and discuss briefly the trum of degrees of keratinization rather than clinical implications of these. distinct types, and the degree of keratinization Orthokeratinization, in which the surface is reflected in the degree of packing and orien- undergoes cornification as cells lose their stain- tation of tonofilaments. The role of keratohya- ing characteristics and their nuclei, is found on line and other granules in the process is dis- the hard palate and on gingiva, especially cussed and it is suggested that modification where this is firmly bound down to of the cell membrane is an important part of underlyinlg bone (Fig. i). The epithelium keratinization. Although the potential of the lies on a basement membrane which separates various areas in the mucosa is genetically de- it from the connective tissue. Above this there termined and appears early in fetal life, the is a series of more or less well-defined layers. connective tissue exerts an influence on the First comes the germinal layer, one or two extent of keratinization of the surface in a cells thick, then the stratum spinosum, with manner which is not understood.
    [Show full text]
  • Perspectives on Morphologic Approaches to the Study of the Granular Layer Keratinocyte Karen A
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biologic Structure and Function: Perspectives on Morphologic Approaches to the Study of the Granular Layer Keratinocyte Karen A. Holbrook, Ph.D. Stephen Rothman [1] wrote: ‘‘Modern dermatology has been built on the of morphologic approaches to understand skin biology, selecting as solid pillars of precise macro- and microscopic observation, accurate an illustration the progress that has been made in understanding recording and meaningful interpretation of the findings. No natural the structure, composition and function of one cell, the granular layer science can exist without direct observation of natural phenomena, and keratinocyte. This approach avoids describing the advances in metho- dermatology has become a great discipline because we have had so dology, and, instead, demonstrates how morphology has promoted the many good observers.’’ He went on to state further that ‘‘y whereas the conceptual development of the topic (Tables I–III). immense knowledge acquired by the classical descriptive methods is still rapidly increasing, the application of experimental methods to derma- WHAT DO MORPHOLOGIC METHODS OFFER? tologic problems is a relatively young and undeveloped approach.’’ AN OVERVIEW These statements acknowledge the importance of observation and The development of morphologic approaches for skin research has description (usually thought of as features of morphology) in clinical involved the development of instrumentation (to a large extent, dermatology and in descriptive dermatologic research, but were made microscopy) methods to prepare tissue for morphologic examination before he could recognize the value of morphology to experimental (e.g., biopsy, separated epidermis and dermis, isolated cells, cell studies as well.
    [Show full text]
  • Deimination, Intermediate Filaments and Associated Proteins
    International Journal of Molecular Sciences Review Deimination, Intermediate Filaments and Associated Proteins Julie Briot, Michel Simon and Marie-Claire Méchin * UDEAR, Institut National de la Santé Et de la Recherche Médicale, Université Toulouse III Paul Sabatier, Université Fédérale de Toulouse Midi-Pyrénées, U1056, 31059 Toulouse, France; [email protected] (J.B.); [email protected] (M.S.) * Correspondence: [email protected]; Tel.: +33-5-6115-8425 Received: 27 October 2020; Accepted: 16 November 2020; Published: 19 November 2020 Abstract: Deimination (or citrullination) is a post-translational modification catalyzed by a calcium-dependent enzyme family of five peptidylarginine deiminases (PADs). Deimination is involved in physiological processes (cell differentiation, embryogenesis, innate and adaptive immunity, etc.) and in autoimmune diseases (rheumatoid arthritis, multiple sclerosis and lupus), cancers and neurodegenerative diseases. Intermediate filaments (IF) and associated proteins (IFAP) are major substrates of PADs. Here, we focus on the effects of deimination on the polymerization and solubility properties of IF proteins and on the proteolysis and cross-linking of IFAP, to finally expose some features of interest and some limitations of citrullinomes. Keywords: citrullination; post-translational modification; cytoskeleton; keratin; filaggrin; peptidylarginine deiminase 1. Introduction Intermediate filaments (IF) constitute a unique macromolecular structure with a diameter (10 nm) intermediate between those of actin microfilaments (6 nm) and microtubules (25 nm). In humans, IF are found in all cell types and organize themselves into a complex network. They play an important role in the morphology of a cell (including the nucleus), are essential to its plasticity, its mobility, its adhesion and thus to its function.
    [Show full text]
  • Cell Shape Controls Terminal Differentiation of Human Epidermal Keratinocytes (Cell Adhesion/Proliferation/Involucrin Expression) FIONA M
    Proc. Natl. Acad. Sci. USA Vol. 85, pp. 5576-5580, August 1988 Cell Biology Cell shape controls terminal differentiation of human epidermal keratinocytes (cell adhesion/proliferation/involucrin expression) FIONA M. WATT*, PETER W. JORDANt, AND CHARLES H. O'NEILLt *Keratinocyte Laboratory, and tAnchorage Laboratory, Imperial Cancer Research Fund, P.O. Box 123, Lincoln's Inn Fields, London WC2A 3PX, United Kingdom Communicated by Howard Green, May 2, 1988 ABSTRACT Cultures of human epidermal keratinocytes To investigate the role of cell-substratum contact, and provide a useful experimental model with which to study the hence cell shape, in regulating terminal differentiation, we factors that regulate cell proliferation and terminal differen- have made use of a recently described technique that allows tiation. One situation that is known to trigger premature precise control of cell-substratum contact in the absence of terminal differentiation is suspension culture, when keratin- cell-cell contact (14). We have looked at the effect ofchanges ocytes are deprived of substratum and intercellular contact. in cell shape on DNA synthesis and involucrin expression We have now investigated whether area ofsubstratum contact, and conclude that a rounded morphology acts as a signal to and hence cell shape, can regulate terminal differentiation. keratinocytes to stop dividing and to undergo terminal Keratinocytes were grown on circular adhesive islands that differentiation; inhibition ofproliferation in spread cells is not prevented cell-cell contact. By varying island area we could a sufficient signal for terminal differentiation. vary cell shape from fully spread to almost spherical. We found that when substratum contact was restricted, DNA synthesis was inhibited and expression of involucrin, a marker of MATERIALS AND METHODS terminal differentiation, was stimulated.
    [Show full text]
  • Identification of Trichoplein, a Novel Keratin Filament- Binding Protein
    Research Article 1081 Identification of trichoplein, a novel keratin filament- binding protein Miwako Nishizawa1,*, Ichiro Izawa1,*, Akihito Inoko1,*, Yuko Hayashi1, Koh-ichi Nagata1, Tomoya Yokoyama1,2, Jiro Usukura3 and Masaki Inagaki1,‡ 1Division of Biochemistry, Aichi Cancer Center Research Institute, 1-1 Kanokoden, Chikusa-ku, Nagoya 464-8681, Japan 2Department of Dermatology, Mie University Faculty of Medicine, 2-174 Edobashi, Tsu 514-8507, Japan 3Department of Anatomy and Cell Biology, Nagoya University School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-8550, Japan *These authors contributed equally to this work ‡Author for correspondence (e-mail: [email protected]) Accepted 29 November 2004 Journal of Cell Science 118, 1081-1090 Published by The Company of Biologists 2005 doi:10.1242/jcs.01667 Summary Keratins 8 and 18 (K8/18) are major components of the antibody in a complex with K8/18 and immunostaining intermediate filaments (IFs) of simple epithelia. We report revealed that trichoplein colocalized with K8/18 filaments here the identification of a novel protein termed in HeLa cells. In polarized Caco-2 cells, trichoplein trichoplein. This protein shows a low degree of sequence colocalized not only with K8/18 filaments in the apical similarity to trichohyalin, plectin and myosin heavy chain, region but also with desmoplakin, a constituent of and is a K8/18-binding protein. Among interactions desmosomes. In the absorptive cells of the small intestine, between trichoplein and various IF proteins that we trichoplein colocalized with K8/18 filaments at the apical tested using two-hybrid methods, trichoplein interacted cortical region, and was also concentrated at desmosomes.
    [Show full text]
  • Keratinization and Its Disorders
    Oman Medical Journal (2012) Vol. 27, No. 5: 348-357 DOI 10. 5001/omj.2012.90 Review Article Keratinization and its Disorders Shibani Shetty, Gokul S. Received: 03 May 2012 / Accepted: 08 July 2012 © OMSB, 2012 Abstract Keratins are a diverse group of structural proteins that form the epithelium (buccal mucosa, labial mucosa) and specialized intermediate filament network responsible for maintaining the mucosa (dorsal surface of the tongue).2 An important aspect structural integrity of keratinocytes. In humans, there are around of stratified squamous epithelia is that the cells undergo a 30 keratin families divided into two groups, namely, acidic and terminal differentiation program that results in the formation basic keratins, which are arranged in pairs. They are expressed in of a mechanically resistant and toughened surface composed of a highly specific pattern related to the epithelial type and stage of cornified cells that are filled with keratin filaments and lack nuclei cellular differentiation. A total of 54 functional genes exist which and cytoplasmic organelles. In these squames, the cell membrane codes for these keratin families. The expression of specific keratin is replaced by a proteinaceous cornified envelope that is covalently genes is regulated by the differentiation of epithelial cells within cross linked to the keratin filaments, providing a highly insoluble the stratifying squamous epithelium. Mutations in most of these yet flexible structure that protects the underlying epithelial cells.1 genes are now associated with specific tissue fragility disorders Keratinization, also termed as cornification, is a process which may manifest both in skin and mucosa depending on the of cytodifferentiation which the keratinocytes undergo when expression pattern.
    [Show full text]
  • The Human Involucrin Gene Contains Spatially Distinct Regulatory Elements That Regulate Expression During Early Versus Late Epidermal Di€Erentiation
    Oncogene (2002) 21, 738 ± 747 ã 2002 Nature Publishing Group All rights reserved 0950 ± 9232/02 $25.00 www.nature.com/onc The human involucrin gene contains spatially distinct regulatory elements that regulate expression during early versus late epidermal dierentiation James F Crish1, Frederic Bone1, Eric B Banks1 and Richard L Eckert*,1,2,3,4,5 1Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, Ohio, OH 44106-4970, USA; 2Department of Dermatology, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, Ohio, OH 44106-4970, USA; 3Department of Reproductive Biology, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, Ohio, OH 44106-4970, USA; 4Department of Biochemistry, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, Ohio, OH 44106-4970, USA; 5Department of Oncology, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, Ohio, OH 44106-4970, USA Human involucrin (hINV) is a keratinocyte protein that Keywords: transgenic mice; activator protein-1; AP1; is expressed in the suprabasal compartment of the jun; fos; Sp1; gene regulation; epidermal keratinocyte; epidermis and other stratifying surface epithelia. In- keratinocyte dierentiation; tissue speci®c volucrin gene expression is initiated early in the dierentiation process and is maintained until terminal cell death. The distal regulatory region (DRR) is a Introduction segment of the hINV promoter (nucleotides 72473/ 71953) that accurately recapitulates the normal pattern The major cell type, and the cell type responsible for of suprabasal (spinous and granular layer) expression in the morphological characteristics of the epidermis, is transgenic mouse epithelia.
    [Show full text]
  • Characterization of the Human Involucrin Promoter Using a Transient -Galactosidase Assay
    Journal of Cell Science 103, 925-930 (1992) 925 Printed in Great Britain © The Company of Biologists Limited 1992 Characterization of the human involucrin promoter using a transient -galactosidase assay JOSEPH M. CARROLL1 and LORNE B. TAICHMAN2,* 1Graduate Program in Cellular and Developmental Biology, 2Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8702, USA *Author for correspondence Summary Involucrin, a component of the cornified cell envelope, nascent RNA and suggested that sequences within the is expressed specifically in differentiating keratinocytes intron have regulatory activity. These results suggest of stratified squamous epithelia. To explore the regula- that the involucrin intron operates in vivo to regulate tion of involucrin expression, 3.7 kb of upstream expression in the epidermis. sequences of the human involucrin gene was cloned into a plasmid containing a -galactosidase reporter gene and transfected into early passage keratinocytes and a Abbreviations used: ADH, alcohol dehydrogenase; b-gal, b- galactosidase; DME, Dulbecco’s modified Eagle’s medium; variety of human cell types. The full-length construct DDAB, dimethyldioctyldecylammonium bromide; FCS, fetal calf gave maximal and tissue-specific expression. Deletion serum; kb, kilobase; ONPG, O-nitrophenyl b-D- analysis showed that sequences between 900 and 2500 galactopyranoside; PBS, phosphate buffered saline without bp upstream of the transcriptional start site and the calcium/magnesium salts; PCR, polymerase chain reaction; intron located between the transcriptional and transla- PtdEtn, dioleoyl-L-a -phosphatidylethanolamine; RSV, Rous tional start sites were required for maximal expression. sarcoma virus; SV40, simian virus 40.
    [Show full text]