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Johanna M Brandner

Publications and source records attributed to Johanna M Brandner.

10 recordsLinked to original sources
PubMed2005

Differential induction of connexins 26 and 30 in skin tumors and their adjacent epidermis.

Gap junctions (GJs) have been shown to play a role in tumor progression including a variety of keratinocyte-derived and non-keratinocyte-derived skin tumors. Here we show that the synthesis of the GJ proteins connexin 26 and connexin 30 (Cx26 and Cx30) is induced in keratinocyte-derived epithelial skin tumors whereas there is either no change or a downregulation of Cx43. Cx26, Cx30, and Cx43 are absent in non-epithelial skin tumors. Further, Cx26 and Cx30 are induced in the epidermis adjacent to malignant melanoma but absent in the epidermis adjacent to benign non-epithelial skin lesions (melanocytic nevi and angioma). The keratinocyte-derived skin tumors are very heterogeneous regarding the Cx26/Cx30 pattern in the epidermis at the periphery of the tumors. We did not observe any difference in the localization of the very similar proteins Cx26 and Cx30 but a variation in intensity of immunoreactivity. As the staining patterns of Cx26 and Cx30 antibodies are not identical to those of CK6, a marker for hyperproliferation, and CK17, a marker for trauma, we discuss that the induction of these gap junctional proteins exceeds a reflection of reactive hyperproliferative or traumatized epidermis. We further discuss the putative roles of these gap junctional proteins in tumor progression.

Animals↗
PubMed2005

In vitro isolation and cell culture of vestibular inner ear melanocytes.

INTRODUCTION: Melanocytes of the membranous labyrinth of the inner ear have been described morphologically in various contexts. Nature and functions of these cells are as yet not completely clear, even though several hypotheses exist regarding the same. The limited knowledge is due in part to a lack of methods regarding in vitro cell culture. The aim of this study was to describe conditions for the successful cell culture of vestibular inner ear melanocytes (VIEM), to compare their growth properties with those of epidermal melanocytes, and to characterize them immunohistochemically. MATERIALS AND METHODS: Membranous labyrinth cells from freshly slaughtered sheep were isolated, and melanocytes and fibroblasts subsequently cultured. In addition, melanocytes from the skin of the same sheep were cultured. Antibodies specific to tyrosinase, tyrosinase-related protein 1 (TRP-1/Mel-5), and melanoma-specific antigen A (Melan A) were used to analyze the cultured cells. RESULTS: The proliferation of VIEM was retarded in comparison to epidermal melanocytes. After 14 days, VIEM began to proliferate for the first time, whereas epidermal melanocytes proliferated already after 7 days. In contrast to epidermal melanocytes, the culturing process of VIEM seemed to be dependent on the presence of fibroblasts, and VIEM often accumulated in the vicinity of fibroblasts forming three-dimensional clusters. Moreover, VIEM showed a higher ratio of highly pigmented cells with a round cell shape and small dendrites in comparison to epidermal melanocytes. Immunohistochemical techniques proved the VIEM to be positive for Melan A, TRP-1 and, in the majority of cases, also for tyrosinase. CONCLUSION: We successfully cultured melanocytes of the inner ear vestibular labyrinth for the first time and demonstrated melanocytic characteristics of these cells. This accomplishment will provide the opportunity to investigate VIEM in more detail in future experiments.

Animals↗
PubMed2005

Human Merkel cells--aspects of cell biology, distribution and functions.

Human Merkel cells were first described by Friedrich S. Merkel in 1875 and named "Tastzellen" (touch cells) assuming a sensory touch function within the skin. Only ultrastructural research revealed their characteristics such as dense-core granules, plasma membrane spines and dendrites as well as a loosely arranged cytoskeleton. Biochemical analysis identified the expression of very specific cytokeratins (most notably CK 20) allowing the immunohistochemical detection of Merkel cells. In humans, they occur within the basal epidermis, being concentrated in eccrine glandular ridges of glabrous skin and in Haarscheiben of hairy skin, within belt-like clusters of hair follicles, and in certain mucosal tissues. Within the human skin, the dense-core granules contain heterogeneously distributed neuropeptides, some of which might work as neurotransmitters through which Merkel cells and their associated nerves exert their classical function as slowly adapting mechanoreceptors type I. This is the case in the Haarscheiben, small sensory organs containing keratinocytes with a special program of differentiation that includes the expression of CK 17 and Ber-EP4. Other peptides may act as growth factors and thus might participate in growth, differentiation and homeostasis of cutaneous structures. It is not yet clear whether the Merkel cell carcinomas, aggressive skin carcinomas, indeed arise from Merkel cells. We summarize and discuss data on the distribution, function and heterogeneity of human Merkel cells in normal and diseased skin.

Adult↗
PubMed2005

Up-regulated expression of zonula occludens protein-1 in human melanoma associates with N-cadherin and contributes to invasion and adhesion.

During the process of malignant transformation, nascent melanoma cells escape keratinocyte control through down-regulation of E-cadherin and instead communicate among themselves and with fibroblasts via N-cadherin-based cell-cell contacts. The zonula occludens (ZO) protein-1 is a membrane-associated component of both the tight and adherens junctions found at sites of cell-cell contact. In most cancers, levels of ZO-1 are typically down-regulated, leading to increased motility. Here we report the novel observation that ZO-1 expression is up-regulated in melanoma cells and is located at adherens junctions between melanoma cells and fibroblasts. Immunofluorescence and co-immunoprecipitation studies showed co-localization of ZO-1 with N-cadherin. Down-regulation of ZO-1 in melanoma cells through RNA interference produced marked changes in cell morphology--leading to a less-dendritic, more rounded phenotype. Consistent with a role in N-cadherin-based adhesion, RNAi-treated melanoma cells were less adherent and invasive when grown in a collagen gel. These data provide the first evidence that increased ZO-1 expression in melanoma contributes to the oncogenic behavior of this tumor and further illustrate that protein products of genes, such as ZO-1, can function in either a pro- or anti-oncogenic manner when expressed in different cellular contexts.

Adherens Junctions↗
PubMed2004

Hash4, a novel human achaete-scute homologue found in fetal skin.

To find additional members of the basic helix-loop-helix (bHLH) protein family in human, the bioinformatic tools provided by the National Center for Biotechnology Information was used for homology searches through databases. We report the identification and cloning of a new, fourth member in the achaete-scute complex family of genes, Hash4 or Ascl4, which maps to chromosome 12q24.1. The bHLH domain of the putative HASH4 protein shows most identity to HASH3/ASCL3. The expression of Hash4 and Hash3 was analyzed by reverse transcriptase polymerase chain reaction (PCR), in a panel of different human tissues and cells. Expression of Hash3 was detected in all samples tested, while the expression of Hash4 was restricted to skin. Quantification of the Hash4 expression by real-time PCR revealed a 7-fold higher expression in fetal skin compared to adult skin. Our results suggest HASH4 to be involved in skin development.

Amino Acid Sequence↗
PubMed2004

Connexins 26, 30, and 43: differences among spontaneous, chronic, and accelerated human wound healing.

Gap junctions (GJ) are known to be involved in spontaneous wound healing in rodent skin. We analyzed the staining patterns of the GJ proteins Cx26, Cx30, and Cx43 in human cutaneous wound healing and compared ex vivo spontaneous wound healing to non-healing wounds (chronic leg ulcers) and to ex vivo accelerated wound healing after transplantation of cultured keratinocytes. We demonstrate a loss of Cx43 staining at the wound margins during initial wound healing and after transplantation of keratinocytes. In contrast, Cx43 remains present at the margins of most non-healing wounds. We show a subsequent induction of Cx26 and Cx30 near the wound margins in spontaneous wound healing and-even earlier-after the transplantation of keratinocytes. The cells at the wound margins remain negative until the commencement of epidermal regeneration. Cx26/30 are present at the wound margins of most non-healing wounds. Cx stainings are absent in the transplanted keratinocytes during early wound healing, but there is a subsequent induction. Our results suggest that the downregulation of Cx43 is an important event in human wound healing. We discuss the assumption that direct cell-cell communication via GJ contribute to the acceleration of wound healing after the transplantation of keratinocytes.

Animals↗
PubMed2003

Expression and localization of tight junction-associated proteins in human hair follicles.

Tight junctions (TJ) are barrier-forming intercellular junctions selectively sealing cells and controlling the paracellular pathway. They have been well-characterized in simple epithelia and endothelia but have only recently been described in stratified epithelia such as epidermis, oesophagus and oral mucosa. Various epithelial layers which are partly in morphogenic continuity with the epidermis and develop therefrom during early fetal life, build the human hair follicle. The barrier function of these epithelial layers seems to be important for the universal continuity of the barrier represented by the skin. We show the presence of the TJ proteins ZO-1, occludin, and various claudins in the hair follicle and demonstrate their impressive heterogeneous distribution pattern within a given stratum as well as within its different epithelial layers. Coexpression of the various TJ proteins, arguing for typical TJ structures, can be observed especially in cell layers facing the hair shaft and the stratum corneum, and in addition at the border between the outer and inner root sheaths. Usually they are found in close proximity to desmosomal and adherens junction proteins. The morphological and biological importance of these findings and the possible roles of TJ in hair follicles, e.g. in follicular penetration, are discussed.

Adherens Junctions↗
PubMed2002

Organization and formation of the tight junction system in human epidermis and cultured keratinocytes.

Occludin and several proteins of the claudin family have been identiried in simple epithelia and in endothelia as major and structure-determining transmembrane proteins clustered in the barrier-forming tight junctions (TJ), where they are associated with a variety of TJ plaque proteins, including protein ZO-1. To examine whether TJ also occur in the squamous stratified epithelium of the interfollicular human epidermis we have applied several microscopic and biochemical techniques. Using RT-PCR techniques, we have identiried mRNAs encoding protein ZO-1, occludin and claudins 1, 4, 7, 8, 11, 12, and 17 in both tissues, skin and cultured keratinocytes, whereas claudins i and 10 have only been detected in skin tissue. By immunocytochemistry we have localized claudin-1, occludin and protein ZO-1 in distinct plasma membrane structures representing cell-cell attachment zones. While claudin-1 occurs in plasma membranes of all living cell layers, protein ZO-1 is concentrated in or even restricted to the uppermost layers, and occludin is often detected only in the stratum granulosum. Using electron microscopy, typical TJ structures ("kissing points") as well as some other apparently related junctional structures have been detected in the stratum granulosum, interspersed between desmosomes. Modes and patterns of TJ formation have also been studied in experimental model systems, e.g., during wound healing and stratification as well as in keratinocyte cultures during Ca2+-induced stratification. We conclude that the epidermis contains in the stratum granulosum a continuous zonula occludens-equivalent structure with typical TJ morphology and molecular composition, characterized by colocalization of occludin, claudins and TJ plaque proteins. In addition, cell-cell contact structures and certain TJ proteins can also be detected in other epidermal cell layers in specific cell contacts. The pattern of formation and possible functions of epidermal TJ and related structures are discussed.

Calcium↗
PubMed2002

Tight junctions and compositionally related junctional structures in mammalian stratified epithelia and cell cultures derived therefrom.

The occurrence of extended tight junction (TJ) structures, including zonulae occludentes (ZO), and the spatial arrangement of TJ proteins in stratified mammalian epithelia has long been controversially discussed. Therefore, we have systematically examined the localization of TJ proteins in diverse stratified epithelial tissues (e.g., epidermis, heel pad, snout, gingiva, tongue, esophagus, exocervix, vagina, urothelium, cornea) of various species (human, bovine, rodents) as well as in human cell culture lines derived from stratified epithelia, by electron microscopy as well as by immunocytochemistry at both the light and the electron microscopic level, using antibodies to TJ proteins such as occludin, claudins 1 and 4, protein ZO-1, cingulin and symplekin. We have found an unexpected diversity of TJ-related structures of which only those showing colocalization with the most restricted transmembrane TJ marker protein, occludin, are presented here. While in epidermis and urothelium occludin is restricted to the uppermost living cell layer, TJ-related junctions are abundant in the upper third or even in the majority of the suprabasal cell layers in other stratified epithelia. Interfollicular epidermis contains, in the stratum granulosum, extended, probably continuous ZO-like structures which can also be traced at least through the Henle cell layer of hair follicles. Similar apical ZO-like structures have been seen in the upper living cell layers of all other stratified epithelia and cell cultures examined, but in most of them we have noticed, in addition, junctional regions showing relatively broad, ribbon-like membrane contacts which in cross-section often appear pentalaminar, with an electron-dense middle lamella ("lamellated TJs", coniunctiones laminosae). In suprabasal layers of several stratified epithelia we have further observed TJ protein-containing junctions of variable sizes which are characterized by a 10-30-nm dense lamina interposed between the two membranes ("sandwich junctions"; iuncturae structae). Moreover, we have often observed variously sized regions in which the intermembrane distance is rather regularly bridged by short rod-like elements ("cross-bridged cell walls"; parietes transtillati), often in close vicinity of TJ-related structures or desmosomes. The significance of these structures and their possible biological importance are discussed.

Adult↗