PubMed Health⌕ Search

Biomedical subjects

D Garrod

Publications and source records attributed to D Garrod.

At least 19 recordsLinked to original sources
PubMed2001

Mice lacking desmocollin 1 show epidermal fragility accompanied by barrier defects and abnormal differentiation.

The desmosomal cadherin desmocollin (Dsc)1 is expressed in upper epidermis where strong adhesion is required. To investigate its role in vivo, we have genetically engineered mice with a targeted disruption in the Dsc1 gene. Soon after birth, null mice exhibit flaky skin and a striking punctate epidermal barrier defect. The epidermis is fragile, and acantholysis in the granular layer generates localized lesions, compromising skin barrier function. Neutrophils accumulate in the lesions and further degrade the tissue, causing sloughing (flaking) of lesional epidermis, but rapid wound healing prevents the formation of overt lesions. Null epidermis is hyperproliferative and overexpresses keratins 6 and 16, indicating abnormal differentiation. From 6 wk, null mice develop ulcerating lesions resembling chronic dermatitis. We speculate that ulceration occurs after acantholysis in the fragile epidermis because environmental insults are more stringent and wound healing is less rapid than in neonatal mice. This dermatitis is accompanied by localized hair loss associated with formation of utriculi and dermal cysts, denoting hair follicle degeneration. Possible resemblance of the lesions to human blistering diseases is discussed. These results show that Dsc1 is required for strong adhesion and barrier maintenance in epidermis and contributes to epidermal differentiation.

Aging↗
PubMed2001

Perinuclear and cytoplasmic distribution of desmoglein in esophageal squamous cell carcinomas.

The desmosomal glycoproteins desmoglein (Dsg) and desmocollin (Dsc) are members of the cadherin family of cell adhesion molecules. They play an important role in epithelial adhesion. To observe the distribution pattern of Dsg in esophageal squamous cell carcinomas (SCC), immunohistochemical and immunoelectron microscopic analyses were performed. Immunohistochemically, normal esophageal squamous cells strongly expressed Dsg at the cell-cell boundaries, while moderately differentiated esophageal SCC cells showed a perinuclear distribution in addition to the cell boundary staining. At the ultrastructural level, the reaction product was concentrated at the desmosomes in the cell membrane region of normal epithelial cells, but was reduced at the membrane and found throughout the cytoplasm as well as in the surrounding outer nuclear envelope in SCC cells. These results demonstrate an aberrant distribution of Dsg in SCC cells. This may have important consequences for invasion and metastasis, as it may indicate loosened intercellular adhesion.

Aged↗
PubMed2000

Primary care groups. Fighting chance.

Interviews with PCG chief executives, six months into their job, revealed concerns about the personal qualities needed for the role. There were concerns about the dominance of GPs on boards, at the expense of nurse members. Some reported that health authorities had difficulty in letting go of control to the new organisations.

Decision Making, Organizational↗
PubMed2000

The alpha isoform of protein kinase C is involved in signaling the response of desmosomes to wounding in cultured epithelial cells.

Initiation of reepithelialization upon wounding is still poorly understood. To enhance this understanding, we focus here on changes in the adhesive state of desmosomes of cultured Madin-Darby canine kidney cells in response to wounding of confluent cell sheets. Previous results show that desmosomal adhesion in Madin-Darby canine kidney cells changes from a calcium-dependent state to calcium independence in confluent cell sheets. We show that this change, which requires culture confluence to develop, is rapidly reversed upon wounding of confluent cell sheets. Moreover, the change to calcium dependence in wound edge cells is propagated to cells hundreds of micrometers away from the wound edge. Rapid transition from calcium independence to calcium dependence also occurs when cells are treated with phorbol esters that activate PKC. PKC inhibitors, including the conventional isoform inhibitor Gö6976, cause rapid transition from calcium dependence to calcium independence, even in subconfluent cells. The cellular location of the alpha isoform of PKC correlates with the calcium dependence of desmosomes. Upon monolayer wounding, PKCalpha translocates rapidly to the cell periphery, becomes Triton X-100 insoluble, and also becomes concentrated in lamellipodia. The PKCalpha translocation upon wounding precedes both the increase in PKC activity in the membrane fraction and the reversion of desmosomes to calcium dependence. Specific depletion of PKCalpha with an antisense oligonucleotide increases the number of cells with calcium-independent desmosomes. These results show that PKCalpha participates in a novel signaling pathway that modulates desmosomal adhesion in response to wounding.

Animals↗
PubMed1998

Desmosomal adhesion inhibits invasive behavior.

Recent studies of human disease and transgenic animal experiments have clearly demonstrated the importance of desmosomes in normal tissue architecture. Furthermore, desmosomal components are down-regulated in certain types of carcinomas, suggesting a possible role for desmosomes in suppression of invasion and metastasis. However, there is no functional evidence to support such a hypothesis. To obtain such evidence, we needed to generate desmosomal adhesion in an invasive cell line. We show that expression of multiple desmosomal components (the desmosomal cadherins, desmocollin and desmoglein, and the armadillo protein, plakoglobin) in nonadhesive L929 fibroblasts generates adhesion in aggregation assays. This adhesion is specifically blocked by short peptides corresponding to the putative cell adhesion recognition sites of desmocollin and desmoglein. This result provides an experimental demonstration of the functional importance of the cell adhesion recognition sites of desmocollin and desmoglein and indicates that both desmosomal cadherins are specifically involved in this adhesion. Moreover, whereas parental L929 cells are strongly invasive into collagen gels, we show that invasion is substantially inhibited in cells transfected with desmosomal components. Invasion is restored by treating the transfected cells with anti-adhesion peptides, indicating that desmosomal adhesion specifically blocks invasion in culture. Our results support the suggestion that desmosomes have a role in suppression of tumor spreading.

Cadherins↗
PubMed1996

Desmosomes: differentiation, development, dynamics and disease.

Recent evidence on the distribution of desmosomal glycoprotein isoforms that shows their combined expression in individual desmosomes has strengthened the belief that the latter are involved in epithelial differentiation and morphogenesis. It has been shown that cellular interactions and protein kinase C can modulate the adhesive properties of desmosomes in epithelial cell sheets. Genetic studies indicate the involvement of desmosomal components in cancer and epidermal diseases.

Animals↗
PubMed1995

Sulphated proteoglycan is required for collecting duct growth and branching but not nephron formation during kidney development.

Kidney epithelia have separate origins; collecting ducts develop by ureteric bud growth and arborisation, nephrons by induced mesenchyme-epithelium transition. Both express sulphated glycosaminoglycans (GAGs) which are strikingly upregulated during nephron differentiation. However, sodium chlorate, an inhibitor of GAG sulphation, and the GAG-degrading enzymes heparitinase plus chondroitinase, did not prevent nephron development. In contrast, ureteric bud growth and branching were reversibly inhibited by the above reagents, the inhibition correlating quantitatively with sulphated GAG deprivation caused by a range of chlorate concentrations. Growth and branching could be independently restored during GAG deprivation by hepatocyte growth factor and phorbol-12-myristate acetate (PMA) respectively. Together these signalling effectors stimulated both branch initiation and growth. Thus growth and morphogenesis of ureteric bud involve distinct signalling pathways both regulated by GAGs.

Animals↗
PubMed1995

A simple epithelial cell line (MDCK) shows heterogeneity of desmoglein isoforms, one resembling pemphigus vulgaris antigen.

The epidermal blistering disease, pemphigus vulgaris (PV), is caused by circulating autoantibodies that react with a desmosomal glycoprotein desmoglein (Dsg3). This antigen is expressed only in stratified epithelial tissues. Here we show that the simple epithelial canine kidney cell line, MDCK, expresses at least two desmoglein isoforms recognised by different monoclonal antibodies. One of these isoforms is a 130 x 10(3) M(r) polypeptide that is recognised by both PV autoantisera and a monoclonal antibody reactive with a cytoplasmic domain of human Dsg3. Antibodies in PV sera bind to the surface of MDCK cells but not cause loss of intercellular adhesion. This is the first demonstration of the expression of a polypeptide related to human PV antigen by a simple epithelial cell type.

Animals↗
PubMed1993

Epithelial cells retain junctions during mitosis.

It has long been known that cells show reduced cell-substratum adhesion during mitosis in tissue culture, but it is not generally known whether cell-cell adhesion is also reduced. Epithelial cells, both in culture and in tissues, are linked together by several different types of intercellular junctions. Are these junctions disassembled when epithelial cells divide? Cultured epithelial cells were fluorescently stained for desmosomes, tight junctions and zonulae adherentes, and large numbers of dividing cells examined by light microscopy. The results suggested that all three types of intercellular junctions were retained throughout cell division and no evidence for internalization of junctions was obtained. The persistence of intercellular junctions by cultured cells during division was confirmed by electron microscopy. In order to determine whether intercellular junctions were similarly retained by dividing cells in tissues, human colonic mucosal crypt cells and basal keratinocytes were studied by electron microscopy. Both cell types retained intercellular junctions during division. Dividing basal keratinocytes also possessed hemidesmosomal contact with the basement membrane. It is suggested that retention of cellular junctions during division is important for maintenance of tissue integrity and organization.

Actins↗
PubMed1993

Confocal and conventional immunofluorescence and ultrastructural localisation of intracellular strength-giving components of human amniochorion.

Key cytoskeletal polypeptides of human fetal membranes have been localised at subcellular level using confocal and conventional indirect immunofluorescence microscopy. Correlation with electron microscope data has allowed us to examine how cellular compartments of this multilaminar tissue maintain their mechanical integrity until the time of membrane rupture at parturition. Evidence is presented for myofibroblastic characteristics of cells in both the fibroblast and reticular layers which may therefore have tension-generating, position-adjustment and wound-healing roles in the amniochorion. Desmin and vimentin are coexpressed in these cells, but a small localised population of cells in the fibroblast layer contains vimentin alone. An interaction of cytokeratin filaments with nuclei and desmosomes of amniotic epithelium in vivo is demonstrated, indicating that nuclei of cells of ectodermal origin are integrated into a mechanical structure extending throughout the tissue as a whole. Cells of the basal 1 or 2 layers of trophoblast have been shown to have a more extensive and better integrated cytoskeletal organisation than those overlying and forming the boundary with decidua. Structures within the trophoblast, identified previously as degenerate villi, contain cells with intermediate filaments with similar immunofluorescence properties to those of the neighbouring reticular layer and thus may represent papillae that prevent shearing at this interface.

Amnion↗
PubMed1987

Localisation of the protein and glycoprotein components of bovine nasal epithelial desmosomes by immunoelectron microscopy.

Desmosomal proteins (dp1-4) and glycoproteins (dg1-3) have been localised within desmosomes of bovine nasal epithelium by immunogold labelling of ultrathin frozen sections. Beginning in the extracellular space and proceeding through the plaque to the tonofilaments, the following localisations were found. Labelling for the 130,000 and 115,000 Mr glycoproteins (dg2 and dg3) was predominantly in the extracellular space, a location consistent with their proposed adhesive function. The glycoproteins of 175,000-164,000 Mr (dg1) were also found in the extracellular space and in addition had cytoplasmic domains extending throughout the cytoplasmic plaque. The 83,000 Mr protein (dp3) was located along the cytoplasmic face of the membrane and extended into the plaque, whereas an antibody which recognises both the 83,000 Mr protein (dp3) and the 75,000 Mr protein (dp4) gave labelling both in and beyond the plaque. Labelling for the high mol. wt proteins of Mr 250,000 and 215,000 (dp1 and dp2) was largely excluded from the plaque, and was located distally, adjacent to the tonofilaments. Hemidesmosomes could not be labelled with antibodies to dg1-3 or dp3 and 4, but some labelling was obtained with antibody to dp1 and 2.

Animals↗
PubMed1986

An investigation of the molecular components of desmosomes in epithelial cells of five vertebrates.

We have shown previously, by fluorescent antibody staining, that desmosomal antigens are widely distributed in the tissues of vertebrate animals. Furthermore, we have demonstrated mutual desmosome formation between cells derived from man, cow, dog, chicken and frog. In this paper we have studied the components of desmosomes in a tissue or a cell line from each of these animals by immunoblotting with antibodies raised against the desmosomal components isolated from bovine nasal epithelium. Blotting was carried out on bovine nasal epithelial desmosomal cores, desmosome-enriched fractions derived from chicken and frog epidermis, nuclear matrix-intermediate filament scaffolds derived from Madin-Darby bovine and canine cells (MDBK and MDCK), and unextracted cultured human foreskin keratinocytes. The results show that desmosomes from all these sources contain high molecular weight proteins (desmoplakins) of similar or identical molecular weights (250 000 and 215 000). Antibodies against the two lower molecular weight desmosomal proteins (83 000 and 75 000) always recognized one or two bands in very similar molecular weight regions of the gels. The desmosomal glycoproteins were found to be much more variable than the proteins: they vary between sources in molecular weight, heterogeneity and antibody cross-reactivity. For instance, antibody specific for a group of glycoprotein bands of 175 000, 169 000 and 164 000 (Mr) in bovine nasal epithelium recognizes three bands of 245 000, 230 000 and 210 000 in MDCK cells but only a single band of 190 000 in keratinocytes. In mammals, the 175 000-164 000 glycoproteins and the desmosomal adhesion molecules, the desmocollins (Mr 130 000 and 115 000 in cow's nose), are immunologically distinct. In chicken and frog, however, there are glycoproteins that react with both anti-175 000-164 000 and anti-desmocollin antibodies, but there are also distinct desmocollin bands. The significance of these results is discussed in relation to conservation of desmosomal components and adhesion mechanisms. It is suggested that adhesion may be performed by a well-conserved protein domain and that the variation between desmosomal glycoproteins from different sources may be due to differences in their carbohydrate composition.

Animals↗
PubMed1984

Distribution of desmosomal components in the tissues of vertebrates, studied by fluorescent antibody staining.

In previous work we used immunofluorescent staining with specific antibodies to study the distribution of five desmosomal antigens in the epithelia of different vertebrate animals. We showed that all five antigens were present in all epithelia studied in human, bovine, rat, guinea pig, chick and frog (Rana pipiens) tissues. It was concluded that desmosomes are highly conserved structures. This paper extends those studies: by including three other species, a lizard (Lacerta viridis), the axolotl (Ambystoma mexicanum) and the trout (Salmo trutta), and by looking at several tissues in more detail. The principal results are as follows. The epidermis of all species down to the frog stain with equal intensity for all desmosomal antigens. In the epidermis of axolotl and trout, staining for desmosomal plaque constituents is present, but staining for the desmosomal glycoproteins is greatly reduced or absent. Within mammalian species as well as chick, lizard and frog, staining for the 115 X 10(3) and 100 X 10(3) molecular weight desmosomal glycoproteins is less intense in non-epidermal tissues than in the epidermis, while staining for desmosomal plaque constituents and for the 150 X 10(3) molecular weight glycoprotein is undiminished. It is possible, therefore, that slight differences exist between certain glycoproteins of epidermis and non-epidermal epithelia. The hearts of lower vertebrates (lizard, frog, axolotl and trout) stain only for individual desmosomal plaque antigens. The pillar cells of trout gill stain, adjacent to their collagenous columns, for one desmosomal plaque antigen. There is a fibrous cytoplasmic mat in this position but no desmosomes. Thus one of the desmosomal antigens may have a function outside the desmosome.

Ambystoma mexicanum↗
PubMed1984

HeLa cells form focal contacts that are not fibronectin dependent.

HeLa cells cultured on glass substrata produce numerous prominent focal contacts, which reside at the termini of actin microfilament bundles. However, very few of the cells stain for fibronectin with specific anti-fibronectin antibody. Moreover, the cells form focal contacts in fibronectin-depleted medium, in the presence of high concentrations of anti-fibronectin immunoglobulin G and in the presence of monensin. No fibronectin synthesis can be detected by [35S]methionine-labelling and immunoprecipitation. The possibility that HeLa cell focal contacts are independent of fibronectin in their formation is discussed in relation to the controversy about the relationship between fibronectin and focal contacts.

Animals↗
PubMed1984

Identification of desmosomal surface components (desmocollins) and inhibition of desmosome formation by specific Fab'.

Specific antibodies against the components of desmosomes, the adhesive junctions of epithelial cells, have been used to determine which components are located on the cell surface. Three criteria have been used: fluorescent antibody staining, immuno-gold labelling and electron microscopy, and quantitative measurements of antibody binding using [125I]protein A. When these techniques were applied to living Madin-Darby bovine kidney (MDBK) cells, antibodies against only two desmosomal components, glycoproteins of approximately 115 X 10(3) Mr and 100 X 10(3) Mr, bound to the cell surface. Antibodies against all other components, the 230 and 205 X 10(3) Mr proteins (desmoplakins), the 150 X 10(3) Mr glycoprotein and the 82 and 86 X 10(3) Mr proteins reacted in fluorescent antibody staining only after cells had been fixed and made permeable. MDBK cells were cultured in the presence of univalent fragments (Fab') of anti-desmosomal antibodies for periods from 24 h to 72 h. After these times cells were fixed, made permeable, and stained with anti-desmoplakin antibody to assay for desmosome formation. Fab' derived from anti-100 X 10(3) Mr protein specifically inhibited desmosome formation, whereas Fab's from anti-desmoplakin, anti-150 X 10(3) Mr and anti-82 and 86 X 10(3) Mr proteins were without effect. We conclude that the 100 X 10(3) Mr and the immunologically related 115 X 10(3) Mr components are located on the cell surface and are directly involved in cell-cell adhesion. We have named them desmocollins to denote that they are involved in the adhesive function of desmosomes. The modulation of desmocollin distribution during monolayer formation and establishment of epithelial polarity has also been studied. Fluorescent and immuno-gold labelling using Fab' or IgG at 4 degrees C revealed that desmocollins were initially evenly dispersed over the cell surface. Staining with IgG at 37 degrees C caused the desmocollins to "patch' but not to "cap'. With the establishment of confluency, desmocollins were gradually removed from the upper surfaces of the cells (or masked and rendered inaccessible to antibody) being confined to the lateral and probably basal regions of the cells. Treatment of confluent monolayers with 3 mM-EGTA rendered the desmocollins stainable, probably by causing their release from lateral constraint. Desmocollin staining at the cell surface was not appreciably reduced during 5 h of EGTA treatment, suggesting that desmocollins, unlike desmosomal plaques, may not be internalized after junction breakdown.

Animals↗
PubMed1982

Effects of vitamin A on the behaviour of migratory neural crest cells in vitro.

It has been proposed elsewhere that the teratogenic effects of retinoids on craniofacial morphogenesis are caused by a disturbance of the migration of cranial neural crest cells. The effects of 3.5 X 10(-5) M and 3.5 X 10(-6) M-retinol on the migration of avian neural crest cells in vitro have been investigated by monitoring cell morphology, locomotory behaviour, fibronectin distribution and actin-microfilament organization. Retinol retards migration by affecting cell-to-substratum adhesiveness. Cells exposed to medium containing retinol are less adherent to the substratum, and although the cell surface is very mobile, are unable to extend or maintain lamellipodia. As a consequence the cells do not actively translocate. Fibronectin distribution at the cell surface is sparse, possibly as a result of shedding, and actin distribution remains diffuse. At the retinol molarities used all these effects are reversible. Thus cells allowed to recover in normal medium flatten out, display lamellipodia and commence active translocation. Fibronectin becomes organized into a fibrillar array and actin microfilaments become organized into cables. The period needed for this recovery is directly related to the molarity of retinol during the initial exposure; after recovery the retinol-treated cells are virtually indistinguishable from control cells. We propose that in vivo the effects of retinoids might be to impair cell-extracellular matrix interaction, thus impeding a cell's ability to migrate through that matrix. Contrary to previous suggestions, the in vivo effects are probably not in any way 'specific' to neural crest cells but are more accurately considered as 'selective', in that any cell undergoing migration would be similarly affected.

Actins↗