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Biomedical subjects

B C Powell

Publications and source records attributed to B C Powell.

At least 19 recordsLinked to original sources
PubMed2004

Structure and expression of the ovine Hoxc-13 gene.

HOXC-13 has an important role in controlling hair formation through regulating keratin differentiation-specific genes. In this study, we describe the isolation and characterisation of the Hoxc-13 gene from sheep wool follicles and its expression in the skin. We show that the gene organisation of ovine Hoxc-13 is similar to other homeobox genes of the Abd-B type I Homeobox class with two exons split by an intron next to the homeobox. The gene spans 7.5 kilobases (kb) and has a relatively large intron, which divides an open reading frame of 2361 nucleotides. The predicted ovine Hoxc-13 protein of 330 amino acids has over 97% sequence identity with the human and mouse proteins. A second novel transcript was identified, which could produce a truncated Hoxc-13 protein lacking 15 amino acids from the N-terminus. A positionally conserved Hoxc-13 binding site in the Hoxc-13 proximal promoters of sheep, human, mouse and newt suggests that Hoxc-13 expression is autoregulatory. Positionally conserved motifs for LEF-1 and Whn/Foxn1 suggest that Hoxc-13 may be a downstream target of these transcription factors known to regulate hair growth. In addition to expression in the follicle, we detected Hoxc-13 in cells of the blood sinus surrounding vibrissal follicles and in scattered cells in the upper dermis of the skin. Thus, in addition to a role in controlling transcription of hair keratins, Hoxc-13 may have other roles in skin function.

5' Flanking Region↗
PubMed2003

Maternal milk regulation of cell infiltration and interleukin 18 in the intestine of suckling rat pups.

BACKGROUND: and aims: In neonates the gastrointestinal tract is exposed to food and bacterial antigens at a time when the gut mucosal immune system has not developed the ability to induce oral tolerance. This increases the risk for an inappropriate immune response to oral antigens. Transforming growth factor beta (TGF-beta) is an immunoregulatory cytokine present in high concentration in maternal milk. Interleukin 18 (IL-18) is a cytokine that mediates early immune events, and drives T cell development. We assessed the role of TGF-beta in mediating mucosal immune development and specifically the effect on endogenous IL-18. METHODS: Rat pups were randomly assigned to the following groups, naturally suckled, maternal milk via cannula, and formula fed with and without physiological levels of TGF-beta2. A comparison of the immune response profile was then carried out. Cytokine profiles, dendritic cell, intestinal mast cell, and eosinophil numbers were assessed. RESULTS: We show that feeding formula deficient in TGF-beta2 resulted in accumulated IL-18 protein release from intestinal epithelial cells and IL-18 mRNA up regulation. A proinflammatory cytokine profile resulted in the gut, along with increased numbers of activated dendritic cells, eosinophils, and mast cells. Supplementation of the formula with TGF-beta2 down regulated the proinflammatory cytokine mRNA as well as the number of activated lymphocytes, eosinophils, mast cells, CD80, and CD86 positive dendritic cells. CONCLUSION: The data suggests an important role for maternal milk, in regulating immune responses after exposure to food antigens, which might otherwise induce deleterious immune responses in the intestine of suckling neonates. This regulation is potentially mediated by milk TGF-beta2, as well as endogenous IL-18.

Animals↗
PubMed2001

A millennial myosin census.

The past decade has seen a remarkable explosion in our knowledge of the size and diversity of the myosin superfamily. Since these actin-based motors are candidates to provide the molecular basis for many cellular movements, it is essential that motility researchers be aware of the complete set of myosins in a given organism. The availability of cDNA and/or draft genomic sequences from humans, Drosophila melanogaster, Caenorhabditis elegans, Arabidopsis thaliana, Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Dictyostelium discoideum has allowed us to tentatively define and compare the sets of myosin genes in these organisms. This analysis has also led to the identification of several putative myosin genes that may be of general interest. In humans, for example, we find a total of 40 known or predicted myosin genes including two new myosins-I, three new class II (conventional) myosins, a second member of the class III/ninaC myosins, a gene similar to the class XV deafness myosin, and a novel myosin sharing at most 33% identity with other members of the superfamily. These myosins are in addition to the recently discovered class XVI myosin with N-terminal ankyrin repeats and two human genes with similarity to the class XVIII PDZ-myosin from mouse. We briefly describe these newly recognized myosins and extend our previous phylogenetic analysis of the myosin superfamily to include a comparison of the complete or nearly complete inventories of myosin genes from several experimentally important organisms.

Animals↗
PubMed2000

Expression of the homeobox gene, Barx2, in wool follicle development.

We have cloned ovine Barx2, a member of the Bar class of homeobox genes, and present the first description of Barx2 expression in wool follicle development. Barx2 is uniformly expressed in the embryonic ectoderm but is transiently downregulated during the initiation of follicle morphogenesis. Subsequently, Barx2 is expressed throughout the epithelial component of the developing follicle except for a small group of cells at the leading edge of the follicle placode. These Barx2-negative cells are destined to form the follicle bulb and are the progenitors of the inner root sheath and hair shaft. In adult follicles, Barx2 is expressed throughout the outer root sheath but not in the inner root sheath or hair shaft, or in dermal cells associated with the follicle. The pattern of Barx2 expression in follicle morphogenesis is similar to that of the cell adhesion molecule E-cadherin, a similarity that echoes Barx2 coexpression with the L1 cell adhesion molecule in other tissues during mouse embryogenesis. Barx2 is also expressed in tongue and esophagus, two other keratinizing tissues, and we speculate that Barx2 may have a general function in controlling adhesive processes in keratinizing epithelia.

Animals↗
PubMed2000

Transsulfuration, protein synthesis rate and follicle mRNA in the skin of young Merino lambs in response to infusions of methionine and serine.

Methionine (Met) is usually the first limiting amino acid for sheep and supplements of Met may increase production of wool and meat. The wool response may be due to an increased supply of cysteine (Cys) from transsulfuration (TS) of Met. Met is catabolized through homocysteine to form Cys when the S from Met is transferred to serine (Ser). We hypothesized that providing additional Met would create a deficiency of Ser and that by simultaneously providing Met and Ser, TS and wool growth could be increased more than by providing Met alone. The effects of i.v. infusions of Met and Ser to young Merino lambs on TS, fractional synthesis rate (FSR) of protein in skin, follicle mRNA and wool growth were examined. Following 4 d of constant i.v. infusion of 3 g Met/d, or 10 g Ser/d or both, the isotope tracers: L-[3-(13)C]Cys, L-[ring-d5]phenylalanine (Phe) and L-[2,3,3-d3]Ser were infused over 8 h to allow for measurements of irreversible loss rate (ILR), and TS in whole body and skin. Skin biopsies were taken for measurement of FSR. Wool growth rate was measured using autoradiography. An infusion of Met significantly (P < 0.05) improved wool growth rate and increased skin FSR, Cys supply from TS and enhanced levels of follicle mRNA (from the K2.10 intermediate filament gene and three gene families encoding keratin associated proteins KAP1, KAP4 and KAP12). The extra Met lowered Ser ILR. The infusion of Ser doubled Ser ILR in the body and increased skin FSR calculated using the Cys tracer in plasma (P < 0.05). However, there were no significant (P > 0.05) changes in TS, skin FSR calculated using the Phe and Ser tracers, follicle mRNA or wool growth rate as a result of Ser infusion. While there were trends towards increased TS and FSR with Ser infusion, the overall lack of significant changes indicates a high capacity for the de novo synthesis of Ser.

Animals↗
PubMed1999

Dynamic expression of ornithine decarboxylase in hair growth.

Ornithine decarboxylase (ODC) is the key enzyme in the synthesis of polyamines, small cationic molecules believed to have a role in many cellular processes such as cell migration, proliferation and differentiation. We show that ODC expression is associated with cell proliferation and commitment in hair follicle development and hair growth. In embryonic epidermis, ODC is expressed in ectodermal cells at sites where follicles develop, and persists in cells at the leading edge of the follicle placode. ODC is abundantly expressed in proliferating bulb cells of anagen follicles, except for a pocket of cells at the base of the bulb. Entry of the follicle into catagen is accompanied by a down-regulation of ODC expression, which is not resumed until a new follicle is initiated. In vibrissae, ODC expression is more complex. ODC is expressed not only in the bulb but also in the hair shaft, presenting a striking biphasic pattern. Additionally, ODC is expressed in a group of outer root sheath cells in the vicinity of the follicle bulge, the putative site of hair follicle stem cells.

Animals↗
PubMed1998

Expression of the intermediate filament keratin gene, K15, in the basal cell layers of epithelia and the hair follicle.

The intermediate filament keratin, K15, is present in variable abundance in stratified epithelia. In this study we have isolated and characterized the sheep K15 gene, focusing on its expression in the follicles of sheep and mice. We show that K15 is expressed throughout the hair cycle in the basal layer of the outer root sheath that envelops the follicle. Strikingly, however, in large medullated wool follicles, a small group of basal outer root sheath cells located in the region thought to contain hair follicle stem cells are K15-negative. In the follicle bulb K15 is expressed in cells situated next to the dermal papilla but not in the inner bulb cells. Elsewhere, K15 is expressed at a low, variable level in the basal layer of the epidermis and sebaceous gland, often in a punctate pattern. In the esophagus of the sheep K15 expression is restricted to the basal layer, in contrast to human esophagus where it is expressed throughout the epithelium. Transgenic mouse lines established with a 15-kb sheep K15 gene construct exhibited faithful expression and showed no phenotypic consequences of K15 overexpression. An investigation of transgene expression showed that K15 is continuously expressed in outer root sheath cells during the hair cycle. Given its expression in the mitotically active basal cell layers of diverse epithelia and the follicle, K15 expression appears to signal an early stage in the pathway of keratinocyte differentiation that precedes the decision of a cell to become epidermal or hair-like.

Amino Acid Sequence↗
PubMed1998

The Notch signalling pathway in hair growth.

The Notch signalling pathway is an important mediator of cell fate selection whose involvement in epidermal appendage formation is now becoming recognised. Hair follicle development and hair formation involve the co-ordinated differentiation of several different cell types in which Notch appears to have a role. We report intricate expression patterns for the Notch-1 receptor and three ligands, Delta-1, Jagged-1 and Jagged-2 in the hair follicle. Notch-1 is expressed in ectodermal-derived cells of the follicle, in the inner cells of the embryonic placode and the follicle bulb, and in the suprabasal cells of the mature outer root sheath. Delta-1 is only expressed during embryonic follicle development and is exclusive to the mesenchymal cells of the pre-papilla located beneath the follicle placode. Expression of Jagged-1 or Jagged-2 overlaps Notch-1 expression at all stages. In mature follicles, Jagged-1 and Jagged-2 are expressed in complementary patterns in the follicle bulb and outer root sheath, Jagged-1 in suprabasal cells and Jagged-2 predominantly in basal cells. In the follicle bulb, Jagged-2 is localised to the inner (basal) bulb cells next to the dermal papilla which do not express Notch-1, whereas Jagged-1 expression in the upper follicle bulb overlaps Notch-1 expression and correlates with bulb cell differentiation into hair shaft cortical and cuticle keratinocytes.

Animals↗
PubMed1998

Expression of a wool intermediate filament keratin transgene in sheep fibre alters structure.

Alteration of the protein composition of the wool fibre via transgenesis with sheep wool keratin and keratin associated protein (KAP) genes may lead to production of fibre types with improved processing and wearing qualities. Using this approach, we have demonstrated that high level cortical-specific expression of a wool type II intermediate filament (IF) keratin gene, K2.10, leads to marked alterations in both the microstructure and macrostructure of the wool fibres, which have higher lustre and reduced crimp. Analysis of mRNA found reduced levels of transcripts from endogenous cortical type I (p < 0.05) and type II (p < 0.01) keratin IF genes and from the KAP8 (p < 0.001) and KAP2 (p < 0.01) gene families. Examination of protein composition revealed an altered ratio in the keratin type II protein family of the wool fibre cortex. Whilst the over-expressed K2.10 transgene product constituted the majority of keratin type II IF protein, it appeared unable to form heterodimers with much of the expressed endogenous keratin type I IF. In comparison with non-transgenic sheep, fewer IF microfibrils were visible in the cortical cells of fibres from transgenics. The combined effect on fibre structure was disruption of the formation of orthocortical and paracortical cells in the fibre cortex, a factor which could account for the reduction in fibre crimp. No effects upon transcript or protein levels, or fibre microstructure or macrostructure were observed in transgenic sheep expressing the transgene at lower levels, indicating that subtle changes to the gene expression profile in sheep wool follicles can be tolerated. The data here also illustrate that control over endogenous transcript levels in the cortex results when factors acting on the endogenous keratin type I, keratin type II and KAP gene sequences are sequestered by the active K2.10 transgene locus. Moreover, interference to a transcriptional hierarchy shared by keratin and KAP genes may occur prior to establishment of the orthocortical and paracortical compartments of the follicle cortex, at the level of the chromatin.

Animals↗
PubMed1998

Regulation of a hair follicle keratin intermediate filament gene promoter.

During hair growth, cortical cells emerging from the proliferative follicle bulb rapidly undergo a differentiation program and synthesise large amounts of hair keratin proteins. To identify some of the controls that specify expression of hair genes we have defined the minimal promoter of the wool keratin intermediate filament gene K2.10. The region of this gene spanning nucleotides -350 to +53 was sufficient to direct expression of the lacZ gene to the follicle cortex of transgenic mice but deletion of nucleotides -350 to -150 led to a complete loss of promoter activity. When a four base substitution mutation was introduced into the minimal functional promoter at the binding site for lymphoid enhancer factor 1 (LEF-1), promoter activity in transgenic mice was decreased but specificity was not affected. To investigate the interaction of trans-acting factors within the minimal K2.10 promoter we performed DNase I footprinting analyses and electrophoretic mobility shift assays. In addition to LEF-1, Sp1, AP2-like and NF1-like proteins bound to the promoter. The Sp1 and AP2-like proteins bound sequences flanking the LEF-1 binding site whereas the NF1-like proteins bound closer to the transcription start site. We conclude that the LEF-1 binding site is an enhancer element of the K2.10 promoter in the hair follicle cortex and that factors other than LEF-1 regulate promoter tissue- and differentiation-specificity.

Animals↗
PubMed1997

The role of keratin proteins and their genes in the growth, structure and properties of hair.

The importance of wool in the textile industry has inspired extensive research into its structure since the 1960s. Over the past several years, however, the hair follicle has increased in significance as a system for studying developmental events and the process of terminal differentiation. The present chapter seeks to integrate the expanding literature and present a broad picture of what we know of the structure and formation of hair at the cellular and molecular level. We describe in detail the hair keratin proteins and their genes, their structure, function and regulation in the hair follicle, and also the major proteins and genes of the inner and outer root sheaths. We discuss hair follicle development with an emphasis on the factors involved and describe some hair genetic diseases and transgenic and gene knockout models because, in some cases, they stimulate natural mutations that are advancing our understanding of cellular interactions in the formation of hair.

Amino Acid Sequence↗
PubMed1995

Characterization of a gene encoding a cysteine-rich keratin associated protein synthesized late in rabbit hair follicle differentiation.

Many different cysteine-rich proteins are synthesized during hair follicle differentiation, forming part of the interstitial matrix between bundles of intermediate filaments. We have isolated a rabbit gene (rKAP4L), a member of a multigene family that encodes a small cysteine-rich hair keratin associated protein. This is the first complete gene sequence for this family. The rKAP4L gene is expressed in the cortex of rabbit pelage hair follicles at a late stage of hair follicle differentiation, well after the synthesis of the other major hair proteins, the intermediate filament and glycine/tyrosine-rich keratin associated proteins, has commenced. The protein contains 36 mol % cysteine, with a molecular size of 13593 daltons, and its sequence appears to be based on a pentapeptide repeat. It is predicted to adopt a folded conformation characterized by beta-turns interspersed with short stretches of beta-sheet or random coil.

Amino Acid Sequence↗
PubMed1994

Transgenic sheep and wool growth: possibilities and current status.

Merino wool is the result of generations of selection, yet improvements in wool quality and performance are still being sought. Through gene manipulation, sheep transgenesis offers possibilities of understanding the relationship between wool keratin protein composition and fibre structure and properties and of introducing novel changes to fibre properties and growth rates. We have established an efficient sheep transgenesis programme with an overall transgenic rate of 2.1% of zygotes injected. However, by incorporating in vitro culture and assessment of injected zygotes, this equates to a transgenic rate of 13% from 516 lambs born. With the first keratin gene construct, a wool keratin type II intermediate filament gene, four live F0 transgenic sheep have been produced and all express the transgene. In one of them, the highest expressor, phenotypic and ultrastructural changes were evident in the fleece. To improve wool growth rate by increasing the supply of cysteine to the follicle, transgenic sheep are being produced carrying the two genes necessary for endogenous cysteine synthesis. Three promoters have been tested driving the cysteine synthesis genes: two general promoters, the Rous sarcoma virus long terminal repeat and mouse phosphoglycerate kinase promoter, and a rumen-specific promoter from the sheep small proline-rich protein gene. To date, one transgenic sheep (bearing the small proline-rich protein promoter constructs) has produced cysteine in the rumen, although the amount was low at 3 months of age and not detectable at 6 months.

Animals↗