scholarly journals Impaired Hair Follicle Morphogenesis and Polarized Keratinocyte Movement upon Conditional Inactivation of Integrin-linked Kinase in the Epidermis

2008 ◽  
Vol 19 (4) ◽  
pp. 1462-1473 ◽  
Author(s):  
Kerry-Ann Nakrieko ◽  
Ian Welch ◽  
Holly Dupuis ◽  
Dawn Bryce ◽  
Agnieszka Pajak ◽  
...  
Keyword(s):  
Hair Follicle ◽  
Developmental Stages ◽  
Single Cells ◽  
Forward Movement ◽  
Keratinocyte Proliferation ◽  
Cultured Keratinocytes ◽  
Conditional Inactivation ◽  
Integrin Linked Kinase ◽  
Key Aspects

Integrin-linked kinase (ILK) is key for cell survival, migration, and adhesion, but little is known about its role in epidermal development and homeostasis in vivo. We generated mice with conditional inactivation of the Ilk gene in squamous epithelia. These mice die perinatally and exhibit skin blistering and severe defects in hair follicle morphogenesis, including greatly reduced follicle numbers, failure to progress beyond very early developmental stages, and pronounced defects in follicular keratinocyte proliferation. ILK-deficient epidermis shows abnormalities in adhesion to the basement membrane and in differentiation. ILK-deficient cultured keratinocytes fail to attach and spread efficiently and exhibit multiple abnormalities in actin cytoskeletal organization. Ilk gene inactivation in cultured keratinocytes causes impaired ability to form stable lamellipodia, to directionally migrate, and to polarize. These defects are accompanied by abnormal distribution of active Cdc42 to cell protrusions, as well as reduced activation of Rac1 upon induction of cell migration in scraped keratinocyte monolayers. Significantly, alterations in cell spreading and forward movement in single cells can be rescued by expression of constitutively active Rac1 or RhoG. Our studies underscore a central and distinct role for ILK in hair follicle development and in polarized cell movements, two key aspects of epithelial morphogenesis and function.

scholarly journals Dynamics of the transcriptome during chicken embryo development based on primordial germ cells

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Aleksandra Dunislawska ◽  
Agata Szczerba ◽  
Maria Siwek ◽  
Marek Bednarczyk
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Germ Cells ◽  
Developmental Stages ◽  
Single Cells ◽  
Primordial Germ Cells ◽  
Regulation Of Gene Expression ◽  
Cell Number ◽  
Early Developmental

Abstract Objective Regulation of gene expression during embryo development on the basis of migration of primordial germ cells (PGCs) in vivo has been rarely studied due to limited cell number and the necessity to isolate PGCs from a large number of embryos. Moreover, little is known about the comprehensive dynamics of the transcriptome in chicken PGCs during early developmental stages. The current study investigated transcriptome dynamics of chicken PGCs at key developmental stages: 4.5, 8 and 12 days of embryo incubation. PGCs were collected, and RNA was isolated using a commercial kit for single cells. The isolated RNA was subjected to microarray analysis (Agilent Technologies). Results Between 8 and 12 days of incubation, the highest number of genes was regulated. These data indicate that the most intense biological activity occurs between 8 and 12 days of embryo development. Heat map showed a significant decrease in gene expression on day 8, while it increased on day 12. The development of a precise method to isolate bird PGCs as well as the method to isolate RNA from single cells isolated from one embryo allows for early molecular analysis and detection of transcriptome changes during embryonic development.

Cell Morphologies and Contacts in Annual Fish Embryos

1980 ◽  
Vol 38 ◽  
pp. 706-707
Author(s):  
Roland Lesseps
Keyword(s):  
Developmental Stages ◽  
Single Cells ◽  
Cell Movement ◽  
Time Lapse ◽  
Microscopic Investigation ◽  
Electron Microscopic Investigation ◽  
Electron Microscopic ◽  
Annual Fish ◽  
Fish Embryos

During epiboly and early dispersed stages, the deep blastomeres of annual fish embryos emigrate from the blastodisc to wander as easily visible single cells in the narrow space between the yolk synctial layer (YSL) and the enveloping cell layer (ECL). Subsequently these deep blastomeres reaggregate and eventually form the definitive embryo. We are studying these morphogen-etic cell movements and the controls that may exist upon cell movement in vivo. Time-lapse cinemicrophotography showed three types of cell movement by the deep blastomeres during epiboly through reaggregation stages [1], The present electron microscopic investigation was undertaken to reveal further details of the intercellular contacts during these developmental stages.

Biological Behaviors of Keratinocytes Cultured on Chitosan-Gelatin Membrane

2005 ◽  
Vol 288-289 ◽  
pp. 401-404 ◽  
Author(s):  
G.H. Yang ◽  
J. Yang ◽  
Jian Mei Wang ◽  
Lei Cui ◽  
Wei Liu ◽  
...  
Keyword(s):  
Human Keratinocytes ◽  
Migration Distance ◽  
Keratinocyte Proliferation ◽  
Cultured Keratinocytes ◽  
Chitosan Membrane ◽  
Forming Efficiency ◽  
And Migration ◽  
Dressing Materials

Chitin/Chitosan membrane has been used as wound dressing materials to facilitate clinical wound healing for many years. However, there are fewer articles studying the cell-biomaterial interaction in vitro or in vivo. In this study, the biological characteristics of human keratinocytes cultured on chitosan membrane that was mixed with gelatin in different ratio were investigated in vitro. Chitosan-gelatin membrane (CGM) in different ratio were prepared with N, N-(3 dimethylaminopropyl)-N'-ethyl carbodiimide (EDC). CGMs were divided into four groups: pure chitosan membrane, 7:3 (chitosan: gelatin), 5:5 and 3:7 groups. Human keratinocytes were isolated from foreskin by Dispase/Trypsin-EDTA digestion. Keratinocytes of passage3 were then seeded on the surface of CGM. Cloning forming efficiency (CFE) and migration distance of cultured keratinocytes on CGM were measured. The CFE of keratinocytes cultured on the surface of pure chitosan membrane was 9.8±2.08%; cultured on 7:3 CGM was 14.33±1.53%, 5:5 CGM was 19.17±1.26%, 3:7 CGM was 18.33±2.08%. The migration distance of cultured keratinocytes on pure chitosan membrane was 61.47±2.70µm, 7:3 CGM was 66.22±9.39µm, 5:5 CGM was 120.31±15.82µm, 3:7 CGM was 225.38±10.48µm. This sutdy demonstrated that increasing contents of gelatin in CGM could promote keratinocyte proliferation and migration. The results also suggested the membrane prepared from chitosan and gelatin can be utilized as a good keratinocyte delivery system.

scholarly journals Overexpression of the Oral Mucosa-Specific microRNA-31 Promotes Skin Wound Closure

2019 ◽  
Vol 20 (15) ◽  
pp. 3679 ◽  
Author(s):  
Lin Chen ◽  
Alyne Simões ◽  
Zujian Chen ◽  
Yan Zhao ◽  
Xinming Wu ◽  
...  
Keyword(s):  
Wound Healing ◽  
Oral Mucosa ◽  
Wound Closure ◽  
Expression Patterns ◽  
Skin Wound ◽  
Skin Wound Healing ◽  
Specific Expression ◽  
Keratinocyte Proliferation

Wounds within the oral mucosa are known to heal more rapidly than skin wounds. Recent studies suggest that differences in the microRNAome profiles may underlie the exceptional healing that occurs in oral mucosa. Here, we test whether skin wound-healing can be accelerating by increasing the levels of oral mucosa-specific microRNAs. A panel of 57 differentially expressed high expresser microRNAs were identified based on our previously published miR-seq dataset of paired skin and oral mucosal wound-healing [Sci. Rep. (2019) 9:7160]. These microRNAs were further grouped into 5 clusters based on their expression patterns, and their differential expression was confirmed by TaqMan-based quantification of LCM-captured epithelial cells from the wound edges. Of these 5 clusters, Cluster IV (consisting of 8 microRNAs, including miR-31) is most intriguing due to its tissue-specific expression pattern and temporal changes during wound-healing. The in vitro functional assays show that ectopic transfection of miR-31 consistently enhanced keratinocyte proliferation and migration. In vivo, miR-31 mimic treatment led to a statistically significant acceleration of wound closure. Our results demonstrate that wound-healing can be enhanced in skin through the overexpression of microRNAs that are highly expressed in the privileged healing response of the oral mucosa.

scholarly journals Genome-Wide Transcriptomic Identification and Functional Insight of Lily WRKY Genes Responding to Botrytis Fungal Disease

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 776
Author(s):  
Shipra Kumari ◽  
Bashistha Kumar Kanth ◽  
Ju young Ahn ◽  
Jong Hwa Kim ◽  
Geung-Joo Lee
Keyword(s):  
Abiotic Stress ◽  
Biotic Stress ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Lilium Longiflorum ◽  
Biotic And Abiotic Stress ◽  
Biotic Stresses ◽  
Genome Wide

Genome-wide transcriptome analysis using RNA-Seq of Lilium longiflorum revealed valuable genes responding to biotic stresses. WRKY transcription factors are regulatory proteins playing essential roles in defense processes under environmental stresses, causing considerable losses in flower quality and production. Thirty-eight WRKY genes were identified from the transcriptomic profile from lily genotypes, exhibiting leaf blight caused by Botrytis elliptica. Lily WRKYs have a highly conserved motif, WRKYGQK, with a common variant, WRKYGKK. Phylogeny of LlWRKYs with homologous genes from other representative plant species classified them into three groups- I, II, and III consisting of seven, 22, and nine genes, respectively. Base on functional annotation, 22 LlWRKY genes were associated with biotic stress, nine with abiotic stress, and seven with others. Sixteen unique LlWRKY were studied to investigate responses to stress conditions using gene expression under biotic and abiotic stress treatments. Five genes—LlWRKY3, LlWRKY4, LlWRKY5, LlWRKY10, and LlWRKY12—were substantially upregulated, proving to be biotic stress-responsive genes in vivo and in vitro conditions. Moreover, the expression patterns of LlWRKY genes varied in response to drought, heat, cold, and different developmental stages or tissues. Overall, our study provides structural and molecular insights into LlWRKY genes for use in the genetic engineering in Lilium against Botrytis disease.

scholarly journals Interfollicular epidermal stem-like cells for the recreation of the hair follicle epithelial compartment

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Carla M. Abreu ◽  
Rogério P. Pirraco ◽  
Rui L. Reis ◽  
Mariana T. Cerqueira ◽  
Alexandra P. Marques
Keyword(s):  
Hair Follicle ◽  
Cell Sorting ◽  
Dermal Papilla ◽  
Epidermal Keratinocytes ◽  
Sebaceous Glands ◽  
Phenotypic Profile ◽  
Cell Source ◽  
Epithelial Compartment ◽  
The Ideal

Abstract Background Hair follicle (HF) development and growth are dependent on epithelial-mesenchymal interactions (EMIs). Dermal papilla (DP) cells are recognized as the key inductive mesenchymal player, but the ideal source of receptive keratinocytes for human HF regeneration is yet to be defined. We herein investigated whether human interfollicular epidermal keratinocytes with stem-like features (EpSlKCs), characterized by a α6bri/CD71dim expression, can replace human hair follicular keratinocytes (HHFKCs) for the recreation of the HF epithelium and respective EMIs. Methods The α6bri/CD71dim cellular fraction was selected from the whole interfollicular keratinocyte population through fluorescence-activated cell sorting and directly compared with follicular keratinocytes in terms of their proliferative capacity and phenotype. The crosstalk with DP cells was studied in an indirect co-culture system, and EpSlKC hair forming capacity tested in a hair reconstitution assay when combined with DP cells. Results EpSlKCs exhibited a phenotypic profile similar to follicular keratinocytes and were capable of increasing DP cell proliferation and, for short co-culture times, the number of alkaline phosphatase-active cells, suggesting an improvement of their inductivity. Moreover, the recreation of immature HFs and sebaceous glands was observed after EpSlKC and DP cell co-grafting in nude mice. Conclusions Our results suggest that EpSlKCs are akin to follicular keratinocytes and can crosstalk with DP cells, contributing to HF morphogenesis in vivo, thus representing an attractive epithelial cell source for hair regeneration strategies.

scholarly journals Single cell transcriptome atlas of mouse mammary epithelial cells across development

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Bhupinder Pal ◽  
Yunshun Chen ◽  
Michael J. G. Milevskiy ◽  
François Vaillant ◽  
Lexie Prokopuk ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Developmental Stages ◽  
Mammary Epithelial Cells ◽  
Expression Profiles ◽  
Single Cells ◽  
Chromatin Accessibility ◽  
Mammary Epithelial ◽  
Cell Transcriptome ◽  
Single Cell Transcriptome

Abstract Background Heterogeneity within the mouse mammary epithelium and potential lineage relationships have been recently explored by single-cell RNA profiling. To further understand how cellular diversity changes during mammary ontogeny, we profiled single cells from nine different developmental stages spanning late embryogenesis, early postnatal, prepuberty, adult, mid-pregnancy, late-pregnancy, and post-involution, as well as the transcriptomes of micro-dissected terminal end buds (TEBs) and subtending ducts during puberty. Methods The single cell transcriptomes of 132,599 mammary epithelial cells from 9 different developmental stages were determined on the 10x Genomics Chromium platform, and integrative analyses were performed to compare specific time points. Results The mammary rudiment at E18.5 closely aligned with the basal lineage, while prepubertal epithelial cells exhibited lineage segregation but to a less differentiated state than their adult counterparts. Comparison of micro-dissected TEBs versus ducts showed that luminal cells within TEBs harbored intermediate expression profiles. Ductal basal cells exhibited increased chromatin accessibility of luminal genes compared to their TEB counterparts suggesting that lineage-specific chromatin is established within the subtending ducts during puberty. An integrative analysis of five stages spanning the pregnancy cycle revealed distinct stage-specific profiles and the presence of cycling basal, mixed-lineage, and 'late' alveolar intermediates in pregnancy. Moreover, a number of intermediates were uncovered along the basal-luminal progenitor cell axis, suggesting a continuum of alveolar-restricted progenitor states. Conclusions This extended single cell transcriptome atlas of mouse mammary epithelial cells provides the most complete coverage for mammary epithelial cells during morphogenesis to date. Together with chromatin accessibility analysis of TEB structures, it represents a valuable framework for understanding developmental decisions within the mouse mammary gland.

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