Articles by Agnes S. Pascual in JoVE
Zebrafish Keratocyte Explants to Study Collective Cell Migration and Reepithelialization in Cutaneous Wound Healing Jose L. Rapanan1, Agnes S. Pascual1, Chandana K. Uppalapati2, Kimbal E. Cooper1, Kathryn J. Leyva2, Elizabeth E. Hull1 1Biomedical Sciences Program, Midwestern University, 2Department of Microbiology & Immunology, Midwestern University Zebrafish keratocytes migrate in cell sheets from explants and provide an in vitro model for the study of the mechanisms of collective cell migration in the context of epithelial wound healing. These protocols detail an effective way to establish primary explant cultures for use in collective cell migration assays.
Establishment of a Clinic-based Biorepository Sarah E. Belden1, Chandana K. Uppalapati2, Agnes S. Pascual3, McKale R. Montgomery3, Kathryn J. Leyva2, Elizabeth E. Hull3, Richard L. Averitte1 1Affiliated Dermatology & Affiliated Laboratories, Midwestern University Osteopathic Postdoctoral Training Institute, Midwestern University, 2Department of Microbiology & Immunology, Arizona College of Osteopathic Medicine, Midwestern University, 3Biomedical Sciences Program, College of Health Sciences, Midwestern University Cutaneous tumors are often discarded following Mohs micrographic surgery. A protocol is described here that enables clinical support staff to effectively process and store cutaneous tumor (e.g., squamous cell carcinoma, basal cell carcinoma, and melanoma) samples for downstream laboratory applications without interfering with clinical operations.
Other articles by Agnes S. Pascual on PubMed
Zebrafish Keratocyte Explant Cultures As a Wound Healing Model System: Differential Gene Expression & Morphological Changes Support Epithelial-mesenchymal Transition Experimental Cell Research. Jul, 2013 | Pubmed ID: 23588205 The control of collective cell migration of zebrafish keratocyte sheets in explant culture is of interest for cell migration and epithelial wound healing and depends on the gene expression profile. In a zebrafish genome array, ∼17.5% of the probe sets were differentially expressed greater than two-fold (p≤0.003) between 1 and 7 days of explant culture. Among the differentially expressed genes were a variety of wound healing-related genes and many of the biomarkers for epithelial-mesenchymal transition (EMT), including a switch from keratin and E-cadherin to vimentin and N-cadherin expression and several EMT-related transcription factors were found to be differentially expressed. Supporting evidence for EMT is seen in both morphological change and rearrangement of the actin cytoskeleton and in expression of cadherins during explant culture with a visible disassembly of the cell sheet. TGFβ1 and TNFα expression were analyzed by qPCR at various time points and peak differential expression of both cytokines occurred at 3 days, indicating that the EMT process is ongoing under conditions routinely used in the study of fish keratocyte motility. These data establish that an EMT process is occurring during zebrafish keratocyte explant culture and support the use of this system as a wound healing model.
Epigenetically Maintained SW13+ and SW13- Subtypes Have Different Oncogenic Potential and Convert with HDAC1 Inhibition BMC Cancer. May, 2016 | Pubmed ID: 27188282 The BRM and BRG1 tumor suppressor genes are mutually exclusive ATPase subunits of the SWI/SNF chromatin remodeling complex. The human adrenal carcinoma SW13 cell line can switch between a subtype which expresses these subunits, SW13+, and one that expresses neither subunit, SW13-. Loss of BRM expression occurs post-transcriptionally and can be restored via histone deacetylase (HDAC) inhibition. However, most previously used HDAC inhibitors are toxic and broad-spectrum, providing little insight into the mechanism of the switch between subtypes. In this work, we explore the mechanisms of HDAC inhibition in promoting subtype switching and further characterize the oncogenic potential of the two epigenetically distinct SW13 subtypes.