Chromatin-binding HMGN proteins and the neuronal differentiation of enbryonal carcinoma cells in vitro

Mohan, Gokula (2012) Chromatin-binding HMGN proteins and the neuronal differentiation of enbryonal carcinoma cells in vitro. PhD thesis, University of Glasgow.

Abstract

Embryonic stem (ES) cells are able to differentiate in vitro into endodermal, mesodermal and ectodermal cell types. ES cells and their close counterparts, embryonic carcinoma (EC) cells, are a useful model system for studying the mechanisms governing neuronal differentiation. Since High Mobility Group-nucleosome binding (HMGN) genes are regulated in a developmental-stage specific manner during mouse embryogenesis and cellular differentiation, their roles in undifferentiated and neural differentiating P19 EC cells were examined. Work presented in this thesis firstly optimises the Retinoic Acid (RA) -induced neural differentiation protocol of P19 EC cells based on key neuronal and glia markers. Two crucial steps of RA concentration and cell plating density were shown to increase the efficiency of neuronal differentiation. Analysis of HMGN proteins showed they were ubiquitously expressed in undifferentiated and neural differentiating P19 cells. HMGN2 and HMGN3 were up-regulated while HMGN1 remained unchanged upon neural commitment. Unusually, HMGN3 protein was localised in the cytoplasm of P19 cells. To study the possible role of HMGN proteins, HMGN1 and HMGN2 were knocked down using siRNAs. HMGN1 and HMGN2 knockdown in undifferentiated P19 EC cells dramatically down-regulated the key pluripotency regulator genes Oct4, Nanog and Sox2. Furthermore, HMGN1 and HMGN2 knockdown in neural differentiating cells affected seven neuron-specific genes. These data suggest that HMGN proteins may play roles in regulating genes that are involved in maintaining pluripotency and regulating neural differentiation in P19 cells.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Keywords: HMGN,embryonic stem cells,neuronal differentiation,pluripotency,chromatin
• Subjects: R Medicine > R Medicine (General); Q Science > Q Science (General)
• Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Supervisor's Name: West, Dr. Katherine
• Date of Award: 2012
• Depositing User: Mr Gokula Mohan
• Unique ID: glathesis:2012-3316
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 18 Apr 2012
• Last Modified: 10 Dec 2012 14:06
• URI: https://theses.gla.ac.uk/id/eprint/3316