title: Prion protein expression regulates embryonic stem cell pluripotency and differentiation.
abstract: Cellular prion protein (PRNP) is a glycoprotein involved in the pathogenesis of transmissible spongiform encephalopathies (TSEs). Although the physiological function of PRNP is largely unknown, its key role in prion infection has been extensively documented. This study examines the functionality of PRNP during the course of embryoid body (EB) differentiation in mouse Prnp-null (KO) and WT embryonic stem cell (ESC) lines. The first feature observed was a new population of EBs that only appeared in the KO line after 5 days of differentiation. These EBs were characterized by their expression of several primordial germ cell (PGC) markers until Day 13. In a comparative mRNA expression analysis of genes playing an important developmental role during ESC differentiation to EBs, Prnp was found to participate in the transcription of a key pluripotency marker such as Nanog. A clear switching off of this gene on Day 5 was observed in the KO line as opposed to the WT line, in which maximum Prnp and Nanog mRNA levels appeared at this time. Using a specific antibody against PRNP to block PRNP pathways, reduced Nanog expression was confirmed in the WT line. In addition, antibody-mediated inhibition of ITGB5 (integrin αvβ5) in the KO line rescued the low expression of Nanog on Day 5, suggesting the regulation of Nanog transcription by Prnp via this Itgb5. mRNA expression analysis of the PRNP-related proteins PRND (Doppel) and SPRN (Shadoo), whose PRNP function is known to be redundant, revealed their incapacity to compensate for the absence of PRNP during early ESC differentiation. Our findings provide strong evidence for a relationship between Prnp and several key pluripotency genes and attribute Prnp a crucial role in regulating self-renewal/differentiation status of ESC, confirming the participation of PRNP during early embryogenesis.
authors.affiliation: Departamento de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain. miranda.alberto@inia.es
authors.forename: Alberto
authors.initials: A
authors.lastname: Miranda
authors.forename: Eva
authors.initials: E
authors.lastname: Pericuesta
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Ramírez
authors.forename: Alfonso
authors.initials: A
authors.lastname: Gutierrez-Adan
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D015536:Down-Regulation / Q000235:genetics; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D047108:Embryonic Development / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D018398:Homeodomain Proteins / Q000235:genetics; D039641:Integrin beta Chains / Q000235:genetics; D008297:Male; D051379:Mice; D000071317:Nanog Homeobox Protein; D010053:Ovary / Q000254:growth & development / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D011328:Prions / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000235:genetics; D013737:Testis / Q000254:growth & development / Q000378:metabolism; D013997:Time Factors; D015854:Up-Regulation / Q000235:genetics
chemical_list: D015415:Biomarkers; D018398:Homeodomain Proteins; D039641:Integrin beta Chains; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D011328:Prions; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor; C084344:integrin beta5