Impact of Reck expression and promoter activity in neuronal in vitro differentiation

Author(s)

Trombetta-Lima, Marina ^[1] ; Assis-Ribas, Thais ^[1] ; Cintra, Ricardo C. ^[2] ; Campeiro, Joana D. ^[3] ; Guerreiro, Juliano R. ^[4] ; Winnischofer, Sheila M. B. ^{[5, 6]} ; Nascimento, Isis C. C. ^[2] ; Ulrich, Henning ^[2] ; Hayashi, Mirian A. F. ^[3] ; Sogayar, Mari C. ^{[2, 1]}

Affiliation

^[1] Univ Sao Paulo, Fac Med, Nucleo Terapia Celular & Mol NUCEL, Rua Pangare 100, BR-05360130 Sao Paulo, SP - Brazil

^[2] Univ Sao Paulo, Inst Quim, Dept Bioquim, BR-05508000 Sao Paulo, SP - Brazil

^[3] Univ Fed Sao Paulo UNIFESP, Escola Paulista Med EPM, Dept Farmacol, Rua 3 Maio 100, Ed INFAR, 3 Andar, BR-04044020 Sao Paulo, SP - Brazil

^[4] Univ Paulista UNIP, Fac Farm, BR-05347020 Sao Paulo, SP - Brazil

^[5] Univ Fed Parana UFPR, Dept Bioquim & Biol Mol, BR-81531990 Curitiba, PR - Brazil

^[6] Univ Fed Parana UFPR, Dept Biol Celular, BR-81531990 Curitiba, PR - Brazil

• Document type: Journal article
• Source: MOLECULAR BIOLOGY REPORTS; v. 48, n. 2 FEB 2021.
• Web of Science Citations: 0

Abstract

Reck (REversion-inducing Cysteine-rich protein with Kazal motifs) tumor suppressor gene encodes a multifunctional glycoprotein which inhibits the activity of several matrix metalloproteinases (MMPs), and has the ability to modulate the Notch and canonical Wnt pathways. Reck-deficient neuro-progenitor cells undergo precocious differentiation; however, modulation of Reck expression during progression of the neuronal differentiation process is yet to be characterized. In the present study, we demonstrate that Reck expression levels are increased during in vitro neuronal differentiation of PC12 pheochromocytoma cells and P19 murine teratocarcinoma cells and characterize mouse Reck promoter activity during this process. Increased Reck promoter activity was found upon induction of differentiation in PC12 cells, in accordance with its increased mRNA expression levels in mouse in vitro models. Interestingly, Reck overexpression, prior to the beginning of the differentiation protocol, led to diminished efficiency of the neuronal differentiation process. Taken together, our findings suggest that increased Reck expression at early stages of differentiation diminishes the number of neuron-like cells, which are positive for the beta-3 tubulin marker. Our data highlight the importance of Reck expression evaluation to optimize in vitro neuronal differentiation protocols. (AU)

• FAPESP's process: 18/20014-0 - The gut-brain connection: role of the gut microbioma in the outcome and development of the sporadic Parkinson's disease
• Grantee: Matheus de Castro Fonseca
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/05311-2 - Regenerative medicine aiming at therapy for chronic degenerative diseases (cancer and diabetes)
• Grantee: Mari Cleide Sogayar
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 18/07366-4 - Purine and kinin receptors as targets of study and therapeutic interventions in neurological diseases
• Grantee: Alexander Henning Ulrich
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/50891-1 - INCT 2014: Translational Medicine
• Grantee: Jaime Eduardo Cecilio Hallak
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 19/13112-8 - Study of molecular and cellular mechanisms involved in mental disorders: clinical and animal models analysis
• Grantee: Mirian Akemi Furuie Hayashi
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 15/26328-8 - Comparative analysis of the Reck tumor suppressor gene role in glioblastoma stem cell sub-population and in normal neural stem cells.
• Grantee: Marina Trombetta Lima
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 16/18277-7 - ANALYSIS OF THE RECK TUMOR SUPPRESSOR GENE'S ROLE IN ASYMMETRIC CELL DIVISION
• Grantee: Marina Trombetta Lima
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 12/50880-4 - Stem cells: from basic studies of kinin and purinergic receptor roles towards therapeutical applications
• Grantee: Alexander Henning Ulrich
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/02413-1 - Validation of crotamine as a biomarker and evaluation of its potential use in the therapy of human diseases
• Grantee: Mirian Akemi Furuie Hayashi
• Support Opportunities: Regular Research Grants