PUBLICATION

Developmental regulation of TAC1 in peptidergic-induced human mesenchymal stem cells: Implication for spinal cord injury in zebrafish

Authors
Patel, N., Klassert, T., Greco, S.J., Patel, S.A., Munoz, J.L., Reddy, B.Y., Bryan, M., Campbell, N., Kokorina, N., Sabaawy, H.E., and Rameshwar, P.
ID
ZDB-PUB-110628-27
Date
2012
Source
Stem cells and development   21(2): 308-320 (Journal)
Registered Authors
Campbell, Neil, Sabaawy, Hatem
Keywords
none
MeSH Terms
  • Activating Transcription Factors/genetics
  • Activating Transcription Factors/metabolism
  • Animals
  • Binding Sites
  • Cells, Cultured
  • Co-Repressor Proteins
  • Cyclic AMP Response Element-Binding Protein/genetics
  • Cyclic AMP Response Element-Binding Protein/metabolism
  • Fibroblast Growth Factor 2/pharmacology
  • Gene Expression Regulation*/drug effects
  • Genes, Reporter
  • Humans
  • Models, Biological
  • Nerve Tissue Proteins/genetics
  • Nerve Tissue Proteins/metabolism*
  • Protein Binding
  • Proto-Oncogene Proteins c-jun/genetics
  • Proto-Oncogene Proteins c-jun/metabolism
  • Repressor Proteins/genetics
  • Repressor Proteins/metabolism*
  • Response Elements/genetics
  • Signal Transduction/drug effects
  • Signal Transduction/genetics*
  • Spinal Cord Injuries/metabolism
  • Spinal Cord Injuries/pathology
  • Spinal Cord Injuries/therapy*
  • Tachykinins/genetics
  • Tachykinins/metabolism*
  • Transcription Factor AP-1/genetics
  • Transcription Factor AP-1/metabolism
  • Transplantation, Heterologous
  • Tretinoin/pharmacology
  • Zebrafish
PubMed
21671725 Full text @ Stem Cells Dev.
Abstract
Human mesenchymal stem cells (MSCs) are easy to expand, relatively safe and can be transplanted in allogeneic recipients as `off-the-shelf' cells. MSCs can be induced to form functional peptidergic neurons and express the neurotransmitter gene, TAC1. The expression of TAC1 requires that the repressor gene, REST, is decreased. This study investigated the molecular pathway in TAC1 induction as MSCs differentiated into neurons and then applied the findings in a model of spinal cord injury (SCI) in zebrafish. We studied the developmental roles of the two cyclic AMP response element (CRE) sites: CRE1 and CRE2. AP-1 binding site overlaps with CRE2 (CRE2/AP-1). Reporter gene studies with the 5' regulatory region of TAC1 containing wild-type or mutant CRE sites and, parallel studies with ectopically expressed inhibitor of cAMP proteins (ICER) indicated that CRE1 and CRE2/AP-1 are activated at days 6 and 12, respectively. Studies with PKA and JNK inhibitors in the reporter gene studies, chromatin immunoprecipation assay and ectopic expression of REST indicated the following pathways: Decrease of REST activated upstream JNK. In turn, JNK activated ATF-2 and AP-1 for interaction with CRE1 and CRE2/AP-1, respectively. In order to apply the finding to SCI, we transplanted 6-day induced MSCs in transgenic HB9-GFP zebrafish larvae with SCI, in the presence or absence of JNK inhibitors. Imaging and functional studies showed significant improvement in the fish. The repair mechanism involved the activation of JNK. The findings have long-term implications for SCI repair with MSCs.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping