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cAMP initiates early phase neuron-like morphology changes and late phase neural differentiation in mesenchymal stem cells

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Abstract

The intracellular second messenger cAMP is frequently used in induction media to induce mesenchymal stem cells (MSCs) into neural lineage cells. To date, an understanding of the role cAMP exerts on MSCs and whether cAMP can induce MSCs into functional neurons is still lacking. We found cAMP initiated neuron-like morphology changes early and neural differentiation much later. The early phase changes in morphology were due to cell shrinkage, which subsequently rendered some cells apoptotic. While the morphology changes occurred prior to the expression of neural markers, it is not required for neural marker expression and the two processes are differentially regulated downstream of cAMP-activated protein kinase A. cAMP enabled MSCs to gain neural marker expressions with neuronal function, such as, calcium rise in response to neuronal activators, dopamine, glutamate, and potassium chloride. However, only some of the cells induced by cAMP responded to the three neuronal activators and further lack the neuronal morphology, suggesting that although cAMP is able to direct MSCs towards neural differentiation, they do not achieve terminal differentiation.

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Abbreviations

cAMP:

Cyclic adenosine monophosphate

MSCs:

Mesenchymal stem cells

PKA:

Protein kinase A

PKAc:

PKA catalytic subunit

pPKAc:

Threonine 197 phosphorylated PKAc

SCI:

Spinal cord injury

IBMX:

Isobutylmethylxanthine

FI:

Forskolin and IBMX

ActD:

Actinomycin D

CHX:

Cycloheximide

Ptx:

Paclitaxel

PI:

Propidium iodide

NSE:

Neuron-specific enolase

Tuj1:

β-III Tubulin

GFAP:

Glial fibrillary acidic protein

CREB:

cAMP response element binding protein

pCREB:

Serine 133 phosphorylated CREB

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Acknowledgements

We thank Dr. Carlose Molina at New Jersey Medical School for kindly providing the ICER antibody and Dr. David Ginty at Johns Hopkins University for kindly providing the M1-CREB plasmid. This study was supported in part by the National Science Foundation (CBET 0941055), the National Institute of Health (R01GM079688 and R21RR024439), the MUCI, and the MSU Foundation and the Center for Systems Biology.

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Authors and Affiliations

  1. Department of Chemical Engineering and Materials Science, Michigan State University, 2527 EB, East Lansing, MI, 48824, USA

    Linxia Zhang, Linsey C. Seitz, Amy M. Abramczyk & Christina Chan

  2. Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, USA

    Christina Chan

  3. Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, USA

    Li Liu

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  1. Linxia Zhang
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Correspondence to Christina Chan.

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Zhang, L., Seitz, L.C., Abramczyk, A.M. et al. cAMP initiates early phase neuron-like morphology changes and late phase neural differentiation in mesenchymal stem cells. Cell. Mol. Life Sci. 68, 863–876 (2011). https://doi.org/10.1007/s00018-010-0497-1

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