A novel method of neural differentiation of PC12 cells by using Opti-MEM as a basic induction medium

Hu, Ren; Cao, Qiaoyu; Sun, Zhen; Chen, Jinying; Zheng, Qi; Xiao, Fei

doi:10.3892/ijmm.2017.3195

Cited by 44 publications

(45 citation statements)

References 41 publications

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“…The mutations P206S, R303W, and R441H have already been reported to reduce TPH2 activity [11]. However, an in vitro study using PC12 cells, a cell line derived from a pheochromocytoma of the rat adrenal medulla [37], showed that the mutation R441H does not impair TPH2 oligomerization [36]. Thus, to further investigate the impact of P206S, R303W, and R441H amino acid substitutions on dimer coordination, we performed MM-PBSA analysis of TPH2 WT and its variants, as well as analyzed the number of hydrogen bonds and hydrophobic contacts formed between the TPH2 monomers throughout the simulations.…”

Section: Plos Onementioning

confidence: 99%

In silico analysis of the tryptophan hydroxylase 2 (TPH2) protein variants related to psychiatric disorders

et al. 2020

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The tryptophan hydroxylase 2 (TPH2) enzyme catalyzes the first step of serotonin biosynthesis. Serotonin is known for its role in several homeostatic systems related to sleep, mood, and food intake. As the reaction catalyzed by TPH2 is the rate-limiting step of serotonin biosynthesis, mutations in TPH2 have been associated with several psychiatric disorders (PD). This work undertakes an in silico analysis of the effects of genetic mutations in the human TPH2 protein. Ten algorithms were used to predict the functional and stability effects of the TPH2 mutations. ConSurf was used to estimate the evolutionary conservation of TPH2 amino acids. GROMACS was used to perform molecular dynamics (MD) simulations of TPH2 WT and P260S, R303W, and R441H, which had already been associated with the development of PD. Forty-six TPH2 variants were compiled from the literature. Among the analyzed variants, those occurring at the catalytic domain were shown to be more damaging to protein structure and function. The ConSurf analysis indicated that the mutations affecting the catalytic domain were also more conserved throughout evolution. The variants S364K and S383F were predicted to be deleterious by all the functional algorithms used and occurred at conserved positions, suggesting that they might be deleterious. The MD analyses indicate that the mutations P206S, R303W, and R441H affect TPH2 flexibility and essential mobility at the catalytic and oligomerization domains. The variants P206S, R303W, and R441H also exhibited alterations in dimer binding affinity and stability throughout the simulations. Thus, these mutations may impair TPH2 functional interactions and, consequently, its function, leading to the development of PD. Furthermore, we developed a database, SNPMOL (http://www.snpmol.org/), containing the results presented in this paper. Understanding the effects of TPH2 mutations on protein structure and function may lead to improvements in existing treatments for PD and facilitate the design of further experiments.

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Section: Plos Onementioning

confidence: 99%

In silico analysis of the tryptophan hydroxylase 2 (TPH2) protein variants related to psychiatric disorders

et al. 2020

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“…The generation of action potentials is an important physiological property of mature neurons [ 29 , 30 ]. To assess whether action potentials are generated in NSC-34 cells, we performed current-clamp recordings on non-neurite-bearing cells treated with a vehicle for 2 days and neurite-bearing cells treated with PGE 2 (30 μM) for 2 days or RA (10 μM) for 7 days.…”

Section: Resultsmentioning

confidence: 99%

Highly Efficient Conversion of Motor Neuron-Like NSC-34 Cells into Functional Motor Neurons by Prostaglandin E2

Nango

Kosuge

Sato

et al. 2020

Cells

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Motor neuron diseases are a group of progressive neurological disorders that degenerate motor neurons. The neuroblastoma × spinal cord hybrid cell line NSC-34 is widely used as an experimental model in studies of motor neuron diseases. However, the differentiation efficiency of NSC-34 cells to neurons is not always sufficient. We have found that prostaglandin E2 (PGE2) induces morphological differentiation in NSC-34 cells. The present study investigated the functional properties of PGE2-differentiated NSC-34 cells. Retinoic acid (RA), a widely-used agent inducing cell differentiation, facilitated neuritogenesis, which peaked on day 7, whereas PGE2-induced neuritogenesis took only 2 days to reach the same level. Whole-cell patch-clamp recordings showed that the current threshold of PGE2-treated cell action potentials was lower than that of RA-treated cells. PGE2 and RA increased the protein expression levels of neuronal differentiation markers, microtubule-associated protein 2c and synaptophysin, and to the same extent, motor neuron-specific markers HB9 and Islet-1. On the other hand, protein levels of choline acetyltransferase and basal release of acetylcholine in PGE2-treated cells were higher than in RA-treated cells. These results suggest that PGE2 is a rapid and efficient differentiation-inducing factor for the preparation of functionally mature motor neurons from NSC-34 cells.

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“…It was established as a neuron study model early in 1980. [ 46 ] Most studies have investigated the morphology of PC12 cells on electrospun nanofibers rather than gels. The adhesion and proliferation of these cells on PCL/chitosan/polypyrrole electrospun nanofibers was higher than that of pure PCL nanofibers.…”

Section: Discussionmentioning

confidence: 99%

Bilayer Cylindrical Conduit Consisting of Electrospun Polycaprolactone Nanofibers and DSC Cross‐Linked Sodium Alginate Hydrogel to Bridge Peripheral Nerve Gaps

Askarzadeh

Nazarpak

Mansoori

et al. 2020

Macromolecular Bioscience

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Herein, a bilayer cylindrical conduit (P‐CA) is presented consisting of electrospun polycaprolactone (PCL) nanofibers and sodium alginate hydrogel covalently cross‐linked with N,N′‐disuccinimidyl carbonate (DSC). The bilayer P‐CA conduit is developed by combining the electrospinning and outer–inner layer methods. Using DSC, as a covalent crosslinker, increases the degradation time of the sodium alginate hydrogel up to 2 months. The swelling ratio of the hydrogel is also 503% during the first 8 h. The DSC cross‐linked sodium alginate in the inner layer of the conduit promotes the adhesion and proliferation of nerve cells, while the electrospun PCL nanofibers in the outer layer provide maximum tensile strength of the conduit during surgery. P‐CA conduit promotes the migration of Schwann cells along the axon in a rat model based on functional and histological evidences. In conclusion, P‐CA conduit will be a promising construct for repairing sciatic nerves in a rat model.

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A novel method of neural differentiation of PC12 cells by using Opti-MEM as a basic induction medium

Cited by 44 publications

References 41 publications

In silico analysis of the tryptophan hydroxylase 2 (TPH2) protein variants related to psychiatric disorders

In silico analysis of the tryptophan hydroxylase 2 (TPH2) protein variants related to psychiatric disorders

Highly Efficient Conversion of Motor Neuron-Like NSC-34 Cells into Functional Motor Neurons by Prostaglandin E2

Bilayer Cylindrical Conduit Consisting of Electrospun Polycaprolactone Nanofibers and DSC Cross‐Linked Sodium Alginate Hydrogel to Bridge Peripheral Nerve Gaps

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