Abstract
Central nervous system (CNS) diseases can cause a series of neuronal lesions, which may be improved by the anti-apoptotic neuroprotection of transforming growth factor-beta 1 (TGF-β1). In neurons, L-type Ca2+ channels (LTCC) are mainly composed of Cav1.2 subunits. Given the implication of TGF-β1 in numerous CNS diseases, we examined the neuroprotective effects of TGF-β1 on the Cav1.2 channel in the CNS. To simulate acute mechanical traumatic brain injury (TBI), we used a needle to create parallel scratches across plates, which were cultured for 9 h. Meanwhile, Fluo4-AM-loaded laser scanning confocal microscopy with a dual wavelength of 488 nm/530 nm was employed to determine intracellular calcium concentrations ([Ca2+]i). We found that MAPK inhibitors impede TGF-β1-induced cell viability and that TGF-β1 recovered from the trauma-induced cell viability in neurons. Cav1.2 production was significantly decreased in the TGF-β1-treated (10 ng/mL) neurons. At this TGF-β1 concentration, Cav1.2 was significantly down-regulated in a time-dependent manner after 12 h. Moreover, TGF-β1 partially recovered the protein levels of Cav1.2 that were reduced by TBI. TGF-β1 significantly inhibited the fluorescence intensity of [Ca2+]i increased by KCl and delayed the time of the peak [Ca2+]i. The observed effects of TGF-β1 on Cav1.2 were regulated by MAPK inhibitors. The observed effects of TGF-β1 on P-JNK were also impeded by pre-incubation with the LTCC inhibitor (10 μM) nimodipine in trauma-injured neurons. Altogether, TGF-β1 regulated LTCCs through a mechanism dependent on MEK, JNK1/2 and p38 MAPK signal pathways in cortical neurons. Thus, we suggest the involvement of this mechanism in cell viability.
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Abbreviations
- TGF-β1:
-
transforming growth factor-beta1
- TGF-βRI:
-
transforming growth factor-beta1 type I receptor
- MEK:
-
mitogen-activated protein kinase
- P38 MAPK:
-
P38 mitogen-activated protein kinases
- JNK1/2:
-
Jun NH2-terminal kinase 1/2
- CNS:
-
central nervous system
- LTCC:
-
L-type calcium channel
- TBI:
-
traumatic brain injury
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Acknowledgments
This project was supported by the National Natural Science Foundation of China (31170852, 31300761, 81001322, 81001340, 81172795, 81374040 and 81471622), Guangdong Natural Science Foundation (S2012040007470), the Medical Scientific Research Foundation of Guangdong Province, China (A2015211), the Foundation for Distinguished Young Teachers in Higher Education of Guangdong (Yq2013077 and Yq2013079) and the Department of Education, Guangdong Government under the Top-tier University Development Scheme for Research and Control of Infectious Diseases (2015026). The funding bodies did not participate in the study design; data collection, analysis and interpretation; and manuscript writing.
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Z.N.L. designed and performed the experiments, interpreted the data and drafted the manuscript. J.T.S. helped design the experiments and assisted with the immunoblots. G.Y.P. and J.H.Y. assisted with the cell culture and performed viability experiments. W.Q.C. and K.S.L. helped design the experiments, interpret the data and write the manuscript. All authors read and approved the final version of the manuscript.
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Animal protocols were designed in accordance with the NIH Guide for the Care and Use of Laboratory Animals and approved by the Ethics Committee of Shantou University Medical College.
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Liu, Z., Sheng, J., Peng, G. et al. TGF-β1 Regulation of P-JNK and L-Type Calcium Channel Cav1.2 in Cortical Neurons. J Mol Neurosci 64, 374–384 (2018). https://doi.org/10.1007/s12031-018-1033-2
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DOI: https://doi.org/10.1007/s12031-018-1033-2