, Volume 16, Issue 3, pp 901–911 | Cite as

Increased TRPM4 Activity in Cerebral Artery Myocytes Contributes to Cerebral Blood Flow Reduction After Subarachnoid Hemorrhage in Rats

  • Yi Gong
  • Ming-yue Du
  • Hua-lin Yu
  • Zhi-yong Yang
  • Yu-jin Li
  • Lei Zhou
  • Rong Mei
  • Li Yang
  • Fei WangEmail author
Original Article


Cerebral blood flow (CBF) reduction underlies unfavorable outcomes after subarachnoid hemorrhage (SAH). Transient receptor potential melastatin-4 (TRPM4) has a pivotal role in cerebral artery myogenic tone maintenance and CBF regulation under physiological conditions. However, the role of TRPM4 in CBF reduction after SAH is unclear. In this study, we aimed at testing whether TRPM4 would contribute to CBF reduction after SAH in vivo and determining underlying mechanisms. Rat SAH model was established by stereotaxic injection of autologous nonheparinized arterial blood at the suprasellar cistern. A TRPM4 blocker, 9-phenanthrol (9-Phe), was infused through an intraventricular catheter connected to a programmed subcutaneous pump to evaluate the contribution of TRPM4 to SAH outcomes. TRPM4 expression and translocation in cerebral artery myocytes were detected by immunoblotting. Macroscopic currents in cerebral artery myocytes were determined by whole-cell patch clamp. Myogenic tone of cerebral arteries was studied by pressurized myography. Cortical and global CBFs were measured via laser Doppler flowmetry and fluorescent microspheres, respectively. After SAH, TRPM4 translocation and macroscopic current density increased significantly. Furthermore, TRPM4 accounted for a greater proportion of myogenic tone after SAH, suggesting an upregulation of TRPM4 activity in response to SAH. Cortical and global CBFs were reduced after SAH, but were restored significantly by 9-Phe, implying that TRPM4 contributed to CBF reduction after SAH. Collectively, these discoveries show that increased TRPM4 activity has a pivotal role in CBF reduction after SAH, and provide a novel target for the management of cerebral perfusion dysfunction following SAH.

Key Words

Subarachnoid hemorrhage transient receptor potential melastatin-4 myogenic tone cerebral vasospasm cerebral blood flow. 



The authors thank Dr. Siu-Lung Chan for critically review and intellectual input on this manuscript. This work was supported by the National Natural Science Foundation of China (Nos. 81760223, 81560206), Natural Science Foundation of Yunnan Province (Nos. FB2016121, 2014FB087), Yunnan Health Training Project in High Level Talents (No. H-201601), Technology and Science Innovation Team Foundation of Kunming Medical University (No. CXTD201707), Yunnan Key Laboratory of Medicine Funding (No. 2017DG005), and Internal Funding of Yunnan Provincial Health and Family Planning Commission (No. 2016NS205).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Required Author Forms

Disclosure forms provided by the authors are available with the online version of this article.

Supplementary material

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13311_2019_741_MOESM3_ESM.pdf (490 kb)
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  1. 1.
    Dasenbrock HH, Angriman F, Smith TR, et al. Readmission after aneurysmal subarachnoid hemorrhage: a nationwide readmission database analysis. Stroke 2017; 48(9): 2383–2390.Google Scholar
  2. 2.
    Kang DW. Intracerebral hemorrhage: large disease burden but less therapeutic progress. J Stroke 2017; 19(1): 1–2.Google Scholar
  3. 3.
    Rostami E, Engquist H, Johnson U, et al. Monitoring of cerebral blood flow and metabolism bedside in patients with subarachnoid hemorrhage - A Xenon-CT and microdialysis study. Front Neurol 2014; 5: 89.Google Scholar
  4. 4.
    Tso MK, Macdonald RL. Subarachnoid hemorrhage: a review of experimental studies on the microcirculation and the neurovascular unit. Transl Stroke Res 2014; 5(2): 174–189.Google Scholar
  5. 5.
    Bauer AM, Rasmussen PA. Treatment of intracranial vasospasm following subarachnoid hemorrhage. Front Neurol 2014; 5: 72.Google Scholar
  6. 6.
    Wang F, Yin YH, Jia F, Jiang JY. Antagonism of R-type calcium channels significantly improves cerebral blood flow after subarachnoid hemorrhage in rats. J Neurotraum 2010; 27(9): 1723–1732.Google Scholar
  7. 7.
    Szarka N, Amrein K, Horvath P, et al. Hypertension-induced enhanced myogenic constriction of cerebral arteries is preserved after traumatic brain injury. J Neurotraumal 2017; 34(14): 2315–2319.Google Scholar
  8. 8.
    Simms BA, Zamponi GW. Neuronal voltage-gated calcium channels: structure, function, and dysfunction. Neuron 2014; 82(1): 24–45.Google Scholar
  9. 9.
    Guo J, She J, Zeng W, Chen Q, Bai XC, Jiang Y. Structures of the calcium-activated, non-selective cation channel TRPM4. Nature 2017; 552(7684): 205–209.Google Scholar
  10. 10.
    Earley S, Waldron BJ, Brayden JE. Critical role for transient receptor potential channel TRPM4 in myogenic constriction of cerebral arteries. Circ Res 2004; 95(9): 922–929.Google Scholar
  11. 11.
    Reading SA, Brayden JE. Central role of TRPM4 channels in cerebral blood flow regulation. Stroke 2007; 38(8): 2322–2328.Google Scholar
  12. 12.
    Ding XQ, Ban T, Liu ZY, et al. Transient receptor potential melastatin 4 (TRPM4) contributes to high salt diet-mediated early-stage endothelial injury. Cell Physiol Biochem 2017; 41(2): 835–848.Google Scholar
  13. 13.
    Wang F, Koide M, Wellman GC. Nifedipine inhibition of high-voltage activated calcium channel currents in cerebral artery myocytes is influenced by extracellular divalent cations. Front Physiol 2017; 8: 210–220.Google Scholar
  14. 14.
    Crnich R, Amberg GC, Leo MD, et al. Vasoconstriction resulting from dynamic membrane trafficking of TRPM4 in vascular smooth muscle cells. Am J Physiol Cell Physiol 2010; 299(3): C682–694.Google Scholar
  15. 15.
    Syed AU, Koide M, Brayden JE, Wellman GC. Tonic regulation of middle meningeal artery diameter by ATP-sensitive potassium channels. J Cerebr Blood F Met 2017; 1: 271678X17749392.Google Scholar
  16. 16.
    Kubasch ML, Kubasch AS, Torres Pacheco J, et al. Laser doppler assessment of vasomotor axon reflex responsiveness to evaluate neurovascular function. Front Neurol 2017; 8: 370.Google Scholar
  17. 17.
    Tabrizchi R, Pvugsley MK. Methods of blood flow measurement in the arterial circulatory system. J Pharmacol Toxicol Methods 2000; 44(2): 375–384.Google Scholar
  18. 18.
    Bianchi B, Smith PA, Abriel H. The ion channel TRPM4 in murine experimental autoimmune encephalomyelitis and in a model of glutamate-induced neuronal degeneration. Mol Brain 2018; 11(1): 41.Google Scholar
  19. 19.
    Ma Z, Björklund A, Islam MS. A TRPM4 inhibitor 9-Phenanthrol inhibits glucose-and glucagon-like peptide 1-induced insulin secretion from rat islets of langerhans. J Diabetes Res 2017; 2017: 5131785.Google Scholar
  20. 20.
    Mironov SL. Calmodulin and CaMKII mediate emergent bursting activity in the brainstem respiratory network (preBötzinger complex). J Physiol 2013; 591(7): 1613–1630.Google Scholar
  21. 21.
    Wang C, Naruse K, Takahashi K. Role of the TRPM4 channel in cardiovascular physiology and pathophysiology. Cells 2018; 7(6): 62.Google Scholar
  22. 22.
    Liu H, El Zein L, Kruse M, et al. Gain-of-function mutations in TRPM4 cause autosomal dominant isolated cardiac conduction disease. Circ Cardiovasc Genet 2010; 3(4): 374–385.Google Scholar
  23. 23.
    Tian J, An XJ, Fu MY. Transient receptor potential melastatin 4 cation channel in pediatric heart block. Eur Rev Med Pharmacol Sci 2017; 21(4 Suppl): 79–84.Google Scholar
  24. 24.
    Palomares SM, Cipolla MJ. Myogenic tone as a therapeutic target for ischemic stroke. Curr Vasc Pharmacol 2014; 12(6): 788–800.Google Scholar
  25. 25.
    Ter Laan M, Van Dijk JM, Elting JW, Staal MJ, Absalom AR. Sympathetic regulation of cerebral blood flow in humans: a review. Br J Anaesth 2013; 111(3): 361–367.Google Scholar
  26. 26.
    Gonzales AL, Garcia ZI, Amberg GC, Earley S. Pharmacological inhibition of TRPM4 hyperpolarizes vascular smooth muscle. Am J Physiol Cell Physiol 2010; 299(5): C1195–1202.Google Scholar
  27. 27.
    An SJ, Kim TJ, Yoon BW. Epidemiology, risk factors, and clinical features of intracerebral hemorrhage: an update. J Stroke 2017; 19(1): 3–10.Google Scholar
  28. 28.
    Earley S. TRPM4 channels in smooth muscle function. Pflugers Arch 2013; 465(9): 1223–1231.Google Scholar
  29. 29.
    Earley S, Straub SV, Brayden JE. Protein kinase C regulates vascular myogenic tone through activation of TRPM4. Am J Physiol Heart Circ Physiol 2007; 292(6): H2613–2622.Google Scholar
  30. 30.
    Wang F, Wang Y, Xiao XF, Wang HZ, Sun T, Yu HL. Effect of arterial and venous subarachnoid hemorrhage on voltage-dependent calcium channel currents of cerebral artery smooth muscle cells. Chin J Cerebrovac Dis 2015; 12(2): 78–82.Google Scholar
  31. 31.
    Tsouka V, Markou T, Lazou A. Differential effect of ischemic and pharmacological preconditioning on PKC isoform translocation in adult rat cardiac myocytes. Cell Physiol Biochem 2002; 12(5–6): 315–324.Google Scholar
  32. 32.
    Zeng C, Tian F, Xiao B. TRPC channels: prominent candidates of underlying mechanism in neuropsychiatric diseases. Mol Neurobiol 2016; 53(1): 631–647.Google Scholar

Copyright information

© The American Society for Experimental NeuroTherapeutics, Inc. 2019

Authors and Affiliations

  • Yi Gong
    • 1
    • 2
    • 3
  • Ming-yue Du
    • 1
  • Hua-lin Yu
    • 1
  • Zhi-yong Yang
    • 1
  • Yu-jin Li
    • 4
  • Lei Zhou
    • 5
  • Rong Mei
    • 6
  • Li Yang
    • 7
  • Fei Wang
    • 1
    • 2
    Email author
  1. 1.Department of NeurosurgeryThe First Affiliated Hospital of Kunming Medical UniversityKunmingChina
  2. 2.Yunnan Key Laboratory of Laboratory MedicineKunmingChina
  3. 3.Department of NeurosurgeryThe Third People’s Hospital of Yunnan ProvinceKunmingChina
  4. 4.Department of AnesthesiologyThe First People’s Hospital of Yunnan ProvinceKunmingChina
  5. 5.The Key Laboratory of Stem Cell and Regenerative Medicine of Yunnan Province, Institute of Molecular and Clinical MedicineKunming Medical UniversityKunmingChina
  6. 6.Department of NeurologyThe First People’s Hospital of Yunnan ProvinceKunmingChina
  7. 7.Department of Anatomy, Histology and EmbryologyKunming Medical UniversityKunmingChina

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