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De novo expression of Trpm4 initiates secondary hemorrhage in spinal cord injury

Abstract

The role of transient receptor potential M4 (Trpm4), an unusual member of the Trp family of ion channels, is poorly understood. Using rodent models of spinal cord injury, we studied involvement of Trpm4 in the progressive expansion of secondary hemorrhage associated with capillary fragmentation, the most destructive mechanism of secondary injury in the central nervous system. Trpm4 mRNA and protein were abundantly upregulated in capillaries preceding their fragmentation and formation of petechial hemorrhages. Trpm4 expression in vitro rendered COS-7 cells highly susceptible to oncotic swelling and oncotic death following ATP depletion. After spinal cord injury, in vivo gene suppression in rats treated with Trpm4 antisense or in Trpm4−/− mice preserved capillary structural integrity, eliminated secondary hemorrhage, yielded a threefold to fivefold reduction in lesion volume and produced a substantial improvement in neurological function. To our knowledge, this is the first example of a Trp channel that must undergo de novo expression for manifestation of central nervous system pathology.

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Figure 1: Trpm4 is upregulated in capillaries after SCI.
Figure 2: Antisense oligodeoxynucleotide downregulates Trpm4 expression and reduces secondary hemorrhage and capillary fragmentation after SCI.
Figure 3: Cells expressing Trpm4 are highly susceptible to ATP-depletion—induced oncotic cell death.
Figure 4: Trpm4-AS improves neurobehavioral performance and reduces lesion volume in rats after SCI.
Figure 5: Trpm4−/− reduces secondary hemorrhage and improves neurobehavioral performance in mice after SCI.

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Acknowledgements

This work was supported by grants to J.M.S. from the US National Heart, Lung, and Blood Institute (HL082517), the US National Institute of Neurological Disorders and Stroke (NS048260), the US Department of Veterans Affairs and the Christopher and Dana Reeve Foundation; to V.G. from the US National Institute of Neurological Disorders and Stroke (NS061934); and to M.F. and V.F. from the Deutsche Forschungsgemeinschaft. R.V. is supported by the FWO Vlaanderen. The mTrpm4-GFP plasmid was kindly provided by E.R. Liman (University of Southern California).

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Authors

Contributions

V.G. and J.M.S. conceived of and directed the study, participated in neurobehavioral assessments, analyzed the data and wrote the manuscript; S.K.W., Z.G. and Z.C. performed molecular and cell death experiments; O.T. performed rodent surgeries; S.I. performed immunolabeling and in situ hybridization; A.I. performed patch clamp experiments; R.V., M.F., V.F. and B.N. developed the Trpm4−/− mice and contributed to early drafts of the manuscript.

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Correspondence to J Marc Simard.

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Competing interests

J.M.S. and V.G. have applied for a US patent entitled "Therapeutic targeting of Trpm4 in CNS pathophysiology" (application number 60/889,065). S.K.W., R.V., O.T., S.I., A.I., Z.G., Z.C., V.F., B.N. and M.F. declare no competing financial interests.

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Gerzanich, V., Woo, S., Vennekens, R. et al. De novo expression of Trpm4 initiates secondary hemorrhage in spinal cord injury. Nat Med 15, 185–191 (2009). https://doi.org/10.1038/nm.1899

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