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Ras-Related C3 Botulinum Toxin Substrate 1 Promotes Axonal Regeneration after Stroke in Mice

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Abstract

Neurite plasticity is a critical aspect of brain functional recovery after stroke. Emerging data suggest that Ras-related C3 botulinum toxin substrate 1 (Rac1) plays a central role in axonal regeneration in the injured brain, specifically by stimulating neuronal intrinsic growth and counteracting the growth inhibitory signaling that leads to growth cone collapse. Therefore, we investigated the functional role of Rac1 in axonal regeneration after stroke.

Delayed treatment with a specific Rac1 inhibitor, NSC 23766, worsened functional recovery, which was assessed by the pellet reaching test from day 14 to day 28 after stroke. It additionally reduced axonal density in the peri-infarct zone, assessed 28 days after stroke, with no effect on brain cavity size or on the number of newly formed cells. Accordingly, Rac1 overexpression using lentivirus promoted axonal regeneration and functional recovery after stroke from day 14 to day 28. Rac1 inhibition led to inactivation of pro-regenerative molecules, including mitogen-activated protein kinase kinase (p-MEK)1/2, LIM domain kinase (LIMK)1, and extracellular signal-regulated kinase (p-ERK)1/2 at 14 days after stroke. Inhibition of Rac1 reduced axonal length and number in cultured primary mouse cortical neurons using microfluidic chambers after oxygen-glucose deprivation (OGD) without affecting cell viability. In contrast, inhibition of Rac1 increased levels of glial fibrillary acidic protein, an extrinsic inhibitory signal for axonal growth, after stroke in vivo and in primary astrocytes after OGD.

In conclusion, Rac1 signaling enhances axonal regeneration and improve post-stroke functional recovery in experimental models of stroke.

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Acknowledgements

This project was generously supported by National Institutes of Health grants R01 NS099628 (J.Li) and R01 NS078446 (J.Li). This project was generously supported by National Institutes of Health grants R01 NS099628 (J.Li) and R01 NS078446 (J.Li).

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Authors and Affiliations

  1. Department of Neurology, University of Texas Health Science Center, Houston, TX, USA

    Lin Liu, Hui Yuan, Yanhua Yi, Edward C. Koellhoffer, Yashasvee Munshi, Fan Bu, Louise D. McCullough & Jun Li

  2. Department of Neurology, Henry Ford Hospital, Detroit, MI, USA

    Yi Zhang & Zhenggang Zhang

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  1. Lin Liu
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Correspondence to Jun Li.

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All procedures were approved by the Center for Laboratory Animal Care of the University of Connecticut Health Center and University of Texas Health Science Center and performed following the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Liu, L., Yuan, H., Yi, Y. et al. Ras-Related C3 Botulinum Toxin Substrate 1 Promotes Axonal Regeneration after Stroke in Mice. Transl. Stroke Res. 9, 506–514 (2018). https://doi.org/10.1007/s12975-018-0611-5

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