Abstract
The establishment of polarity is an essential process in early neuronal development. Cdc42, a GTPase of the Rho family, is a key regulator of cytoskeletal dynamics and neuronal polarity. However, the mechanisms underlying the action of cdc42 in regulating axonogenesis have not been elucidated. Here, we expressed wild-type cdc42, a constitutively active cdc42 mutant (cdc42F28L) and a dominant negative cdc42 mutant (cdc42N17), respectively, in the primary hippocampal neurons to alter the activity of cdc42. We found that cdc42 activities were paralleled with the capacities to promote axonogenesis in the cultured neurons. Cdc42 also enhanced microtubule stability in the cultured neurons. Pharmacologically stabilizing microtubules significantly abrogated the defective axonogenesis induced by cdc42 inhibition. Moreover, cdc42 promoted the dephosphorylation of collapsing response mediator protein-2 (CRMP-2) at Thr514 by increasing GSK-3β phosphorylation at Ser9 in the cultured neurons. These findings suggest that cdc42 may facilitate axonogenesis by promoting microtubule stabilization in rat primary hippocampal neurons.
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Data Availability
The raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher.
Abbreviations
- CRMP-2:
-
Collapsing response mediator protein-2
- cdc42wt:
-
Wild-type cdc42
- mAb:
-
Monoclonal antibodies
- pAb:
-
Polyclonal antibodies
- SD:
-
Sprague–Dawley
- PVDF:
-
Polyvinylidene difluoride
- ECL:
-
Enhanced chemiluminescence
- A/T:
-
Acetylated to tyrosinated α-tubulin
- RGCs:
-
Xenopus retinal ganglion cells
- GSK-3β:
-
Glycogen synthase kinase-3β
- aPKC:
-
Atypical protein kinase C
- NA:
-
No axon
- SA:
-
A single axon
- MA:
-
Multiple axons
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Funding
This work is supported in part by grants from the National Natural Science Foundation of China (81572222 and 30900725) and Guangdong Provincial Natural Science Foundation (2014A030313333 and 8151051501000005).
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AL and HZ contributed equally to this work; HW designed research; AL, HZ, YC, FY, ZYL, and ZLL performed research; AL, HZ, WD, LZ, and HW analyzed data; and HW wrote the manuscript. All authors approved the final manuscript.
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The experimental protocol was established according to the Guidelines for the Care and Use of Laboratory Animals of the National Institutes of Health and approved by the Animal Care and Use Committee of Southern Medical University.
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Li, A., Zhu, HM., Chen, Y. et al. Cdc42 Facilitates Axonogenesis by Enhancing Microtubule Stabilization in Primary Hippocampal Neurons. Cell Mol Neurobiol 41, 1599–1610 (2021). https://doi.org/10.1007/s10571-021-01051-0
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DOI: https://doi.org/10.1007/s10571-021-01051-0