The non-mitotic role of HMMR in regulating the localization of TPX2 and the dynamics of microtubules in neurons

A functional nervous system is built upon the proper morphogenesis of neurons to establish the intricate connection between them. The microtubule cytoskeleton is known to play various essential roles in this morphogenetic process. While many microtubule-associated proteins (MAPs) have been demonstrated to participate in neuronal morphogenesis, the function of many more remains to be determined. This study focuses on a MAP called HMMR in mice, which was originally identified as a hyaluronan binding protein and later found to possess microtubule and centrosome binding capacity. HMMR exhibits high abundance on neuronal microtubules and altering the level of HMMR significantly affects the morphology of neurons. Instead of confining to the centrosome(s) like cells in mitosis, HMMR localizes to microtubules along axons and dendrites. Furthermore, transiently expressing HMMR enhances the stability of neuronal microtubules and increases the formation frequency of growing microtubules along the ..., Indirect immunofluorescence staining
 
Cells on coverslips were fixed with 3.7% formaldehyde for 15 min at 37 °C and then washed three times with PBS. Fixed cells were permeabilized with 0.25% triton X-100 in PBS for 5 min at room temperature or extracted in -20 °C methanol for 10 min. For experiments that required cytosolic pre-extraction, cells on coverslips were permeabilized in 0.1% triton X-100 in PIPES buffer (0.1 M PIPES pH 6.9, 1 mM MgCl2, and 1 mM EGTA) for 15 sec, washed once with PIPES buffer, and fixed with 3.7% formaldehyde in PIPES buffer at 37 °C for 30 min and then washed with PBS three times. Cells were then blocked with 10% BSA in PBS for 30 min at 37 °C, incubated for 1 hour at 37 °C with different primary antibodies: GFP (1:100), HMMR (1:50), MAP2 (1:1000), SMI312 (1:1000), TPX2 (1:2000), acetylated-α-tubulin (1:1000), TUBB3 (1:2000), and TUJ1 (1:4000). After primary antibody incubation, cells were washed with PBS three times and incubated with AlexaFluor-conjugated ..., , # The non-mitotic role of HMMR in regulating the localization of TPX2 and the dynamics of microtubules in neurons

[https://doi.org/10.5061/dryad.cz8w9gjbz](https://doi.org/10.5061/dryad.cz8w9gjbz)

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