Rapid 3D Enhanced Resolution Microscopy Reveals Diversity in Dendritic Spinule Dynamics, Regulation, and Function

Neuron. 2020 Aug 5;107(3):522-537.e6. doi: 10.1016/j.neuron.2020.04.025. Epub 2020 May 27.


Dendritic spinules are thin protrusions, formed by neuronal spines, not adequately resolved by diffraction-limited light microscopy, which has limited our understanding of their behavior. Here we performed rapid structured illumination microscopy and enhanced resolution confocal microscopy to study spatiotemporal spinule dynamics in cortical pyramidal neurons. Spinules recurred at the same locations on mushroom spine heads. Most were short-lived, dynamic, exploratory, and originated near simple PSDs, whereas a subset was long-lived, elongated, and associated with complex PSDs. These subtypes were differentially regulated by Ca2+ transients. Furthermore, the postsynaptic Rac1-GEF kalirin-7 regulated spinule formation, elongation, and recurrence. Long-lived spinules often contained PSD fragments, contacted distal presynaptic terminals, and formed secondary synapses. NMDAR activation increased spinule number, length, and contact with distal presynaptic elements. Spinule subsets, dynamics, and recurrence were validated in cortical neurons of acute brain slices. Thus, we identified unique properties, regulatory mechanisms, and functions of spinule subtypes, supporting roles in neuronal connectivity.

Keywords: calcium transients; dendritic spine dynamics; intrabodies; kalirin-7; live imaging; multi-synaptic spines; postsynaptic density; structured illumination microscopy; synaptic plasticity; synaptic spinules.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Animals
  • Calcium / metabolism
  • Cerebral Cortex / cytology
  • Dendritic Spines / metabolism
  • Dendritic Spines / physiology
  • Dendritic Spines / ultrastructure*
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Imaging, Three-Dimensional
  • Mice
  • Microscopy, Confocal
  • Post-Synaptic Density / physiology
  • Post-Synaptic Density / ultrastructure*
  • Pyramidal Cells / physiology
  • Pyramidal Cells / ultrastructure*
  • Receptors, N-Methyl-D-Aspartate / agonists
  • Spatio-Temporal Analysis
  • Synapses / physiology
  • Synapses / ultrastructure*


  • Guanine Nucleotide Exchange Factors
  • KALRN protein, mouse
  • Receptors, N-Methyl-D-Aspartate
  • Calcium