Abstract

Although spatially restricted Ca2+ release from the endoplasmic reticulum (ER) through intracellular Ca2+ channels plays important roles in various neuronal activities, the accurate distribution and dynamics of ER in the dendrite of living neurons still remain unknown. To elucidate these, we expressed fluorescent protein-tagged ER proteins in cultured mouse hippocampal neurons, and monitored their movements using time-lapse microscopy. We report here that a sub-compartment of ER forms in relatively large vesicles that are capable, similarly to the reticular ER, of taking up and releasing Ca2+. The vesicular sub-compartment of ER moved rapidly along the dendrites in both anterograde and retrograde directions at a velocity of 0.2-0.3 microm/second. Depletion of microtubules, overexpression of dominant-negative kinesin and kinesin depletion by antisense DNA reduced the number and velocity of the moving vesicles, suggesting that kinesin may drive the transport of the vesicular sub-compartment of ER along microtubules in the dendrite. Rapid transport of the Ca2+-releasable sub-compartment of ER might contribute to rapid supply of fresh ER proteins to the distal part of the dendrite, or to the spatial regulation of intracellular Ca2+ signaling.

Highlights

  • Endoplasmic reticulum (ER) is an organelle that is responsible for the storage and release of Ca2+ in eukaryotic cells

  • We examined the distribution and dynamics of ER labeled with fluorescent proteins in the dendrite of cultured hippocampal neurons

  • Typical ER membrane proteins, a Ca2+ pump sarcoplasmic/endoplasmic reticulum calcium-ATPase 2a (SERCA2a) and a Ca2+-release channel Ins(1,4,5)P3R type1 [Ins(1,4,5)P3R1], both of which were tagged with green fluorescent protein (GFP) [GFP-SERCA2a and GFPIns(1,4,5)P3R1, respectively], were used to label ER membrane

Read more

Summary

Introduction

Endoplasmic reticulum (ER) is an organelle that is responsible for the storage and release of Ca2+ in eukaryotic cells. Inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]induced Ca2+ release (IICR) from intracellular calcium stores through Ins(1,4,5)P3 receptors [Ins(1,4,5)P3Rs] (Berridge, 1993) has been shown to be involved in depotentiation, suppression and input specificity of LTP, and in the induction of LTD in hippocampal neurons (Fujii et al, 2000; Nishiyama et al, 2000). Previous morphological and biochemical studies showing heterogeneous distribution of Ca2+ channels on ER and major ER resident proteins in Purkinje cells (Villa et al, 1991; Volpe et al, 1991; Takei et al, 1992) support this idea. The distribution of ER membrane in living neurons is little understood

Methods
Results
Discussion
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call