Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent
Abstract One of the mechanisms of calcium signalling in neurons is store-operated calcium entry (SOCE), which is activated when the calcium concentration in the smooth endoplasmic reticulum (ER) decreases and its protein-calcium sensor STIM (stromal interacting molecule) relocate to the endoplasmic...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-024-85024-5 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841544569120882688 |
|---|---|
| author | Anastasiya Rakovskaya Ekaterina Volkova Ilya Bezprozvanny Ekaterina Pchitskaya |
| author_facet | Anastasiya Rakovskaya Ekaterina Volkova Ilya Bezprozvanny Ekaterina Pchitskaya |
| author_sort | Anastasiya Rakovskaya |
| collection | DOAJ |
| description | Abstract One of the mechanisms of calcium signalling in neurons is store-operated calcium entry (SOCE), which is activated when the calcium concentration in the smooth endoplasmic reticulum (ER) decreases and its protein-calcium sensor STIM (stromal interacting molecule) relocate to the endoplasmic reticulum and plasma membrane junctions, forms clusters and induces calcium entry. In electrically non-excitable cells, STIM1 is coupled with the positive end of a tubulin microtubule through interaction with EB1 (end-binding) protein, which controls its oligomerization, SOCE and participates in ER movement. STIM2 homologue, which is specific for mature hippocampal dendritic spines, is known to interact with EB3 protein, however, not much is known about the role of this interaction in STIM2 clustering or ER trafficking in neurons. Intriguingly, in neurons, reducing the expression of EB3 protein or disrupting the interaction of STIM2 protein with EB proteins results in decreased SOCE, in contrast to experiments with STIM1 in non-excitable cells. In this study, these two homologues are compared side-by-side in HEK-293T, and it is shown for the first time that their clustering and SOCE is oppositely regulated by dynamic tubulin microtubules. In particular, for STIM2, the interaction with dynamic microtubule cytoskeleton is required for clustering and is shown to potentiate SOCE, while for STIM1 this interaction restricts clustering, resulting in SOCE decrease. After store depletion in primary hippocampal neurons, the wild type STIM2 is redistributed from the necks to the heads of dendritic spines, while the STIM2 variant with a mutation that disrupts the interaction with EB proteins is excluded from dendritic spines. In addition, overexpression of the mutant variant leads to ER reorganization in neuronal soma and reduction of ER presence in spines. It also leads to a reduction in the number of spines containing the spine apparatus formed by ER cisternae, as well as a reduction in dendritic spines SOCE. These effects are opposite of those detected during overexpression of the wild type STIM2. Considered together, these findings underline the important role of dynamic microtubules in regulation of neuronal SOCE and ER morphology. |
| format | Article |
| id | doaj-art-21f068bfc08e44f2b51b0c96042214c2 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-21f068bfc08e44f2b51b0c96042214c22025-01-12T12:24:13ZengNature PortfolioScientific Reports2045-23222025-01-0115111410.1038/s41598-024-85024-5Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependentAnastasiya Rakovskaya0Ekaterina Volkova1Ilya Bezprozvanny2Ekaterina Pchitskaya3Laboratory of Biomedical Imaging and Data Analysis, Institute of Biomedical Systems and Biotechnology, Peter the Great St. Petersburg Polytechnic UniversityLaboratory of Biomedical Imaging and Data Analysis, Institute of Biomedical Systems and Biotechnology, Peter the Great St. Petersburg Polytechnic UniversityLaboratory of Molecular Neurodegeneration, Institute of Biomedical Systems and Biotechnology, Peter the Great St. Petersburg Polytechnic UniversityLaboratory of Biomedical Imaging and Data Analysis, Institute of Biomedical Systems and Biotechnology, Peter the Great St. Petersburg Polytechnic UniversityAbstract One of the mechanisms of calcium signalling in neurons is store-operated calcium entry (SOCE), which is activated when the calcium concentration in the smooth endoplasmic reticulum (ER) decreases and its protein-calcium sensor STIM (stromal interacting molecule) relocate to the endoplasmic reticulum and plasma membrane junctions, forms clusters and induces calcium entry. In electrically non-excitable cells, STIM1 is coupled with the positive end of a tubulin microtubule through interaction with EB1 (end-binding) protein, which controls its oligomerization, SOCE and participates in ER movement. STIM2 homologue, which is specific for mature hippocampal dendritic spines, is known to interact with EB3 protein, however, not much is known about the role of this interaction in STIM2 clustering or ER trafficking in neurons. Intriguingly, in neurons, reducing the expression of EB3 protein or disrupting the interaction of STIM2 protein with EB proteins results in decreased SOCE, in contrast to experiments with STIM1 in non-excitable cells. In this study, these two homologues are compared side-by-side in HEK-293T, and it is shown for the first time that their clustering and SOCE is oppositely regulated by dynamic tubulin microtubules. In particular, for STIM2, the interaction with dynamic microtubule cytoskeleton is required for clustering and is shown to potentiate SOCE, while for STIM1 this interaction restricts clustering, resulting in SOCE decrease. After store depletion in primary hippocampal neurons, the wild type STIM2 is redistributed from the necks to the heads of dendritic spines, while the STIM2 variant with a mutation that disrupts the interaction with EB proteins is excluded from dendritic spines. In addition, overexpression of the mutant variant leads to ER reorganization in neuronal soma and reduction of ER presence in spines. It also leads to a reduction in the number of spines containing the spine apparatus formed by ER cisternae, as well as a reduction in dendritic spines SOCE. These effects are opposite of those detected during overexpression of the wild type STIM2. Considered together, these findings underline the important role of dynamic microtubules in regulation of neuronal SOCE and ER morphology.https://doi.org/10.1038/s41598-024-85024-5Dendritic spineSpine apparatusEndoplasmic reticulumStore-operated calcium entryEnd-binding proteinsDynamic microtubules |
| spellingShingle | Anastasiya Rakovskaya Ekaterina Volkova Ilya Bezprozvanny Ekaterina Pchitskaya Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent Scientific Reports Dendritic spine Spine apparatus Endoplasmic reticulum Store-operated calcium entry End-binding proteins Dynamic microtubules |
| title | Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent |
| title_full | Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent |
| title_fullStr | Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent |
| title_full_unstemmed | Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent |
| title_short | Hippocampal dendritic spines store-operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent |
| title_sort | hippocampal dendritic spines store operated calcium entry and endoplasmic reticulum content is dynamic microtubule dependent |
| topic | Dendritic spine Spine apparatus Endoplasmic reticulum Store-operated calcium entry End-binding proteins Dynamic microtubules |
| url | https://doi.org/10.1038/s41598-024-85024-5 |
| work_keys_str_mv | AT anastasiyarakovskaya hippocampaldendriticspinesstoreoperatedcalciumentryandendoplasmicreticulumcontentisdynamicmicrotubuledependent AT ekaterinavolkova hippocampaldendriticspinesstoreoperatedcalciumentryandendoplasmicreticulumcontentisdynamicmicrotubuledependent AT ilyabezprozvanny hippocampaldendriticspinesstoreoperatedcalciumentryandendoplasmicreticulumcontentisdynamicmicrotubuledependent AT ekaterinapchitskaya hippocampaldendriticspinesstoreoperatedcalciumentryandendoplasmicreticulumcontentisdynamicmicrotubuledependent |