Venkatesan, S and Subramaniam, S and Rajeev, P and Chopra, Y and Jose, M and Nair, D (2020) Differential scaling of synaptic molecules within functional zones of an excitatory synapse during homeostatic plasticity. In: eNeuro, 7 (2). pp. 1-16.
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Abstract
Homeostatic scaling is a form of synaptic plasticity where individual synapses scale their strengths to compensate for global suppression or elevation of neuronal activity. This process can be studied by measuring miniature EPSP (mEPSP) amplitudes and frequencies following the regulation of activity in neuronal cultures. Here, we demonstrate a quantitative approach to characterize multiplicative synaptic scaling using immunolab-elling of hippocampal neuronal cultures treated with tetrodotoxin (TTX) or bicuculline to extract scaling factors for various synaptic proteins. This approach allowed us to directly examine the scaling of presynaptic and postsynaptic scaffolding molecules along with neurotransmitter receptors in primary cultures from mouse and rat hippocampal neurons. We show robust multiplicative scaling of synaptic scaffolding molecules namely, Shank2, PSD95, Bassoon, and AMPA receptor subunits and quantify their scaling factors. We use super-reso-lution microscopy to calculate scaling factors of surface expressed GluA2 within functional zones of the synapse and show that there is differential and correlated scaling of GluA2 levels within the spine, the postsynaptic density (PSD), and the perisynaptic regions. Our method opens a novel paradigm to quantify relative molecular changes of synaptic proteins within distinct subsynaptic compartments from a large number of synapses in response to alteration of neuronal activity, providing anatomic insights into the intricacies of vari-ability in strength of individual synapses. © 2020, Society for Neuroscience. All rights reserved.
| Item Type: | Journal Article |
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| Publication: | eNeuro |
| Publisher: | Society for Neuroscience |
| Additional Information: | The copyright for this article belongs to the Authors. |
| Keywords: | scaffold protein; tetrodotoxin; AMPA receptor; nerve protein; Shank2 protein, mouse, animal experiment; animal model; Article; comparative study; confocal microscopy; controlled study; correlational study; excitatory postsynaptic potential; gene; GluA2 gene; hemostasis; hippocampus; image analysis; immunocytochemistry; morphometry; nerve cell plasticity; nonhuman; postsynaptic density; priority journal; quantitative analysis; rat; synapse vesicle; animal; genetics; homeostasis; mouse; nerve cell; synapse, Animals; Homeostasis; Mice; Nerve Tissue Proteins; Neuronal Plasticity; Neurons; Rats; Receptors, AMPA; Synapses |
| Department/Centre: | Division of Biological Sciences > Centre for Neuroscience |
| Date Deposited: | 24 Jan 2023 09:29 |
| Last Modified: | 24 Jan 2023 09:29 |
| URI: | https://eprints.iisc.ac.in/id/eprint/79405 |
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