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Vaidya, R. M.

Publications and source records attributed to Vaidya, R. M..

3 recordsLinked to original sources

Nanoscale organization is changed in native, surface AMPARs by mouse brain region and tauopathy

The distribution of synaptic and extra-synaptic AMPA receptors (AMPARs) on neuronal plasma membranes is correlated with learning and memory. Although AMPAR organization has been extensively studied in neuronal cultures, its native cell-surface distribution in intact adult brain tissue across distinct brain regions and in neurodegenerative pathology remains poorly understood. Here, we combine a selective small-molecule labeling strategy with two-color 3D super-resolution dSTORM imaging to map native surface AMPAR organization at the nanoscale in 30 micron thick mouse brain slices. We find that wild-type mice exhibit marked regional differences in AMPAR organization, with the CA1 hippocampus containing a substantially larger extrasynaptic AMPAR pool than the nearby motor and somatosensory cortex. In the PS19 tauopathy mouse model, at an age preceding overt neurodegeneration, AMPAR organization is selectively disrupted in the hippocampus but largely preserved in the cortex. Specifically, we observe depletion of the extrasynaptic receptor pool together with reduced synaptic nanodomain organization, revealing early molecular-scale synaptic remodeling associated with tau pathology. These findings provide direct structural insight into region- and disease-dependent AMPAR organization in intact adult brain tissue and establish a broadly applicable framework for nanoscale investigation of synaptic receptor architecture in health and neurological disease.

neuroscience↗

Automatic dendritic spine segmentation in widefield fluorescence images reveal synaptic nanostructures distribution with super-resolution imaging

Relating dendritic spine morphology to synaptic organization in brain tissue is essential for understanding excitatory synaptic transmission and plasticity. Single-molecule localization microscopy (SMLM) offers the spatial precision needed to study the synaptic protein distribution at the nanoscale. However, the widefield setup required for SMLM produces diffraction-limited images with poor contrast and resolution in thick brain slices (> 30 m), making accurate segmentation of dendritic spines challenging. To overcome this challenge, we developed an automated 3D segmentation approach tailored to this condition by combining two existing machine-learning models. We integrated this strategy with SMLM-based localization of synaptic proteins to map post-synaptic protein PSD-95 within spines at nanoscale resolution. This framework, named ISEPLA (Integrated Spine Extraction and Protein Localization Analysis), revealed a hierarchical organization of synaptic proteins: spines contain multiple nanomodules, each composed of smaller nanodomains. Larger spines contain more nanomodules, and larger nanomodules comprise more nanodomains. Therefore, our method enables precise morphological and molecular analysis under physiologically relevant imaging conditions, providing new insights into the synaptic organization in spines.

neuroscience↗

Asymmetric Tug-of-War leads to Cooperative Transport of a Cargo by Multiple Kinesins

How cargoes move within a crowded cell--over long distances and at speeds that are nearly the same as when moving on an unimpeded pathway--has long been mysterious. Through an in vitro gliding assay, which involves measuring nanometer displacement and piconewtons of force, we have evidence that when kinesins, a cytoplasmic molecular motor, operate in small groups, from 2-10, they can communicate among themselves through an asymmetric tug-of-war by inducing tension (up to 4 pN) on the cargo. Surprisingly, the primary role of approximately one-third of kinesins is to develop tension, which instantaneously slows forward motion but helps increase cargo run length. These hindering kinesins fall off rapidly when experiencing a forward tug. Occasionally, they may be ripped off from their anchors by other driving kinesins working in tandem. Furthermore, with roadblocks on the microtubule, multiple kinesins cooperate to overcome impediments. Hence, kinesin may employ an asymmetric tug-of-war and a cooperative motion to navigate through cellular environment.

biophysics↗