Biology subjects

Prato, V.

Publications and source records attributed to Prato, V..

4 recordsLinked to original sources

bioRxiv2026

Junctional Nanodomains Organised by Extended Synaptotagmin-1 Underlie Inflammatory Signalling in Sensory Neurons

Endoplasmic reticulum-plasma membrane (ER-PM) junctions coordinate Ca2+ signalling and nociceptor excitability during inflammation, but the molecular scaffold governing assembly of signalling complexes remains unclear. Here, we identify Extended Synaptotagmin-1 (Esyt1) as a key, dynamic regulator of inflammatory pain signalling. In nociceptive DRG neurons, Esyt1 is co-expressed with ANO1 and IP3R1. Time-resolved proximity ligation assays and super-resolution microscopy showed that bradykinin rapidly increased ANO1-Esyt1 nanoscale proximity, while IP3R1-Esyt1 interactions were dynamically reorganised during ANO1-IP3R1 complex formation. Esyt1 knockdown reduced bradykinin-evoked ANO1 activity, impaired store-operated Ca2+ entry (SOCE), decreased STIM1-Orai1 complex formation, and disrupted ER Ca2+ store refilling. Simultaneous imaging of Ca2+ dynamics and ANO1 activity demonstrated functional coupling between SOCE and ANO1 that required Esyt1. Furthermore, in-vivo Esyt1-knockdown significantly reduced bradykinin-induced nocifensive behaviour. Together, these findings identify Esyt1 as a Ca2+-responsive scaffold that organises ER-PM signalling nanodomains and promotes inflammatory pain signalling.

neuroscience↗

bioRxiv2026

ESCRT-I Inhibition Protects against NMDAR Hypofunction and Restores Synaptic Homeostasis in a Cellular Model of Schizophrenia

N-methyl-D-aspartate receptor (NMDAR) hypofunction is a central pathophysiological mechanism in schizophrenia, yet direct NMDAR potentiation has shown limited clinical benefit, potentially because it fails to address deficits in receptor trafficking and surface stability. The endosomal sorting complexes required for transport (ESCRT) machinery governs the lysosomal fate of internalised membrane proteins, but its role in synaptic receptor homeostasis during glutamatergic dysfunction remains unclear. Phencyclidine (PCP), a non-competitive NMDAR antagonist, is widely used to model NMDAR hypofunction and schizophrenia-like phenotypes. Here, we show that sub-chronic PCP exposure induced persistent NMDAR hypofunction, impaired GABAergic transmission, and collapsed of excitation/inhibition (E/I) balance in primary hippocampal neurons. Genetic inhibition of the ESCRT-I component TSG101 prevented these deficits, enabled functional recovery following PCP washout, and increased surface expression of functional NMDARs and GABAA receptors without altering receptor pharmacology or biophysical properties. At the network level, TSG101 knockdown restored inhibitory tone, normalised excitatory activity, and stabilised E/I balance. Moreover, TSG101 knockdown rescued PCP-induced reductions in PSD-95 and BDNF, restored ERK1/2 signalling, and re-established activity-dependent nuclear translocation of Fos-like (Fos-L), consistent with reactivation of transcriptional plasticity. These findings identify ESCRT-I as a regulator of synaptic stability, positioning endosomal sorting pathways as a potential therapeutic target for schizophrenia.

neuroscience↗

bioRxiv2023

Peripheral gating of pain by glial endozepine

We report that diazepam binding inhibitor (DBI) is a glial messenger mediating satellite glia-sensory neuron crosstalk in the dorsal root ganglion (DRG). DBI is highly and specifically expressed in satellite glia cells (SGCs) of mice, rat and human, but not in sensory neurons or other DRG-resident cells. Knockdown of DBI results in a robust mechanical hypersensitivity without significant effects on other sensory modalities. In vivo overexpression of DBI in SGCs reduces sensitivity to mechanical stimulation and alleviates mechanical allodynia in neuropathic and inflammatory pain models. We further show that DBI acts as a partial agonist and positive allosteric modulator at the neuronal GABAA receptors, particularly strongly effecting those with a high-affinity benzodiazepine binding site. Such receptors are selectively expressed by a subpopulation of mechanosensitive DRG neurons and these are also more enwrapped with DBI-expressing glia, as compared to other DRG neurons, suggesting a mechanism for specific effect of DBI on mechanosensation. These findings identified a new, peripheral neuron-glia communication mechanism modulating pain signalling, which can be targeted therapeutically.

neuroscience↗

bioRxiv2022

The molecular mechanism and physiological role of silent nociceptor activation

Silent nociceptors are sensory afferents that are insensitive to noxious mechanical stimuli under normal conditions but become sensitized to such stimuli during inflammation. Using RNA-sequencing and quantitative RT-PCR we demonstrate that inflammation selectively upregulates the expression of the transmembrane protein TMEM100 in silent nociceptors and electrophysiology revealed that over-expression of TMEM100 is required and sufficient to un-silence silent nociceptors. Moreover, we show that mice lacking TMEM100 do not develop secondary allodynia - i.e. pain hypersensitivity that spreads beyond the site of inflammation - in a mouse model of knee joint inflammation and that AAV-mediated overexpression of TMEM100 in articular afferents in the absence of inflammation is sufficient to induce allodynia in remote skin regions without causing knee joint pain. Thus, our work identifies TMEM100 as a key regulator of silent nociceptor un-silencing and reveals a physiological role for this hitherto enigmatic afferent subclass in triggering spatially remote secondary allodynia during inflammation.

neuroscience↗