Biology subjects

Gazzola, M.

Publications and source records attributed to Gazzola, M..

9 recordsLinked to original sources

bioRxiv2026

ABISS: An Open-Source, Low-Cost Platform for Auditory and Visual Intrinsic Optical Signal Imaging

Defining the boundaries of functional cortical areas is increasingly important for targeted electro-physiology, optical imaging, viral delivery, and circuit manipulation. Intrinsic optical signal imaging (IOSI) provides a rapid and minimally invasive approach for mapping stimulus-evoked cortical activity, but its implementation often requires laboratory-specific combinations of stimulus-generation hardware, experiment-control software, synchronization devices, and data-acquisition systems. These requirements limit accessibility and hinder the use of IOSI as a routine functional mapping tool. Here, we present the Arduino-Based Intrinsic Stimulation System (ABISS), an open-source platform that integrates auditory and visual stimulus generation, trial timing, and image-acquisition triggering into a single programmable device. ABISS generates auditory tone stimuli, VGA-based visual stimuli, and tightly synchronized camera-trigger pulses without requiring a dedicated experiment-control system. Stimulus protocols are also fully modifiable in firmware. Performance was evaluated in auditory and visual cortices of mice. Engineering validation demonstrated accurate stimulus generation and synchronization between stimulus delivery and camera triggering over extended recording sessions. Biological validation showed that ABISS output results in auditory and visual intrinsic signal maps comparable to those obtained using highly specialized or commercial platforms. Together, these findings demonstrate the utility of a low-cost open-source platform for experimental control of intrinsic optical signal imaging. By reducing the technical and financial barriers associated with routine intrinsic optical imaging, ABISS facilitates broader adoption of functional cortical mapping as a tool for improved cortical localization in neuroscience experiments. Significance StatementFunctional cortical mapping is an increasingly important element of neuroscience experimental design as anatomical coordinates alone are often insufficient for defining cortical boundaries in individual animals. Intrinsic optical signal imaging provides an effective solution but traditionally requires specialized hardware, commercial stimulus-generation systems, and laboratory-specific synchronization workflows. We developed ABISS, an inexpensive, open-source platform that integrates auditory and visual stimulus generation with synchronized camera triggering in a single programmable device. ABISS produces functional cortical maps comparable to those obtainable with commercial or specialized systems while substantially reducing hardware complexity and cost. By making intrinsic optical signal imaging more accessible, ABISS lowers the practical barriers for routine functional mapping of the brain and promotes adoption of this important neuroscience technique.

neuroscience↗

bioRxiv2025

Living Neurosheets: Engineering readily deployable neural architectures

Inspired by the ability of brain structures to support cognitive processing, the need for reliable queryable neural models, and emerging biohybrid applications, we present Living Neurosheets: practical, malleable, modular, and scalable systems for realizing living neural architectures, bio-inspired or beyond evolutionary precedent. By innervating porous sheets, paper-like substrates are endowed with neural function while retaining exceptional versatility. Pre-programmed porosity anchors deposited neurons for reliable spatial control. Cutting, folding, and stacking allow intuitive design, modular composition, and batch fabrication (by the hundreds) of complex neural motifs that preserve geometry and function for months. Reversible integration with tissue-matched optoelectronics enables both sample reuse across hardware and hardware multiplexing across samples, for high-throughput, multimodal investigations. We illustrate these aspects across tissue topologies, gyri-inspired manifolds, and laminar cortical mimics, attaining control over entrainment, synchronization, propagation, suppression, and augmentation dynamics--key features of in vivo neural activity. Finally, via cross-instrument portability, we fuse spatial transcriptomics and electrophysiology, linking gene programs to emergent dynamics.

neuroscience↗

bioRxiv2024

Alternative splicing of SORBS1 affects neuromuscular junction formation and stability in myotonic dystrophy type 1

Myotonic dystrophy type 1 (DM1) is a multisystemic neuromuscular disease characterized by a CTG repeat expansion in the 3" untranslated region of the gene coding for the dystrophia myotonica protein kinase (DMPK). Presence of expanded CTG repeats in DMPK-mRNAs leads to the sequestration of RNA binding factors such as the Muscleblind like (MBNL) proteins resulting in widespread splicing defects contributing to progressive muscle weakness and myotonia. Here, we show that abnormal splicing of SORBS1 exon 25 found in skeletal muscle of myotonic dystrophy type 1 patients is a critical contributor to neuromuscular junction (NMJ) formation and maintenance. Forced exclusion of SORBS1 exon 25 in mice results in NMJ degeneration with marked denervation and postsynaptic destabilization. In zebrafish, misregulation of sorbs1 exon 25 results in reduced motor function and abnormal AChR cluster morphology. Finally, we observed that forcing SORBS1 exon 25 exclusion in hiPSC-derived skeletal muscle cells reduces the formation of large AChR clusters upon agrin stimulation. Thus, our study identifies MBNL regulated SORBS1 alternative splicing during skeletal muscle development as a critical event for NMJ formation and maintenance. The aberrant splicing of SORBS1 exon 25 in DM1 expands our understanding of how splicing dysregulation compromises neuromuscular system communication, shedding light on the broader impact of mRNA splicing regulation on NMJ integrity.

molecular biology↗

bioRxiv2024

Airway smooth muscle tone curbs hyperresponsiveness in experimental asthma

Background & ObjectivesA sustained contraction of airway smooth muscle, hereinafter called tone, increases the response to methacholine in healthy mice and humans. However, the effect of tone in the context of an active inflammation remains to be investigated. The objective of the present study was to test the effect of tone on the in vivo response to methacholine in mice during an active inflammatory phase of experimental asthma. MethodsMale BALB/c mice were exposed once-daily to either intranasal saline or house dust mite for 10 consecutive days to induce experimental asthma. They then underwent one of two methacholine challenges 24 h after the last exposure. While the same cumulative dose was administered in both challenges, one was preceded by a 20-min period of tone induced by nebulizing low doses of methacholine at 5-min intervals. Respiratory mechanics were monitored before and throughout the methacholine challenge by oscillometry. Bronchoalveolar lavages (BAL) and histology were also performed. ResultsBAL inflammation and histological alterations were consistent with experimental asthma. In accordance with previous studies, tone potentiated the response to methacholine in control mice, mainly by stiffening the lung periphery. The lung was even stiffer upon methacholine challenge during an active phase of inflammation in mice with experimental asthma, but this was not further potentiated by tone. In fact, in mice with experimental asthma, tone mitigated hyperresponsiveness by preventing further airway narrowing and, more importantly, small airway narrowing heterogeneity and closure. ConclusionDuring an active inflammatory phase of experimental asthma, tone protects against hyperresponsiveness. Take-home messageThe effect of airway smooth muscle tone on the methacholine response was investigated in mice with or without experimental asthma. While tone potentiated the methacholine response in control mice, it mitigated hyperresponsiveness in experimental asthma. These results unveiled a protective role of the airway smooth muscle in experimental asthma.

physiology↗

bioRxiv2024

Centrosome positioning independently of microtubule-based forces

The regulation of centrosome position is central to the establishment of polarized cell functions. This process is thought to rely on the generation of mechanical forces along microtubules, the balance of which determines centrosome position. By studying these forces in adherent cells in culture, we found that neither pushing nor pulling forces propagate along microtubules to the centrosome. Inhibiting dyneins or disassembling microtubules did not disrupt the maintenance of the centrosome at the center of the cell. In contrast, the actomyosin contractile network appears to be responsible for the generation of a centripetal flow that drives the centrosome towards the center of the cell independently of the microtubules. Furthermore, we found that centering of the centrosome depends more on the reorganization of cell shape around the centrosome than on an effective centrosome displacement throughout the cytoplasm. Interestingly, despite their lack of mechanical role, microtubules appear to direct this remodeling of cell shape. This revised view of centrosome positioning offers a new perspective for understanding the establishment of cell polarity.

cell biology↗

bioRxiv2023

`Mind in Vitro' platforms: Versatile, scalable, robust and open solutions to interfacing with living neurons

Motivated by the unexplored potential of in vitro neural systems for computing, and by the corresponding need of versatile, scalable interfaces for multimodal interaction, we present an accurate, modular, fully customizable and portable recording/stimulation solution that can be easily fabricated, robustly operated, and broadly disseminated. A reconfigurable platform that works across multiple industry standards enables a complete signal chain, from neural substrates sampled through high-density Micro-Electrode Arrays (MEAs) to data acquisition, downstream analysis and cloud storage. Built-in modularity supports the seamless integration of electrical/optical stimulation and fluidic interfaces. Custom MEA fabrication leverages maskless photolithography, favoring the rapid prototyping of a variety of configurations and spatial topologies. Through a native analysis and management software suite, the utility and robustness of our system is demonstrated across neural cultures and applications, including embryonic stem cell-derived and primary neurons, organotypic brain slices, 3D engineered tissue mimics, concurrent calcium imaging and long-term recording. Overall, our technology, termed Mind in Vitro to underscore the computing inspiration, provides an end-to-end solution that can be widely deployed due to its affordable (>10X cost-reduction) and open-source nature, catering to the expanding needs of both conventional and unconventional electrophysiology.

neuroscience↗

bioRxiv2023

On the mechanical origins of waving, coiling and skewing in Arabidopsis thaliana roots

By masterfully balancing directed growth and passive mechanics, plant roots are remarkably capable of navigating complex heterogeneous environments to find resources. Here we present a theoretical and numerical framework which allows us to interrogate and simulate the mechanical impact of solid interfaces on the growth pattern of plant organs. We focus on the well-known waving, coiling and skewing patterns exhibited by roots of Arabidopsis thaliana when grown on inclined surfaces, serving as a minimal model of the intricate interplay with solid substrates. By modelling growing slender organs as Cosserat rods that mechanically interact with the environment, our simulations verify hypotheses of waving and coiling arising from the combination of active gravitropism and passive root-plane responses. Skewing is instead related to intrinsic twist due to cell file rotation. Numerical investigations are outfitted with an analytical framework that consistently relates transitions between straight, waving, coiling and skewing patterns with substrate tilt angle. Simulations are found to corroborate theory and recapitulate a host of reported experimental observations, thus providing a systematic approach for studying in silico plant organs behavior in relation to their environment. SignificancePlant roots exhibit an exceptional ability to navigate in heterogeneous soil environments while overcoming obstacles. Our study combines theory and experimental observations to interrogate and simulate the mechanical impact of obstacles on organ growth. As a test case we focus on well-known observations of waving, coiling and skewing growth patterns of Arabidopsis thaliana roots grown on inclined substrates. Overall, our study explains a broad set of experimental observations through the minimal ingredients of gravitropism and passive mechanics. Our numerical framework provides an in silico laboratory, yielding quantitative insight into the dynamics of growing organs at the intersection of active processes and passive mechanics, applicable beyond plants to any slender growing system, from neurons or fungal hyphae to novel soft robots.

plant biology↗

bioRxiv2022

Microtubules self-repair in living cells

Microtubule self-repair has been studied both in vitro and in vivo as an underlying mechanism of microtubule stability. The turnover of tubulin dimers along the microtubule network has challenged the pre-existing dogma that only growing ends are dynamic. However, although there is clear evidence of tubulin incorporation into the shaft of polymerized microtubules in vitro, the possibility of such events taking place in living cells remains uncertain. In this study, we investigated this possibility by microinjecting purified tubulin dimers labeled with a red fluorophore into the cytoplasm of cells expressing GFP-tubulin. We observed the appearance of red dots along pre-existing green microtubule network within minutes. We found that the fluorescence intensities of these red dots were inversely correlated with the green signal, suggesting that the red dimers were incorporated into the microtubules and replaced the pre-existing green dimers. We then characterized the size and spatial frequency of these incorporations as a function of injected tubulin concentration and post-injection delay. The saturation of these measurements contradicted the hypothesis of nonspecific adsorption along microtubules and suggested that the injected dimers incorporated into a finite number of damaged sites. By our low estimate, within a few minutes of the injections, free dimers incorporated into major repair sites every 70 micrometers of microtubules. Finally, we mapped the location of these sites in micropatterned cells and found that they were more concentrated in regions where the actin filament network was less dense and where microtubules exhibited greater lateral fluctuations. These results provide evidences that microtubules do self-repair in living cells, and they provide a quantitative characterization of the temporal and spatial dynamics of this process in PtK2 cells.

cell biology↗

bioRxiv2020

Computationally assisted design and selection of maneuverable biological walking machines

The intriguing opportunities enabled by the use of living components in biological machines have spurred the development of a variety of muscle-powered bio-hybrid robots in recent years. Among them, several generations of bio-hybrid walkers have been established as reliable platforms to study untethered locomotion. However, despite these advances, such technology is not mature yet, and major challenges remain. This study takes steps to address two of them: the lack of systematic design approaches, common to bio-hybrid robotics in general, and in the case of bio-hybrid walkers specifically, the lack of maneuverability. We then present here a dual-ring biobot, computationally designed and selected to exhibit robust forward motion and rotational steering. This dual-ring biobot consists of two independent muscle actuators and a 4-legged scaffold asymmetric in the fore/aft direction. The integration of multiple muscles within its body architecture, combined with differential electrical stimulation, allows the robot to maneuver. The dual-ring robot design is then fabricated and experimentally tested, confirming computational predictions and turning abilities. Overall, our design approach based on modeling, simulation, and fabrication exemplified in this robot represents a route to efficiently engineer biological machines with adaptive functionalities.

bioengineering↗