Biology subjects

Montanari, R.

Publications and source records attributed to Montanari, R..

7 recordsLinked to original sources

bioRxiv2026

A mobile optical coherence microscope for studying aquatic organisms in- and outside the laboratory

High-resolution, three-dimensional imaging of live organisms has largely depended on laboratory-bound microscopes, limiting quantitative analysis of morphology and dynamics to species that survive transport and thrive under lab-controlled conditions. Here we present a mobile optical coherence microscopy (OCM) platform that overcomes this constraint, delivering [~]2.5 {micro}m axial resolution, label-free, volumetric imaging of live aquatic organisms in both laboratory and remote field environments. We demonstrate the platform across a broad range of aquatic organisms from the lab and field, spanning multiple phyla - including cnidarians, poriferans, annelids, arthropods and echinoderms - resolving internal anatomy, tissue boundaries and organismal morphology at micrometer scale without fixation, fluorescent labeling, or specialized sample preparation. High-speed acquisition, with up to 250 kHz A-line rate and 7.7 Hz volume rate, further enabled morphodynamic imaging of live biological processes, including cellular aggregate motility, embryonic cell division, and organ-level peristaltic dynamics in intact, living animals. To demonstrate field deployability, the platform was operated during the EMBL TREC pan-European expedition, enabling on-site, label-free imaging of marine organisms and plankton immediately upon collection. By decoupling high-resolution volumetric imaging from fixed laboratory settings, mobile OCM opens a path toward field-ready quantitative morphological and dynamic phenotyping of aquatic life.

bioengineering↗

bioRxiv2025

Pupillometry Reveals Autonomic Adjustments During Diving Reflex in Face Immersion Apnea

The human diving reflex is an innate cardiorespiratory adjustment triggered during apnea that concurrently activates both sympathetic and parasympathetic branches of the autonomic nervous system to maintain physiological stability under stress. Although pupil dilation and constriction are antagonistically regulated by these branches, the effect of the diving reflex on pupil diameter oscillations, known as hippus, remains unexplored. Here, we compared hippus in healthy participants during breathing or apnea either with (Wet) or without (Dry) facial immersion in cold water. In both apneic conditions, hippus exhibited reduced power in the low-frequency band (< 0.25 Hz). Notably, during Wet apnea, we observed a reallocation of power towards higher frequencies (> 0.25 Hz) and increased entropy of fluctuations, indicating a shift in autonomic balance and greater signal complexity during the diving reflex. This pilot study highlights pupil dynamics as a sensitive and non-invasive probe of autonomic adjustments underlying the human diving reflex.

physiology↗

bioRxiv2022

Callosal inputs generate side-invariant receptive fields in the barrel cortex

Barrel cortex integrates contra- and ipsilateral whiskers inputs. While contralateral inputs depend on the thalamocortical innervation, ipsilateral ones are thought to rely on callosal axons. These are more abundant in the barrel cortex region bordering with S2 and containing the row A-whiskers representation, the row lying nearest to the facial midline. Here we ask what role this callosal axonal arrangement plays in ipsilateral tactile signalling. We found that novel object exploration with ipsilateral whiskers confines c-Fos expression within the highly callosal subregion. Targeting this area with in vivo patch-clamp recordings revealed neurons with uniquely strong ipsilateral responses dependent on the corpus callosum, as assessed by tetrodotoxin silencing and by optogenetic activation of the contralateral hemisphere. Still, in this area, stimulation of contra- or ipsilateral row A-whiskers evoked an indistinguishable response in some neurons, mostly located in layers 5/6, indicating their involvement in the midline representation of the whiskers sensory space.

neuroscience↗

bioRxiv2021

A Focused Small-Molecule Screen Identifies PHA-680626 as an Amphosteric Inhibitor Disrupting the Interaction between Aurora-A and N-Myc

Neuroblastoma is a severe childhood disease, accounting for ~10% of all infant cancers. The amplification of the MYCN gene, coding for the N-Myc transcriptional factor, is an essential marker correlated with tumor progression and poor prognosis. In neuroblastoma cells, the mitotic kinase Aurora-A (AURKA), also frequently overexpressed in cancer, prevents N-Myc degradation, by directly binding to a highly conserved N-Myc region, i.e. Myc Box I. As a result, elevated levels of N-Myc, which are required for the growth of MYCN amplified cells, are observed. During the last years, it has been demonstrated that the ATP competitive inhibitors of AURKA CD532, MLN8054 and Alisertib also cause essential conformational changes in the structure of the activation loop of the kinase that prevent N-Myc binding, thus impairing the formation of the AURKA/N-Myc complex. In this study, starting from a screening of crystal structures of AURKA in complex with known inhibitors, we identified additional compounds affecting the conformation of the kinase activation loop. We assessed the ability of such compounds to disrupt the interaction between AURKA and N-Myc in vitro, using Surface Plasmon Resonance competition assays, and in tumor cell lines overexpressing MYCN, by performing Proximity Ligation Assays. Finally, their effects on N-Myc cellular levels and cell viability were investigated. Our results, identifying PHA-680626 as an amphosteric inhibitor both in vitro and MYCN overexpressing cell lines, expand the repertoire of known conformational disrupting inhibitors of the AURKA/N-Myc complex, and confirm that altering the conformation of the activation loop of AURKA with a small molecule is an effective strategy to destabilize the AURKA/N-Myc interaction in neuroblastoma cancer cells.

cancer biology↗

bioRxiv2020

Cytosolic localization and in vitro assembly of human de novo thymidylate synthesis complex

De novo thymidylate synthesis is a crucial pathway for normal and cancer cells. Deoxythymidine monophosphate (dTMP) is synthesized by the combined action of three enzymes: serine hydroxymethyltransferase (SHMT), dihydrofolate reductase (DHFR) and thymidylate synthase (TYMS), the latter two targets of widely used chemotherapeutics such as antifolates and 5-fluorouracil. These proteins translocate to the nucleus after SUMOylation and are suggested to assemble in this compartment into the thymidylate synthesis complex (dTMP-SC). We report the intracellular dynamics of the complex in lung cancer cells by in situ proximity ligation assay, showing that it is also detected in the cytoplasm. This result strongly indicates that the role of the dTMP-SC assembly may go beyond dTMP synthesis. We have successfully assembled the dTMP synthesis complex in vitro, employing tetrameric SHMT1 and a bifunctional chimeric enzyme comprising human TYMS and DHFR. We show that the SHMT1 tetrameric state is required for efficient complex assembly, indicating that this aggregation state is evolutionary selected in eukaryotes to optimize protein-protein interactions. Lastly, our results on the activity of the complete thymidylate cycle in vitro, may provide a useful tool to develop drugs targeting the entire complex instead of the individual components.

biochemistry↗

bioRxiv2020

Slow Wave Oscillation reveals the discrete segregation of dorsal striatum

Behavioural studies differentiate the rodent dorsal striatum (DS) into lateral and medial regions based on their particular roles; however, anatomical evidence suggest that it is a unified structure. To understand striatal dynamics and basal ganglia functions, it is essential to clarify the circuitry that supports this behavioural-based segregation. Here, we show that the mouse DS is made of two non-overlapping functional circuits divided by a sharp boundary. Combining in vivo optopatch-clamp and extracellular recordings of spontaneous activity, we demonstrate different coupling of lateral and medial striatum to the cortex together with an independent integration of the slow and fast oscillations, due to particular cortico-striatal connectivity and local properties of each region. Additionally, distinct properties between striatonigral and striatopallidal neurons were revealed along DS. These results demonstrate that the rodent DS is segregated in two neuronal circuits, in homology with the caudate and putamen nuclei of primates.

neuroscience↗

bioRxiv2018

Visual salience of the stop-signal affects neural dynamics of controlled inhibition

The countermanding or stop-signal task is broadly used to evaluate response inhibition: it sporadically requires to inhibit a movement upon an incoming salient stop-signal.\n\nTo study the neural basis of arm movements inhibition we combined the approach typically employed for the study of perceptual-decision making with the countermanding task, that is broadly used to evaluate response inhibition\n\nTo this aim we modified the salience of the stop-signal and we found that this modulation affected the ability to inhibit in macaque monkeys: coherently to what already observed in humans, we found that less salient stimuli deteriorate inhibitory performance. These behavioural results were subtended by neural modulations representing an inhibitory process that started later in time and showed a less steeper dynamic for stimuli difficult to be processed.\n\nThis study shows that the neural patterns observed when deciding to stop are broadly similar to the neural patterns observed when deciding to act in the literature; thus it is a first step in investigating the perceptual decision making process involved in movement inhibition.

neuroscience↗