Biology subjects

Colombo, L.

Publications and source records attributed to Colombo, L..

14 recordsLinked to original sources

bioRxiv2026

Newborns modulate and synchronise their Non-Nutritive Sucking behaviour to the temporal structure of sound

Sensorimotor synchronization, the ability to align motor behaviour with the temporal structure of external stimuli, is thought to emerge gradually during development, but its origins remain unclear. Here, we investigated whether newborns can adapt their sucking behaviour and synchronise it to the temporal structure of auditory stimuli within the first days of life. Forty-one healthy full-term newborns (24-72 hours old) were exposed to two three-minute auditory streams differing in temporal structure: a highly structured stream with a stable temporal pattern and a highly variant stream with greater temporal variability, while their non-nutritive sucking (NNS) was continuously recorded. We assessed changes in NNS frequency and pressure and quantified NNS-sound coupling using magnitude-squared coherence at frequencies distinctive to each auditory stream. Structured stimulation promoted a more organised NNS, with more suck bursts than the variant stream, and increased sucking vigour, with a transiently higher suck peak rate during the first minute relative to the variant stream and greater suck pressure relative to silence. Coherence analyses revealed that coherence was selectively higher at the frequencies distinctive to each auditory stream, both for the structured and the highly variant stimulus. These findings show that, within hours of birth, NNS behaviour can selectively align with stimulus-specific temporal regularities, even when these are weakly predictable and lack a dominant periodicity. This early capacity to extract temporal structure and coordinate motor behaviour may provide a foundation for the subsequent development of sensorimotor synchronisation.

neuroscience↗

bioRxiv2026

SEPALLATA MADS transcription factors act as key regulators in fertilization efficiency, ovule outer integument growth and mucilage secretory cell differentiation in Arabidopsis

In angiosperms, ovule development requires the activity of the C, D and E classes of MADS genes, which encode key transcriptional regulators of reproductive development. The SEPALLATA (SEP) MADS transcription factors (MTFs), which belong to the E class, act as organizing hubs of MADS heterotetrameric complexes and play an essential role in the development of flower organs. However, the role of the SEP genes in ovule and seed development has been difficult to determine due to redundancy in the subclade, the lack of observable phenotypes in single and double sep1 sep2 mutants and the homeotic conversion of the carpel into sepal or leaf in higher order sep mutants. Here, we engineered a version of SEP3 (SEP3{Delta}M) that encodes a protein lacking the DNA-binding MADS-domain but retains the oligomerization domains needed for MADS protein heterotetramerization. In vitro experiments demonstrated the ability of SEP3{Delta}M to interact with the C and D classes of MTF, reducing the capability of such MADS complex to efficiently bind DNA. sep3{Delta}M plants showed a delay in flower opening and organ maturation and a reduced fertility. The ovules exhibited reduced outer integument growth, and the few seeds that developed showed impaired mucilage secretion upon imbibition. RNA-seq analysis of sep3{Delta}M demonstrated misregulation of genes involved in outer integument and seed coat development. Taken together, these data indicate the key role of SEP3-containing MADS complexes in proper ovule outer integument growth and seed coat development.

plant biology↗

bioRxiv2026

Residual cortical responses and network reorganization in inherited retinal degeneration: electrophysiological evidence

Retinitis pigmentosa (RP) progressively deprives the retina of input, but whether the responsiveness of the visual cortex declines in parallel, remains preserved, or increases through compensatory gain remains unclear. Indeed, a weaker visually evoked response cannot, on its own, distinguish these possibilities, since it is equally compatible with a passively degraded input and with an actively recalibrated cortex. We combined spatially resolved steady-state visual evoked potentials (SSVEPs), which index stimulus-driven activity, with transcranial magnetic stimulation combined with electroencephalography (TMS-EEG), which probes cortical reactivity independently of vision, in patients with RP and in healthy controls. Nine patients (PTs) with RP (five women, age range 28 to 69 years) and nineteen sex-, age-, and handedness-matched healthy controls (thirteen women, mean age 42.6 years) were tested. They underwent SSVEP recordings to stimuli presented at three eccentricities (central, intermediate, peripheral) and single-pulse TMS-EEG over the left and right occipital cortex and, as a non-visual control site, the dominant motor cortex. We quantified SSVEP amplitude and phase at 12 Hz, early TMS-evoked potentials, oscillatory power, inter-trial phase synchrony, and functional connectivity and graph-theoretical network measures derived from the weighted phase lag index. SSVEP amplitude followed the expected central-to-peripheral gradient: PTs were comparable to healthy controls at the center, reduced but still above their own resting baseline at intermediate eccentricity, and no longer distinguishable from baseline in the periphery; phase differed from controls in a quarter of the central and half of the intermediate sectors. Occipital stimulation elicited a larger early negative deflection after left-hemisphere stimulation in PTs compared to controls, a stronger beta-band event-related spectral perturbation after stimulation of either hemisphere, and stronger, more efficiently distributed post-stimulus connectivity, despite comparable pre-stimulus connectivity, resting motor threshold, and most early evoked components. The pattern was site- and hemisphere-specific: left occipital stimulation produced widespread, mainly contralateral effects; right occipital stimulation a more circumscribed ipsilateral one, and motor cortex stimulation showed altered alpha-band activity without the bilateral occipital beta effect. Together, these results show that progressive retinal deafferentation in RP does not produce a parallel decline in cortical responsiveness. Visually driven activity weakens with eccentricity, while direct cortical perturbation reveals preserved and, at selected sites, enhanced reactivity. This dissociation is consistent with a homeostatic increase in cortical gain rather than a uniform loss of cortical function, and indicates that the deafferented cortex retains, and in places strengthens, its capacity to respond as retinal input deteriorates.

neuroscience↗

bioRxiv2025

ARABIDOPSIS Bsister and SEEDSTICK MADS-box transcription factors modulate maternal nutrient flow for seed development in Arabidopsis

Successful seed development in angiosperms depends on the coordinated transport and allocation of sugars from maternal tissues to the developing embryo and endosperm. In Arabidopsis thaliana, ovules function as carbohydrate sink organs, accumulating starch in both gametophytic and sporophytic domains prior to fertilization. This stored starch is later mobilized to support early embryogenesis. Despite extensive knowledge of starch metabolism in photosynthetic tissues, the regulatory mechanisms governing sugar transport in reproductive organs remain poorly understood. Recent studies have identified fertilization-dependent changes in nutrient flow, including callose-mediated modulation of symplastic transport at the phloem unloading site. However, the molecular players orchestrating these transitions are largely unknown. Here, we show that the MIKC MADS domain transcription factors ABS/TT16 and STK play critical roles in regulating maternal nutrient flow during ovule maturation and seed development. We dissect their functional redundancy using omics and genetic approaches, underscoring the importance of different ovule tissues in coordinating sugar transport pathways for post-fertilization development. Our findings reveal a previously underappreciated layer of genetic control over nutrient allocation in reproductive tissues and provide new insights into the metabolic reprogramming required for successful seed formation.

plant biology↗

bioRxiv2025

The transcription factor NO TRANSMITTING TRACT / WIP2 regulates cytokinin homeostasis in Arabidopsis

NO TRANSMITTING TRACT / WIP2 (NTT/WIP2) is a zinc finger transcription factor that alters developmental programs in Arabidopsis when overexpressed. Loss of NTT function, alone or in combination with closely related genes, leads to developmental defects in gynoecia and roots, respectively. Its overexpression phenotype resembles the effects of cytokinin treatments, suggesting a potential connection to this hormonal pathway. To further investigate this connection, we conducted cytokinin content and response evaluations, and transcriptome analyses in an inducible NTT line. Alterations in cytokinin content and response, as evaluated with the TCSn::GFP line, were observed. In both cases, as in the global expression analyses, the observed changes were dependent on the tissue analyzed. Moreover, Y1H and transactivation assays suggested direct NTT binding to the IPT5, AHP6 and CKX7, regulatory regions. We propose that NTT, among other mechanisms, regulates development through the modulation of cytokinin homeostasis at different levels, including its local concentration. Since NTT is a target of AUXIN RESPONSE FACTOR 5 / MONOPTEROS, the results presented here indicate that NTT connects an auxin input to a change in cytokinin levels.

plant biology↗

bioRxiv2025

SPOROCYTELESS/NOZZLE acts together with MADS-domain transcription factors to regulate an auxin-dependent network controlling the Megaspore Mother Cell development

The formation of the female gamete is a complex developmental process that begins with the differentiation of the Megaspore Mother Cell (MMC) within the ovule. SPOROCYTELESS/NOZZLE (SPL/NZZ) is the principal regulator of the MMC formation, as mutations in the SPL/NZZ gene lead to the failure of the MMC differentiation. Nonetheless, the SPL/NZZ-dependent regulatory pathway governing the MMC development remains largely unknown. Using a multi-omics approach, we identify direct SPL/NZZ targets and their downstream network. We discovered that SPL/NZZ interacts with ovule-identity MADS-domain transcription factor complexes, to regulate the expression of common target genes. By integrating the omics data with the analysis of either complementation or mutant lines, we describe a comprehensive regulatory mechanism, in which SPL/NZZ determines the differentiation of the MMC by acting on an auxin-dependent downstream network.

plant biology↗

bioRxiv2024

ESR2 orchestrates cytokinin dynamics leading to developmental reprogramming and green callus formation

Callus formation and shoot regeneration are natural plant abilities triggered by stress and damage. They are also key components of tissue culture, which for many species is crucial for gene editing, transformation, propagation, and other technologies, and their study provides valuable insights into plant development. The transcription factor ENHANCER OF SHOOT REGENERATION 2 (ESR2/DRNL/BOL/SOB) promotes green callus formation in roots and shoot regeneration when overactive, while the phythormone cytokinin plays a prominent role in both phenomena. Yet, the positive action of ESR2 on the cytokinin pathway had not been previously described. We explored the effects of ESR2 and found that cytokinin content and the expression of the cytokinin biosynthesis gene ISOPENTENYLTRANSFERASE 5 (IPT5) increase in plants where ESR2 activity is induced. ESR2 also regulates the cytokinin signaling repressor ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER 6 (AHP6), and surprisingly, the ESR2-stimulated green calli formation requires both IPT5 and AHP6. Therefore, ESR2 promotes both cytokinin biosynthesis and cytokinin signaling inhibition and requires a paradoxical combination of these two for green callus induction. This finding provides a foundation to better understand the processes involved in tissue reprogramming towards callus formation and the role of ESR2 in shoot regeneration and the development of new organs.

plant biology↗

bioRxiv2023

HISTONE DEACETYLASE 19 REGULATES SHOOT MERISTEMLESS EXPRESSION IN THE CARPEL MARGIN MERISTEM CONTRIBUTING TO OVULE NUMBER DETERMINATION AND TRANSMITTING TRACT DIFFERENTIATION

The gynoecium is critical for the reproduction of flowering species as it contains the ovules and the tissues required for pollen germination and guidance. These tissues are collectively known as the reproductive tract (ReT) and comprise stigma, style and transmitting tract (TT). The ovules and the ReT originate from a meristem within the pistil named carpel margin meristem (CMM). SHOOT MERISTEMLESS (STM) is a key transcription factor required for meristem formation and maintenance. In all above-ground meristems, including the CMM, STM has to be locally downregulated to allow proper organ differentiation. However, how this downregulation is achieved in the CMM is unknown. In this work, we have studied HISTONE DEACETYLASE 19 (HDA19) role in ovule and ReT differentiation, based on the observation that hda19-3 mutant displays reduced ovule number and fails to properly differentiate the TT. Fluorescence activated cell sorting (FACS) coupled with RNA-seq revealed that in the CMM of hda19-3 mutant, genes promoting organ development are downregulated while meristematic markers, including STM, are upregulated. We found that HDA19 is fundamental to downregulate STM in the CMM, allowing ovule formation and TT differentiation. STM is ectopically expressed in hda19-3 at intermediate stages of pistil development, and its downregulation by RNA interference alleviated hda19-3 phenotypic defects. Furthermore, chromatin immunoprecipitation assays indicated that STM is a direct target of HDA19 during pistil development and that SEEDSTICK (STK) is required for the histone acetylation-mediated regulation of STM. Our results have led to the identification of the factors required for STM silencing in the gynoecium allowing organogenesis and tissue differentiation from the CMM.

plant biology↗

bioRxiv2022

MADS31 supports female germline development by repressing the post-fertilization program in cereal ovules

The female germline of flowering plants develops within a niche of somatic ovule cells, also referred to as the nucellus. How niche cells maintain their own somatic developmental program, yet support the development of adjoining germline cells, remains largely unknown. Here we report that MADS31, a conserved MADS-box transcription factor from the B-sister subclass, is a potent regulator of niche cell identity in barley. MADS31 is preferentially expressed in nucellar cells directly adjoining the germline, and loss-of-function mads31 mutants exhibit deformed and disorganized nucellar cells, leading to impaired germline development and partial female sterility. Molecular assays indicate that MADS31 encodes a potent transcriptional repressor, repressing genes in the ovule that are normally active in the seed. One prominent target of MADS31 is NRPD4b, a seed-expressed component of RNA polymerase IV/V that is involved in gene silencing via RNA directed DNA methylation. NRPD4b is directly repressed by MADS31 in vivo and is de-repressed in mads31 ovules, while overexpression of NRPD4b recapitulates the mads31 ovule phenotype. This coincides with specific changes in histone methylation and is consistent with NRPD4b being directly repressed by MADS31 to maintain ovule niche functionality. Our findings reveal a new mechanism by which somatic ovule tissues maintain their own identity before transitioning to the post-fertilization program.

plant biology↗

bioRxiv2022

Cholesterol-laden brain-permeable nanoparticles support long-lasting cognitive recovery and motor amelioration in the slow-progressing zQ175DN mouse model of Huntington's Disease

Evidence that Huntingtons disease (HD) is characterized by impaired cholesterol biosynthesis in the brain has led to strategies to increase its level in the brain of the rapidly progressing R6/2 mouse model, with a positive therapeutic outcome. Here we tested the long-term efficacy of chronic administration of cholesterol to the brain of the slowly progressing zQ175DN knock-in HD mice in preventing ("early treatment") or reversing ("late treatment") HD symptoms. To do this we used the most advanced formulation of cholesterol loaded brain-permeable nanoparticles (NPs), termed hybrid-g7-NPs-chol, which were injected intraperitoneally. We show that one cycle of treatment with hybrid-g7-NPs-chol, administered in the presymptomatic ("early treatment") or symptomatic ("late treatment") stages is sufficient to normalize cognitive defects up to 5 months, as well as to improve other behavioral and neuropathological parameters. A multiple cycle treatment combining both early and late treatments ("2 cycle treatment") lasting 6 months generates therapeutic effects for more than 11 months, without severe adverse reactions. Sustained cholesterol delivery to the brain of zQ175DN mice also reduces mutant Huntingtin aggregates in both the striatum and cortex and completely normalizes synaptic communication in the striatal medium spiny neurons compared to saline-treated HD mice. Furthermore, through a meta-analysis of published and current data, we demonstrated the power of hybrid-g7-NPs-chol and other strategies able to increase brain cholesterol biosynthesis, to reverse cognitive decline and counteract the formation of mutant Huntingtin aggregates. These results demonstrate that cholesterol delivery via brain-permeable NPs is a therapeutic option to sustainably reverse HD-related behavioral decline and neuropathological signs over time, highlighting the therapeutic potential of cholesterol-based strategies in HD patients.

neuroscience↗

bioRxiv2021

Mutant huntingtin crosses the neuromuscular junction and causes transmission-selective pathological alterations in huntingtin expressing myotubes

A potential explanation for the spatiotemporal accumulation of pathological lesions in the brain of patients with neurodegenerative protein misfolding diseases (PMDs) is cell-to-cell transmission of aggregation-prone, misfolded proteins. Little is known about central to peripheral transmission and its contribution to pathology. We show that transmission of Huntingtons disease- (HD-) associated mutant HTT exon 1 (mHTTEx1) occurs across the neuromuscular junctions in human iPSC cultures and in vivo in wild-type mice. We found that transmission is an active and dynamic process, that happens prior to aggregate formation and is regulated by synaptic activity. Furthermore, we find that transmitted mHTTEx1 causes HD-relevant pathology at a molecular and functional level in human muscle cells, even in the presence of ubiquitous expression mHTTEx1. With this work we uncover a casual-link between mHTTEx1 synaptic transmission and pathology, highlighting the therapeutic potential in blocking toxic protein transmission in PMDs.

neuroscience↗

bioRxiv2020

Insights into kinetics, release, and behavioral effects of brain-targeted hybrid nanoparticles forcholesterol delivery in Huntington disease

Supplementing brain cholesterol is emerging as a potential treatment for Huntingtons disease (HD), a genetic neurodegenerative disorder characterized, among other abnormalities, by inefficient brain cholesterol biosynthesis. However, delivering cholesterol to the brain is challenging due to the bloodbrain barrier (BBB), which prevents it from reaching the striatum, especially, with therapeutically relevant doses. Here we describe the distribution, kinetics, release, and safety of novel hybrid polymeric nanoparticles made of PLGA and cholesterol which were modified with an heptapeptide (g7) for BBB transit (hybrid-g7-NPs-chol). We show that these NPs rapidly reach the brain and target neural cells. Moreover, deuterium-labeled cholesterol from hybrid-g7-NPs-chol is released in a controlled manner within the brain and accumulates over time, while being rapidly removed from peripheral tissues and plasma. We confirm that systemic and repeated injections of the new hybrid-g7-NPs-chol enhanced endogenous cholesterol biosynthesis, prevented cognitive decline, and ameliorated motor defects in HD animals, without any inflammatory reaction. In summary, this study provides insights about the benefits and safety of cholesterol delivery through advanced brain-permeable nanoparticles for HD treatment.

neuroscience↗

bioRxiv2020

Role of misfolded tau in the onset and progression of brain toxicity after trauma

Traumatic brain injury (TBI) is associated with widespread tau pathology in about one third of patients. We previously found that TBI induces a transmissible tau pathology (tauTBI), with late cognitive decline and synaptic dysfunction. To understand whether tauTBI is a marker of ongoing neurodegeneration or a driver of functional decline, we employed C. elegans. Brain homogenates from chronic TBI mice, or from mice in which tauTBI had been transmitted by intracerebral inoculation, impaired C. elegans motility and neuromuscular synaptic transmission. Brain homogenates from tau P301L transgenic mice, or pre-aggregated recombinant tau, induced a similar toxic response. Protease digestion or pre-incubation of homogenates with anti-tau antibodies abolished toxicity, and TBI brain homogenates from tau knock-out mice had no toxic effect. These results support a vital role of abnormal tau species in chronic neurodegeneration after TBI and set the groundwork for the development of a C. elegans-based platform for screening anti-tau compounds.

neuroscience↗

bioRxiv2019

A peptide pair coordinates regular ovule initiation patterns with seed number and fruit size

Ovule development in Arabidopsis thaliana involves pattern formation which ensures that ovules are regularly arranged in the pistils to reduce competition for nutrients and space. Mechanisms underlying pattern formation in plants, such as phyllotaxis, flower morphogenesis or lateral root initiation, have been extensively studied, and genes controlling the initiation of ovules have been identified. However, how a regular spacing of ovules is achieved is not known. Using natural variation analysis combined with quantitative trait locus analysis, we found that the spacing of ovules in the developing fruits is controlled by two secreted peptides, EPFL2 and EPFL9 (also known as Stomagen), and their receptors from the ERECTA (ER) family that act from the carpel wall and the placental tissue. We found that a signalling pathway controlled by EPFL9 acting from the carpel wall through the LRR-receptor kinases ER, ERL1 and ERL2 promotes fruit growth. Regular spacing of ovules depends on EPFL2 expression in the carpel wall and in the inter-ovule spaces, where it acts through ERL1 and ERL2. Loss of EPFL2 signalling results in shorter fruits and irregular spacing of ovules or even ovule twinning. The EPFL2 expression pattern between ovules is under negative-feedback regulation by auxin, which accumulates in the arising ovule primordia. We propose that the auxin-EPFL2 signalling module evolved to control the initiation and regular, equidistant spacing of ovule primordia, which serves to minimise competition between developing seeds. Together, EPFL2 and EPFL9 coordinate ovule patterning and thereby seed number with fruit growth through a set of shared receptors.

plant biology↗