Biology subjects

Leite, M.

Publications and source records attributed to Leite, M..

14 recordsLinked to original sources

bioRxiv2026

Widespread occurrence of XXY females in Drosophila mojavensis and related species

Bridges first reported that XXY individuals are fertile females in Drosophila melanogaster, arising at low frequency (~1/2000) through errors in chromosome segregation (BRIDGES 1914; BRIDGES 1916). Although widely used in genetic studies, XXY females are seldom found in wild-type strains or natural populations, presumably because natural selection keeps them at very low frequency. Here we show that many wild-type strains of D. mojavensis and its close relatives D. arizonae, D. navojoa, and D. wheeleri carry XXY and XXYY females at frequencies of up to 88%; males from these strains are frequently XYY or XYYY. Although field collections were hampered by population declines and safety concerns, we found one XXY female (out of 87) and one XYY male (out of 36) in a natural population of D. mojavensis, indicating that these genotypes also occur at appreciable frequencies in nature. XXY females are expected to produce XY-bearing and X-bearing eggs in equal proportions; instead, we found a 57:43 ratio. Mathematical modeling shows that this meiotic-drive advantage is sufficient to explain high frequencies of XXY females. It is universally accepted that Y evolution responds exclusively to male selection, yet in XXY-bearing species it also responds to female fitness.

evolutionary biology↗

bioRxiv2026

The anxiolytic-like effect of the aqueous extract of Punica granatum depends on the stress level of fish and mice: Evidence of a homeostatic re-establishing effect

Pomegranate (Punica granatum) has shown potential anxiolytic and antidepressant effects in preclinical research, but its mechanisms remain unclear. Importantly, some evidence suggests that the effects of P. granatum depend on previous stress, suggesting an adaptogenic effect. This study aimed to evaluate the anxiolytic-like effects of an aqueous extract of P. granatum (AE-PG) on lambaris and zebrafish exposed to acute restraint stress (ARS) and female BALB/c mice exposed to unpredictable chronic mild stress (CUMS). In addition, we evaluated antidepressant-like effects in female BALB/c mice. Acute administration of AE-PG (1 mg/kg, i.p.) was tested in the novel tank test (NTT) in lambaris and zebrafish and in the light/dark test (LDT) in zebrafish, while chronic (7 days) administration of AE-PG (1 mg/kg, i.p.) was tested in the elevated plus-maze (EPM), open-field test (OFT), splash test (ST), and forced swim test (FST) in mice. ARS increased geotaxis and erratic swimming in the NTT, as well as scototaxis, risk assessment, and erratic swimming in the LDT. Acute pre-treatment with AE-PG blocked the effects of ARS on these anxiety-like endpoints in both lambaris and zebrafish. CUMS increased open arm avoidance in the EPM, thigmotaxis and horizontal and vertical exploration in the OFT, and immobility in the FST, and decreased grooming in the ST. Chronic treatment with AE-PG blocked the effects of CUMS on anxiety-like endpoints, and depression-like endpoints, in mice. An in silico analysis of the main compounds in AE-PG suggested a possible mechanism of action on a wide network of targets that converge on PI3K/Akt and show differential expression in brain regions associated with fear- and anxiety-like behavior. Molecular docking suggests that a compound from AE-PG, ellagic acid, is likely to act through estrogen receptors, which could help reestablish homeostatic balance in response to stress.

pharmacology and toxicology↗

bioRxiv2025

NDNF+ interneurons have a privileged role in regulating cortical excitability

Cortical processing depends critically on the precise modulation of excitatory activity by a diverse population of GABAergic interneurons. Among these, neuron-derived neurotrophic factor-expressing (NDNF+) interneurons have been proposed to be "master regulators" of cortical microcircuits. While their role in coordinating physiological cortical activity is beginning to be understood, their contribution to restraining hyperexcitability during epileptiform activity remains unexplored. To address this, we employed calcium imaging, optogenetics, and chemogenetics in an NDNF-Cre mouse line to investigate how NDNF+ interneurons influence cortical hyperactivity and spontaneous seizures. Our results demonstrate that NDNF+ interneurons are actively recruited during interictal spikes and focal seizures, although more slowly than parvalbumin-positive interneurons. Optogenetic hyperpolarization of NDNF+ interneurons exacerbates epileptic discharges, whereas their depolarization suppresses focal seizures, even when the optogenetic stimulus is delayed by several seconds from seizure onset. This effect is largely mediated by GABAB receptor signalling. Additionally, chemogenetic depolarization of NDNF+ interneurons has robust antiepileptic effects both ex vivo and in vivo. Collectively, these findings establish NDNF+ neurons as key regulators of cortical excitability and identify them as promising cellular targets for the development of anti-epileptic therapies.

neuroscience↗

bioRxiv2025

Bridging Light and Sound: a Spironaphtopyran-Rhodamine Dyad with High-Contrast Photoswitching Between Fluorescence and Photoacoustic Signal.

Fluorescence and photoacoustic imaging are complementary modalities that provide distinct advantages for biological imaging: fluorescence microscopy offers high sensitivity and resolution, while photoacoustic imaging enables deeper penetration in complex tissue. Leveraging the strengths of both modalities through optically switchable contrast agents can offer enhanced imaging contrast and facilitate dual-modality imaging. Here, we report a photoswitchable probe capable of toggling between high fluorescence and high photoacoustic signal upon illumination, exploiting Forster Resonance Energy Transfer (FRET). We engineer novel spironaphtopyran photoswitches which undergo reversible photoisomerization between absorbing and non-absorbing states. Their photoswitching properties were systematically characterized, establishing structure-properties relationships, and providing the first photoacoustic investigation into this class of compounds. The best-performing switch was incorporated into a FRET dyad with a rhodamine fluorophore, which exhibits robust, reversible switching between fluorescent and photoacoustic-dominant states with excellent contrast in vitro, establishing a foundation for multimodal imaging probes with promising potential for dynamic correlative imaging.

biochemistry↗

bioRxiv2023

Anti-seizure Gene Therapy for Focal Cortical Dysplasia

Focal Cortical Dysplasias (FCDs) are a common subtype of malformation of cortical development, which frequently present with a spectrum of cognitive and behavioural abnormalities as well as pharmacoresistant epilepsy. FCD type II is typically caused by somatic mutations resulting in mTOR hyperactivity, and is the commonest pathology found in children undergoing epilepsy surgery. However, surgical resection does not always result in seizure freedom, and is often precluded by proximity to eloquent brain regions. Gene therapy is a promising potential alternative treatment and may be appropriate in cases that represent an unacceptable surgical risk. Here, we evaluated a gene therapy based on overexpression of the Kv1.1 potassium channel in a mouse model of frontal lobe FCD. An engineered potassium channel (EKC) transgene was placed under control of a human promoter that biases expression towards principal neurons (CAMK2A) and packaged in an adeno-associated viral vector (AAV9). We used an established FCD model generated by in utero electroporation of frontal lobe neural progenitors with a constitutively active human RHEB plasmid, an activator of mTOR Complex 1. First, we further characterised this by quantifying electrocorticograms and behavioural abnormalities, both in mice developing spontaneous generalised seizures and in mice only exhibiting abnormal interictal discharges. Then, using continuous video-electrocorticogram recordings from epileptic mice before and after injection of AAV9-CAMK2A-EKC in the dysplastic region, we observed a robust decrease in the frequency of seizures and in interictal activity, compared to mice injected with a control viral vector. Despite the robust anti-epileptic effect of the treatment, there was neither an improvement nor a worsening of performance in behavioural tests sensitive to frontal lobe function. AAV9-CAMK2A-EKC had no effect on interictal activity or behaviour in non-epileptic mice. AAV9-CAMK2A-EKC gene therapy is a promising therapy with translational potential to treat the epileptic phenotype of mTOR-related malformations of cortical development. Cognitive and behavioural co-morbidities may, however, resist an intervention aimed at reducing circuit excitability.

neuroscience↗

bioRxiv2021

Alpha-synuclein is present in the nucleus in human brain tissue and is pathologically modified in Dementia with Lewy Bodies

Dementia with Lewy bodies is pathologically defined by the cytoplasmic accumulation of alpha-synuclein within neuronal cells in the brain. Alpha-synuclein is predominately pre-synaptic, but has been reported present in various subcellular compartments in cell and animal models. In particular, nuclear alpha-synuclein is evident in-vitro and in disease models and has been associated with altered DNA integrity, gene transcription, nuclear homeostasis. However, owing to various factors, the presence of alpha-synuclein in the nuclei of human brain cells remains controversial, as does its role in synucleinopathies. Here, we close this gap and provide a unique demonstration confirming the presence of nuclear alpha-synuclein in post-mortem brain tissue obtained from cases of dementia with Lewy bodies as well as from controls via immunohistochemistry, immunoblot, and label-free mass-spectrometry. Discrete intra-nuclear alpha-synuclein puncta reactive against phosphorylated serine 129-alpha-synuclein and pan-alpha-synuclein antibodies were observed in cortical neurons and non-neuronal cells in fixed brain sections and in isolated nuclear preparations from Dementia with Lewy bodies cases and matched controls. Subsequent biochemical analysis of subcellular fractionated tissue confirmed alpha-synuclein as present in a nuclear fraction at levels ~ 10-fold lower than in the cytoplasm. Critically, however, an increase in monomeric nuclear alpha-synuclein phosphorylated as serine 129 was observed in cases of dementia with Lewy bodies alongside higher molecular weight pan- and phosphorylation reactive alpha-synuclein species, consistent with the formation of intranuclear phosphorylated alpha-synuclein oligomers. Furthermore, the presence of nuclear alpha-synuclein was confirmed via label free mass spectrometry, as 6 unique alpha-synuclein derived peptide sequences were identified in nuclear fractions (71.4% sequence coverage). Collectively, our data confirm the presence of nuclear alpha-synuclein in human brain tissue and describe nuclear pathology associated with dementia with Lewy bodies. These findings address a major controversy in the synucleinopathy field by confirming the presence of nuclear alpha-synuclein in autoptic human brain tissue and, for the first time, identify that alpha-synuclein is aggregated into novel and potentially pathological assemblies in the nucleus as part of the disease process associated with dementia with Lewy bodies and thus may contribute to the disease phenotype.

pathology↗

bioRxiv2020

Extracellular glutamate and GABA transients at the transition from interictal spiking to seizures

Focal epilepsy is associated with intermittent brief population discharges (interictal spikes), which resemble sentinel spikes that often occur at the onset of seizures. Why interictal spikes self-terminate whilst seizures persist and propagate is incompletely understood. We used fluorescent glutamate and GABA sensors in an awake rodent model of neocortical seizures to resolve the spatiotemporal evolution of both neurotransmitters in the extracellular space. Interictal spikes were accompanied by brief glutamate transients which were maximal at the initiation site and rapidly propagated centrifugally. GABA transients lasted longer than glutamate transients and were maximal [~]1.5 mm from the focus where they propagated centripetally. Prior to seizure initiation GABA transients were attenuated, whilst glutamate transients increased, consistent with a progressive failure of local inhibitory restraint. As seizures increased in frequency, there was a gradual increase in the spatial extent of spike-associated glutamate transients associated with interictal spikes. Neurotransmitter imaging thus reveals a progressive collapse of an annulus of feed-forward GABA release, allowing seizures to escape from local inhibitory restraint.

neuroscience↗

bioRxiv2020

Learning to live with Dale's principle: ANNs with separate excitatory and inhibitory units

AO_SCPLOWBSTRACTC_SCPLOWThe units in artificial neural networks (ANNs) can be thought of as abstractions of biological neurons, and ANNs are increasingly used in neuroscience research. However, there are many important differences between ANN units and real neurons. One of the most notable is the absence of Dales principle, which ensures that biological neurons are either exclusively excitatory or inhibitory. Dales principle is typically left out of ANNs because its inclusion impairs learning. This is problematic, because one of the great advantages of ANNs for neuroscience research is their ability to learn complicated, realistic tasks. Here, by taking inspiration from feedforward inhibitory interneurons in the brain we show that we can develop ANNs with separate populations of excitatory and inhibitory units that learn just as well as standard ANNs. We call these networks Dales ANNs (DANNs). We present two insights that enable DANNs to learn well: (1) DANNs are related to normalization schemes, and can be initialized such that the inhibition centres and standardizes the excitatory activity, (2) updates to inhibitory neuron parameters should be scaled using corrections based on the Fisher Information matrix. These results demonstrate how ANNs that respect Dales principle can be built without sacrificing learning performance, which is important for future work using ANNs as models of the brain. The results may also have interesting implications for how inhibitory plasticity in the real brain operates.

neuroscience↗

bioRxiv2020

EMBRYONIC DEVELOPMENT OF THE FIRE-EYE-TETRA Moenkhausia oligolepis (CHARACIFORMES: CHARACIDAE)

This study describes the embryonic development of Moenkhausia oligolepis in captive conditions. After fertilization, the embryos were collected every 10 min up to 2 h, every 20 min up to 4 h, and every 30 min until hatching. The fertilized eggs of M. oligolepis measured approximately 0.85 {+/-} 0.5 mm and have an adhesive surface. The embryonic development lasted 14 hours at 25{degrees}C, with the Zygote, Cleavage, Blastula, Gastrula, Neurula and Segmentation phases. The hatching occurred in embryos around the 30-somites stage. Our results bring only the second description of embryonic development to a species of Moenkhausia genus, the first for the refereed species. Such data are of paramount importance considering the current conflicting state of this genus phylogenetic classification and may help taxonomic studies. Understand the biology of a species that is easily handling in captive conditions and has an ornamental appeal may assist studies in its reproduction in order to both, supply the aquarium market and help the species conservation in nature. Moreover, our data enable the M. oligolepis to be used as a model species in biotechnological applications, such germ cell transplantation approach.

developmental biology↗

bioRxiv2019

LGI1 downregulation increases neuronal circuit excitability

LGI1 (Leucine-Rich Glioma-Inactivated 1) is a secreted trans-synaptic protein that interacts presynaptically with Kv1.1 potassium channels and ADAM23, and postsynaptically, influencing AMPA receptors through a direct link with the ADAM22 cell adhesion protein. Haploinsufficiency of LGI1 or autoantibodies directed against LGI1 are associated with human epilepsy, generating the hypothesis that a subacute reduction of LGI1 is sufficient to increase network excitability. We tested this hypothesis in ex vivo hippocampal slices and in neuronal cultures, by subacutely reducing LGI1 expression with shRNA. Injection of shRNA-LGI1 in the hippocampus increased dentate granule cell excitability and low frequency facilitation of mossy fibers to CA3 pyramidal cell neurotransmission. Application of the Kv1 family blocker, alpha-dendrotoxin, occluded this effect, implicating the involvement of Kv1.1. This subacute reduction of LGI1 was also sufficient to increase neuronal network activity in neuronal primary culture. These results indicate that a subacute reduction in LGI1 potentiates neuronal excitability and short-term synaptic plasticity, and increases neuronal network excitability, opening new avenues for the treatment of limbic encephalitis and temporal lobe epilepsies. Significant StatementDown-regulation of LGI1 protein increases excitability at mossy fiber to CA3 pyramidal cell synapses and in hippocampal primary cultures. These alterations alter network function and so could contribute to symptoms observed in patients affected by limbic encephalitis.

neuroscience↗

bioRxiv2018

A genetically encoded fluorescent sensor for in vivo imaging of GABA

Current techniques for monitoring GABA, the primary inhibitory neurotransmitter in vertebrates, cannot follow ephemeral transients in intact neural circuits. We applied the design principles used to create iGluSnFR, a fluorescent reporter of synaptic glutamate, to develop a GABA sensor using a protein derived from a previously unsequenced Pseudomonas fluorescens strain. Structure-guided mutagenesis and library screening led to a usable iGABASnFR ({Delta}F/Fmax ~ 2.5, Kd ~ 9 M, good specificity, adequate kinetics). iGABASnFR is genetically encoded, detects single action potential-evoked GABA release events in culture, and produces readily detectable fluorescence increases in vivo in mice and zebrafish. iGABASnFR enabled tracking of: (1) mitochondrial GABA content and its modulation by an anticonvulsant; (2) swimming-evoked GABAergic transmission in zebrafish cerebellum; (3) GABA release events during inter-ictal spikes and seizures in awake mice; and (4) GABAergic tone decreases during isoflurane anesthesia. iGABASnFR will permit high spatiotemporal resolution of GABA signaling in intact preparations.

neuroscience↗

bioRxiv2018

Semiology, clustering, periodicity and natural history of seizures in an experimental visual cortical epilepsy model

ObjectiveTo characterize a rat model of focal neocortical epilepsy for use in developing novel therapeutic strategies in a type of epilepsy that represents a significant unmet need.\n\nMethodsIntracortical tetanus toxin (TeNT) injection was used to induce epilepsy in rats. Seizures and their behavioural manifestations were evaluated with continuous video-electrocorticography telemetry.\n\nResultsTeNT injection into rat primary visual cortex induced focal neocortical epilepsy without preceding status epilepticus. The latency to first seizure ranged from 3 to 7 days. Seizure duration was bimodal, with both short (approximately 30s) and long-lasting (>100s) seizures occurring in the same animals. Seizures were accompanied by non-motor features such as behavioural arrest, or motor seizures with or without evolution to generalized tonic-clonic seizures. Seizures were commoner during the sleep phase of a light-dark cycle. Seizure occurrence was not random, and tended to cluster with significantly higher probability of recurrence within 24 hours of a previous seizure. Across animals, the number of seizures in the first week could be used to predict the number of seizures in the following 22 days.\n\nSignificanceThe TeNT model of visual cortical epilepsy is a robust model of acquired focal neocortical epilepsy, and is well suited for preclinical evaluation of novel anti-epileptic strategies. We provide here a detailed analysis of the epilepsy phenotype, seizure activity, electrographic features, and the semiology. In addition we provide a predictive framework that can be used to reduce variation and consequently animal use in pre-clinical studies of potential treatments.\n\nKey PointsO_LITetanus toxin injection into rat visual cortex induces focal cortical epilepsy.\nC_LIO_LIElectrographic seizures were associated with non-motor and motor features with or without evolution to generalized tonic-clonic seizures.\nC_LIO_LISeizures could not be provoked by intermittent photic stimulation.\nC_LIO_LISeizures were clustered in time and exhibited a circadian variation in frequency.\nC_LIO_LIThe number of seizures in first week after seizure onset could be used to predict the total number of seizures in the following 3 weeks.\nC_LI

neuroscience↗

bioRxiv2018

Dendritic NMDA receptors in parvalbumin neurons enable strong and stable neuronal assemblies

Parvalbumin-expressing (PV+) GABAergic interneurons mediate feedforward and feedback inhibition and have a key role in gamma oscillations and information processing. The importance of fast synaptic recruitment, action potential initiation and repolarization, and rapid synchronous GABA release by PV+ cells is well established. In contrast, the functional significance of PV+ cell NMDA receptors (NMDARs), which generate relatively slow postsynaptic currents, is unclear. Underlining their importance, several studies implicate PV+ cell NMDAR disruption in impaired network function and circuit pathologies. Here, we show that dendritic NMDARs underlie supralinear integration of feedback excitation from local pyramidal neurons onto mouse CA1 PV+ cells. Furthermore, by incorporating NMDARs at feedback connections onto PV+ cells in spiking networks, we show that these receptors enable cooperative recruitment of PV+ interneurons, strengthening and stabilising principal cell assemblies. Failure of this phenomenon provides a parsimonious explanation for cognitive and sensory gating deficits in pathologies with impaired PV+ NMDAR signalling.

neuroscience↗

bioRxiv2018

Analog closed-loop modulation of hippocampal pyramidal cells dissociates gamma frequency and amplitude

Gamma-band oscillations are implicated in modulation of attention and integration of sensory information. The finding that cross-regional coherence varies with task and performance suggests a role for gamma oscillations in flexible communication among anatomically connected brain areas. How networks become entrained is incompletely understood. Specifically, it is unclear how the spectral and temporal characteristics of network oscillations can be altered on rapid timescales needed for efficient communication. We use closed-loop optogenetic modulation of principal cell excitability to interrogate the dynamical properties of hippocampal oscillations. Gamma frequency and amplitude can be modulated bi-directionally, and dissociated, by phase-advancing or delaying optogenetic feedback to pyramidal cells. Closed-loop modulation alters the synchrony rather than average frequency of action potentials, in principle avoiding disruption of population rate-coding of information. Modulation of phasic excitatory currents in principal neurons is sufficient to manipulate oscillations, suggesting that feed-forward excitation of pyramidal cells has an important role in determining oscillatory dynamics and the ability of networks to couple with one another.

neuroscience↗