Biology subjects

Whittington, M. A.

Publications and source records attributed to Whittington, M. A..

9 recordsLinked to original sources

bioRxiv2025

Spatial profiling of central nervous system cellular markers in murine breast cancer to brain metastases

BackgroundBrain metastasis occurs in approximately 16% of metastatic breast cancer patients, and incidence is increasing. Patients with breast cancer brain metastases frequently develop neurological problems including cognitive impairments and seizures. We previously demonstrated accumulation of activated microglia around the tumour site in a breast cancer brain metastasis model, which co-localised with spontaneously occurring local field potential events that resembled interictal epileptic discharges. However, the mechanisms underlying these spatially restricted effects on neuronal excitability are poorly understood. MethodsTo better understand the underlying cellular changes and potential mechanisms, we used digital spatial profiling of brain cell subtype-specific markers to chart the spatial organisation of neurons, oligodendrocytes, astrocytes, and microglia in relation to metastatic lesion site. We developed a novel chordline-based approach to analyse these spatial changes in biomarker distribution. ResultsSeveral protein markers associated with proliferation (Ki67), astrocytes (GFAP) and microglia (CD11b, IBA1, CD45, MSR1) were upregulated in the tumour compared to healthy contralateral brain parenchyma. In contrast, some neuroglial markers (MAP2, Neurofilament light, NeuN, S100B and TMEM119) were lower in the lesion compared to normal tissue. ConclusionsOverall, the protein marker changes in the lesion are indicative of neuronal cell loss and changes in the immune microenvironment of the metastatic lesion involving activated microglia, astrocytes and recruitment of peripheral immune cells. Such cellular changes may contribute to the changes in electrical activity resulting from brain metastases observed in mice and patients.

neuroscience↗

bioRxiv2024

Cortical layer-specific and cell type-specific dendritic expression of Arc (Arg 3.1) in an in vitro model of slow-wave sleep

An abundance of evidence shows sleep homeostatically regulates synaptic plasticity and memory consolidation but the underlying molecular regulators of this important function of sleep have been difficult to directly elucidate. Arc is an immediate early gene that has a fundamental role in several aspects of synaptic plasticity including regulation and maintenance. Using a physiological brain slice model of slow-wave sleep, here we show that Arc protein has a characteristic spatial and cell-type specific distribution during persistent sleep-related delta oscillations in neocortical slices, which may correlate with aspects of sleep-dependent regulation of synaptic plasticity in cortical regions. In delta oscillating slices, Arc is highly expressed in layer 2/3 dendrites of the primary and secondary association cortex and this dendritic localisation is specific to intrinsically bursting cells (IB) whose cell bodies are in layer 5. Moreover, Arc immunopositive dendrites are clustered together arranged in a quasi hexagonal arrangement with a spacing of [~] 50 {micro}m between clusters. The cytoarchitectural distribution of Arc across the association cortex has implications for the mechanisms of sleep and for the synaptic homeostasis hypothesis regarding the function of sleep.

neuroscience↗

bioRxiv2020

Nasal respiration is necessary for the emergence of ketamine-induced high frequency network oscillations and behavioral hyperactivity in freely moving rats.

Changes in oscillatory activity are widely reported after subanesthetic ketamine, however their mechanisms of generation are unclear. Here, we tested the hypothesis that nasal respiration underlies the emergence of high-frequency oscillations (130-180 Hz, HFO) and behavioral activation after ketamine in freely moving rats. We found ketamine 20 mg/kg provoked "fast" theta sniffing in rodents which correlated with increased locomotor activity and HFO power in the OB. Bursts of ketamine-dependent HFO were coupled to "fast" theta frequency sniffing. Theta coupling of HFO bursts were also found in the prefrontal cortex and ventral striatum which, although of smaller amplitude, were in phase with OB activity. Haloperidol 1 mg/kg pretreatment prevented ketamine-dependent increases in fast sniffing and instead HFO coupling to slower basal respiration. Consistent with ketamine-dependent HFO being driven by nasal respiration, unilateral naris blockade led to an ipsilateral reduction in ketamine-dependent HFO power compared to the control side. Bilateral nares blockade reduced ketamine-induced hyperactivity and HFO power and frequency. In conclusion, nasal entrainment of ketamine-dependent HFO across cortical and subcortical regions at theta frequencies represents a mechanism of orchestrated neural activity across distinct brain regions. The dense divergent connectivity of the olfactory system serves to broadcast this HFO to limbic areas.

neuroscience↗

bioRxiv2020

Network and synaptic mechanisms underlying high frequency oscillations in the rat and cat olfactory bulb under ketamine-xylazine anesthesia.

High frequency oscillations (HFO) are receiving increased attention for their role in health and disease. Ketamine-dependent HFO have been identified in cortical and subcortical regions in rodents, however, the mechanisms underlying their generation and whether they occur in higher mammals is unclear. Here, we show under ketamine-xylazine anesthesia, classical gamma oscillations diminish and a prominent > 80 Hz oscillation emerges in the olfactory bulb of rats and cats. In cats negligible HFO was observed in the thamalus and visual cortex indicating the OB was a suitable site for further investigation. Simultaneous local field potential and thermocouple recordings demonstrated HFO was dependent on nasal airflow. Silicon probe mapping studies spanning almost the entire dorsal ventral aspect of the OB revealed this rhythm was strongest in ventral areas of the bulb and associated with microcurrent sources about the mitral layer. Pharmacological microinfusion studies revealed HFO was dependent on excitatory-inhibitory synaptic activity, but not gap junctions. Finally, we showed HFO was preserved despite surgical removal of the piriform cortex. We conclude that ketamine-dependent HFO in the OB are driven by nasal airflow and local dendrodendritic interactions. The relevance of our findings to ketamines model of psychosis in awake state are also discussed.

neuroscience↗

bioRxiv2020

Differential contributions of synaptic and intrinsic inhibitory currents to parsing via flexible phase-locking in neural oscillators

Current hypotheses suggest that speech segmentation - the initial division and grouping of the speech stream into candidate phrases, syllables, and phonemes for further linguistic processing - is executed by a hierarchy of oscillators in auditory cortex. Theta (~3-12 Hz) rhythms play a key role by phase-locking to recurring acoustic features marking syllable boundaries. Reliable synchronization to quasi-rhythmic inputs, whose variable frequency can dip below cortical theta frequencies (down to ~1 Hz), requires "flexible" theta oscillators whose underlying neuronal mechanisms remain unknown. Using biophysical computational models, we found that the flexibility of phase-locking in neural oscillators depended on the types of hyperpolarizing currents that paced them. Simulated cortical theta oscillators flexibly phase-locked to slow inputs when these inputs caused both (i) spiking and (ii) the subsequent buildup of outward current sufficient to delay further spiking until the next input. The greatest flexibility in phase-locking arose from a synergistic interaction between intrinsic currents that was not replicated by synaptic currents at similar timescales. Flexibility in phase-locking enabled improved entrainment to speech input, optimal at mid-vocalic channels, which in turn supported syllabic-timescale segmentation through identification of vocalic nuclei. Our results suggest that synaptic and intrinsic inhibition contribute to frequency-restricted and -flexible phase-locking in neural oscillators, respectively. Their differential deployment may enable neural oscillators to play diverse roles, from reliable internal clocking to adaptive segmentation of quasi-regular sensory inputs like speech. Author summaryOscillatory activity in auditory cortex is believed to play an important role in auditory and speech processing. One suggested function of these rhythms is to divide the speech stream into candidate phonemes, syllables, words, and phrases, to be matched with learned linguistic templates. This requires brain rhythms to flexibly synchronize with regular acoustic features of the speech stream. How neuronal circuits implement this task remains unknown. In this study, we explored the contribution of inhibitory currents to flexible phase-locking in neuronal theta oscillators, believed to perform initial syllabic segmentation. We found that a combination of specific intrinsic inhibitory currents at multiple timescales, present in a large class of cortical neurons, enabled exceptionally flexible phase-locking, which could be used to precisely segment speech by identifying vowels at mid-syllable. This suggests that the cells exhibiting these currents are a key component in the brains auditory and speech processing architecture.

neuroscience↗

bioRxiv2019

Flexible reset and entrainment of delta oscillations in primate primary auditory cortex: modeling and experiment

Salient auditory stimuli typically exhibit rhythmic temporal patterns. A growing body of evidence suggests that, in primary auditory cortex (A1), attention is associated with entrainment of delta rhythms (1 - 4 Hz) by these auditory stimuli. It is thought that this entrainment involves phase reset of ongoing spontaneous oscillations in A1 by thalamus matrix afferents, but precise mechanisms are unknown. Furthermore, naturalistic stimuli can vary widely in terms of their rhythmicity: some cycles can be longer than others and frequency can drift over time. It is not clear how the auditory system accommodates this natural variability. We show that in rhesus macaque monkey A1 in vivo, bottom-up gamma (40 Hz) click trains influence ongoing spontaneous delta rhythms by inducing an initial delta-timescale transient response, followed by entrainment to gamma and suppression of delta. We then construct a computational model to reproduce this effect, showing that transient thalamus matrix activation can reset A1 delta oscillations by directly activating deep (layer 5) IB cells, promoting bursting, and beginning a new delta cycle. In contrast, long duration gamma-rhythmic input stimuli induce a steady-state containing entrainment of superficial RS and FS cells at gamma, and suppression of delta oscillations. This suppression is achieved in the model by two complementary pathways. First, long-duration thalamus matrix input causes IB cells to switch from bursting to sparse firing, which disrupts the IB bursts associated with delta. Second, thalamus core input activates deep FS cells (by way of layer 4), which fire at gamma frequency and actively inhibit the delta oscillator. Together, these two fundamental operations of reset and suppression can respectively advance and delay the phase of the delta oscillator, allowing it to follow rhythms exhibiting the type of variability found in the natural environment. We discuss these findings in relation to functional implications for speech processing.\n\nAuthor summaryNeurons organize their firing into synchronous, rhythmic patterns. These neural oscillations have been shown to entrain to rhythmic stimuli in the external world, such as patterns of speech or patterns of movement. By entraining to a particular input stimulus, these oscillations are thought to help us attend to that stimulus and to exclude others. To understand how this synchronization emerges, we constructed a physiologically detailed mathematical model of the primary auditory cortex. By fitting this model to a variety of experimental data, we suggest fundamental mechanisms by which neurons of the auditory cortex can synchronize their activity to rhythmic external stimuli. This result will be useful for understanding the mechanism and limitations of oscillatory entrainment, which are thought to underlie the processing of naturalistic auditory inputs like speech or music. Furthermore, this model, though simplified, was shown to generalize and reproduce a wide range of experimental results, and can thus be used as a starting point for building more complex models of auditory cortex.

neuroscience↗

bioRxiv2019

Metastatic breast cancer cells induce altered microglial morphology and electrical excitability in vivo

BackgroundAn emerging problem in the treatment of breast cancer is the increasing incidence of metastases to the brain. Metastatic brain tumours are incurable and can cause epileptic seizures and cognitive impairment, so better understanding of this niche, and the cellular mechanisms, is urgently required. Microglia are the resident brain macrophage population, becoming \"activated\" by neuronal injury, eliciting an inflammatory response. Microglia promote proliferation, angiogenesis and invasion in brain tumours and metastases. However, the mechanisms underlying microglial involvement appear complex and better models are required to improve understanding of function.\n\nMethodsHere, we sought to address this need by developing a model to study metastatic breast cancer cell-microglial interactions using intravital imaging combined with ex vivo electrophysiology. We implanted an optical window on the parietal bone to facilitate observation of cellular behaviour in situ in the outer cortex of heterozygous Cx3cr1GFP/+ mice. Results: We detected GFP-expressing microglia in Cx3cr1GFP/+ mice up to 350 {micro}m below the window without significant loss of resolution. When DsRed-expressing metastatic MDA-MB-231 breast cancer cells were implanted in Matrigel under the optical window, significant accumulation of activated microglia around invading tumour cells could be observed. This inflammatory response resulted in significant cortical disorganisation and aberrant spontaneously-occurring local field potential spike events around the metastatic site.\n\nConclusionsThese data suggest that peritumoral microglial activation and accumulation may play a critical role in local tissue changes underpinning aberrant cortical activity, which offers a possible mechanism for the disrupted cognitive performance and seizures seen in patients with metastatic breast cancer.

cancer biology↗

bioRxiv2018

Insoluble Aβ overexpression in an App knock-in mouse model alters microstructure and gamma oscillations in the prefrontal cortex, and social and anxiety-related behaviours

We studied two new App knock-in mice models of Alzheimers disease (AppNL-F and AppNL-G-F), which generate elevated levels of A{beta}40 and A{beta}42 without the confounds associated with APP overexpression. This enabled us to assess changes in anxiety-related and social behaviours, and neural alterations potentially underlying such changes, driven specifically by A{beta} accumulation. AppNL-G-F knock-in mice exhibited subtle deficits in tasks assessing social memory, but not in social motivation tasks. In anxiety-assessing tasks, AppNL-G-F knock-in mice exhibited: 1) increased thigmotaxis in the Open Field (OF), yet; 2) reduced closed-arm, and increased open-arm, time in the Elevated Plus Maze (EPM). Their ostensibly-anxiogenic OF profile, yet ostensibly-anxiolytic EPM profile, could hint at altered cortical mechanisms affecting decision-making (e.g. disinhibition), rather than simple core deficits in emotional motivation. Consistent with this possibility, alterations in microstructure, glutamatergic-dependent gamma oscillations, and glutamatergic gene expression were all observed in the prefrontal cortex, but not the amygdala, of AppNL-G-F knock-in mice. Thus, insoluble A{beta} overexpression drives prefrontal cortical alterations, potentially underlying changes in social and anxiety-related behavioural tasks.\n\nHighlightsO_LIAppNL-G-F KI mice displayed differing anxiety behaviours in two tests of anxiety.\nC_LIO_LIPrefrontal gamma was no longer NMDA-receptor dependent in AppNL-G-F KI mice.\nC_LIO_LIPrefrontal expression of Grin2b was reduced in AppNL-G-F KI mice.\nC_LIO_LIDTI found structural alterations in the hippocampus and prefrontal cortex in AppNL-G-F KI mice.\nC_LI

neuroscience↗

bioRxiv2018

Cross-fostering in rodents causes region-specific alterations in entorhinal cortical gamma rhythms associated with NMDA receptor dysfunction

There has recently been a large increase in the number of children placed in foster care in the United States and Europe. While this is the least worst scenario for those with a lack of appropriate biological care, it is recognised that these children are exposed to major stressors correlated with behavioural changes, particularly in the realm of social cognition into adulthood. Here we model foster care in rodents: rat pups are removed from their biological mother and placed with a non-genetically related dam. This prevented the entorhinal cortex from generating patterns of gamma rhythms required for normal parahippocampal function relevant to social interaction. These changes correlated with a reduction in NMDA receptor-mediated excitation, and changes in parvalbumin expression in interneurons. These data suggest that early life care delivered by a non-biological parent may disrupt social behaviour but, in contrast, generate neurobiological changes antagonistic to those currently associated with psychosis.\n\nSignificance StatementCross fostering is an effective approach for delineating the effect of environment from genetic influences upon behavior. This involves removal of pups from one mother and transfer to another lactating dam. This manipulation is considered as a mild form of early life stress, producing neurobehavioral changes such as alterations in social interaction. We demonstrate that cross fostering produces changes in the ability of cortical microcircuits to generate oscillatory rhythms, in particular the gamma rhythm, in brain regions important for social cognition. This reduction in gamma rhythmogenesis is related to a reduction in synaptic drive provided by the NMDA receptor. One implication of this work is that the modulation of NMDA receptors offers a potential therapeutic strategy for disorders involving impaired sociability.

neuroscience↗