NIHRDataInsights/HRCSData huggingface dataset:
Title: Interpretation of guidance cues by the actin-binding protein drebrin to direct collective neuronal migration
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Neuronal migration is a fundamental constituent of normal brain development, yet remains poorly understood. Its failure underlies many congenital disorders so there is a clear need for investigation into how it is controlled. It is a complex process requiring cells to respond to environmental guidance cues and translate them into motile changes. Our focus will be tangential neuronal migration, in which neurons extend a leading process to their target which the cell body follows. This is a key feature of the developing oculomotor system, the model used here. Our extensive preliminary data point to a central role for the actin-binding protein, drebrin, in forming leading processes, coordinating the motile response to extracellular signals and ensuring the collective migration of neuronal pools. drebrin is also important beyond development; its loss is linked to degenerative conditions of old age, particularly dementia. A better understanding of its role is thus critical for improving quality of life from birth right through to old age. We will examine the function of drebrin and its interaction with both axon guidance receptors and intracellular effector proteins. In ovo electroporation will be used to examine the role of drebrin - and the effect of specific mutations - in neuronal migration. This technique will be enhanced by using genetic tools to control the timing and level of drebrin expression and enable us to dissect its involvement at discrete stages e.g. initiation of migration compared to choice of trajectory. These studies on the intact brain will be complemented by fluorescent live cell imaging to analyse drebrin kinetics within neurites in vitro. Such data will reveal how molecular interactions produce macroscopic behavioural changes in cells at individual and population levels. The interactions of drebrin with guidance receptors and adapter proteins will be confirmed biochemically using co-immunoprecipitation and Western blotting.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: A novel role for the chemokine lymphotactin and its receptor in trigeminal sensitisation
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Several chemokines have recently been identified within the nervous system, where they have been shown to have diverse effects, regulating neuronal development, neuroinflammation and synaptic transmission, and have been implicated in the initiation and maintenance of exaggerated pain states. Recent work in Prof Boissonade's laboratory provides the first demonstration that the relatively little-studied chemokine, lymphotactin (XCL1), and its receptor (XCR1) are expressed in the nervous system. The overall aim of this study is therefore to characterise for the first time the expression of XCL1 and XCR1 in the trigeminal system, and to investigate their putative role in hyperexcitability, signalling and trigeminal pain. We will characterise XCR1 and XCL1 expression in normal rat trigeminal tissues (tooth pulp, lingual nerve, trigeminal ganglion and trigeminal nucleus) and identify specific structures demonstrating expression, eg neuronal structures, immune cells and glial cells. We will also determine in our established animal models, whether XCR1 and XCL1 expression are altered by nerve injury or inflammation and establish any correlation with the development of hyperexcitability. We will examine the expression of the XCL1 and XCR1 in human tooth pulps and injured nerves using our unique archives of human tissues from patients with known clinical pain histories. This will enable us to establish whether expression at sites of nerve injury or inflammation correlates with the presence of pain. The above will be achieved using a range of immunocytochemical and image analysis techniques. Using an in-vitro trigeminal slice preparation we will determine whether XCL1 activates intracellular signalling molecules, eg p38 and ERK, in the trigeminal nucleus within specific cell types, eg astrocytes, microglia or neurones, and probe the cellular mechanisms of XCL1-induced activation.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Oxidative stress induced regulation of synaptic growth in the nervous system - dissection of genetic and cellular mechanisms
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Oxidative stress is a hallmark of ageing and neurodegenerative diseases. This proposal seeks to investigate how reactive oxygen species (ROS) impinge on the nervous system, specifically the growth of synaptic terminals, where increases in ROS unexpectedly cause overgrowth. Using the genetically tractable fruitfly, drosophila, as a model we will address key issues. 1. We will determine the effects of different sources of oxidative stress on presynaptic terminals, at late larval neuromuscular junction (NMJ) and in the central nervous system (CNS) as well as postsynaptic dendritic arbors of central neurons. 2. We found that different sources of ROS, cytoplasmic versus mitochondrial, induce synaptic terminal growth via different Jun-N-terminal Kinase (JNK) pathway components and will now identify the relevant upstream components and downstream transcriptional codes. 3. We will establish cellular processes and underlying mechanisms mediating ROS-induced synaptic growth, investigating the role of autophagy and associated genes. We will use genetics to identify interactions between genes and recently developed mosaic and intersectional expression systems to target expression of dominant negative, fluorophore tagged and RNAi transgenes to specific neurons. We will visualize pre- and postsynaptic terminals by immunofluorescence antibody staining and targeted expression of fluorophore tagged reporter constructs, imaging these with point and field scanning confocal systems Live imaging will inform us about changes to the dynamics of ROS induced growth, T.E.M. about the role of autophagy. We will apply 3-D reconstruction algorithms for quantitative analysis of complex branched dendrites. Given the high conservation of the signaling pathways and cellular processes associated with oxidative stress, this proposal is likely to identify new candidate genes as potential targets for therapeutic strategies aimed at ameliorating the effects of ageing in the nervous system.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Do all oligodendrogenic stem cells age at the same rate?
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Loss of myelin sheaths from CNS axons is a prominent histological finding in the aging 'healthy' and diseased brain. This is related to the age-associated decline in the ability of the CNS to generate new cells to replace lost myelin. The main source of replacement cells is the oligodendrocyte progenitor cells (OPCs) that are widely distributed throughout the CNS. These cells are generated during the perinatal period and although capable of self-renewal nevertheless undergo age-associated changes, with an increasing fraction of them becoming senescent. This reduces myelin maintenance in the aged brain. Recently, neural stem cells located in the subependymal zone have been identified as an additional source of OPCs and ultimately new (re)myelinating oligodendrocytes in the brain. Since these niche-derived OPCs are generated from stem cells we hypothesise that they will be less adversely affected than the adult parenchymal OPCs. In this project, we will use transgenic mice (crossing Rosa26-EYFP and GFAP-CreErt2 mouse lines) to specifically label subependymal zone stem cells and their progeny, including OPCs. Based on this technology and using a battery of in vivo and in vitro assays, we will compare the properties (cell cycle kinetics, migration potential, remyelination potential) of the constantly generated niche-derived and potentially young OPCs with the more numerous peri-natally generated ageing OPCs. Importantly, the comparison between young and aged progenitors of the same lineage performed in the context of both the young and aged brain will allow the dissection of cell-intrinsic and extrinsic factors implicated in the age-related changes in the properties of OPCs. This project will advance our knowledge on the oligodendrogenic potential of adult neurogenic areas and will identify molecular pathways involved in the effects of ageing in the overall brain oligodendrogenic machinery.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Targeted drug delivery to neurons and glia using light- and field-sensitive microcapsules
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Targeted drug delivery has been a high-priority issue in both experimental and translational biomedical research. We propose to develop a novel methodology of highly-targeted, time-controlled delivery of signalling molecules into nerve and glial cells. To achieve this, we will exploit a recent breakthrough in bionanotechnology (co-authored by one of the Applicants) focusing on fabrication of microscopic biocapsules with pre-designed properties and intra-lumen load. Firstly, we will develop protocols of synthetic microcapsulation to optimise geometry and mechanical properties of microcapsules for successful delivery in cultures and organised tissue. Secondly, we will adjust microcapsule sensitivity to magnetic field and light (using established nanoparticle insertion technologies) to enable capsule navigation and laser-triggered payload release in situ. Thirdly, we will establish experimental protocols for loading, tracking and laser-triggered opening of microcapsules in neuron-glia cultures and acute brain slices using established one- and two-photon excitation laser scanning microscopy techniques. Specialised hardware will be designed to navigate magnetic field-sensitive microcapsules in cultures and slices under a microscope. Fourthly, we will combine these methods with patch-clamp electrophysiology and fluorescence imaging to optimise efficiency, dynamic range and kinetics of capsular payload release. Finally, we will test the proposed methodology in neurobiological environment, by monitoring physiological effects of drug application in real time for common neurobiological tasks: (a) extracellular delivery of neurotransmitter, (b) targeted ligand delivery inside neurons, (b) targeted toxin delivery inside glia. Achieving the proposal's objectives will not only pave the way for a new dimension of experimental probing in neurobiology, but also lay the foundation for clinically relevant applications in drug delivery.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: Bilateral BBSRC-FAPESP: Amelioration of the autonomic imbalances of old age with exercise - exploring the molecular and physiological mechanisms
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The reasons why homeostatic systems deteriorate in the elderly is not well understood. However, there is growing evidence for autonomic nervous system dysfunction in a wide range of chronic conditions of old age. Whilst it is known that age and a sedentary lifestyle are risk factors in the development of autonomic imbalance, there is evidence in humans that physical activity can both prevent or delay its development, and ameliorate the condition once established. We hypothesise that the development and/or maintenance of autonomic imbalance is associated with changes in the expression of a large number of genes in the brain that, through their activity, form one or more functional networks that can be influenced by both lifestyle factors (eg. physical activity) and age. In order to test this hypothesis, will identify putative networks by carrying out detailed and comprehensive transcriptome analysis on brain regions involved in the regulation of autonomic outflow. Expression will be compared in normal animals (Wistar Kyoto rats), and in animals with a genetic predisposition to autonomic imbalance characterised by the development of hypertension and loss of heart rate variability (Spontaneously Hypertensive rats). We will compare gene expression in young and old animals, and we will ask if expression is altered by physical exercise, initiated either before or after the onset of hypertension. Using bioinformatic tools, we will construct putative pathways and networks that may govern these processes, and we will identify nodal genes with many connections. We will then test whether these hubs are important physiologically by altering their expression using in vivo somatic gene transfer approaches. This will be followed by robust physiological analysis (arterial blood pressure, heart rate/period, spontaneous cardiac baroreceptor reflex gain) that will reveal the roles of these hub genes in network stability and their response to external cues such as exercise training.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: A genetic approach to the study of the neuroprotective role of cysteine string protein during normal ageing
Funder: Biotechnology and Biological Sciences Research Council

Abstract: We will use genetic approaches to study the basic mechanisms underlying the neuroprotective function of cysteine string protein (CSP) during normal ageing. This synaptic chaperone protein has an essential physiological role in preventing presynaptic neurodegeneration. This function was known to be conserved from flies to man and our recent work has shown it is also conserved in Caenorhabditis elegans based on study of mutants alleles of the CSP orthologue dnj-14. Elucidation of the basic mechanisms and regulators of the CSP-dependent pathway in worms could illuminate general aspects of normal neuroprotective pathways. C. elegans is an ideal model for such an investigation due to its simple organisation and the ease of genetic manipulation. In addition, genetic and regulatory factors that influence ageing are conserved from worm to mammals. Mechanistic insight into how CSP exerts its neuroprotective role and why CSP absence results in neurodegeneration is limited although recent work has implicated interactions with the synaptic protein SNAP-25. This proposal aims to use the genetic power of C. elegans to test SNAP-25 (RIC-4 in the worm) as a potential target for CSP action. Biochemical analyses will be used to assess RIC-4 levels and ubiquitination in dnj-14 mutant strains and an epistasis analysis will be carried out by crossing the dnj-14 mutant with ric-4 mutants and other control worm lines. We will aim to identify regulators of the neurodegeneration pathway triggered by knock-out of the CSP orthologue DNJ-14. This will be based on mutagenesis of dnj-14 mutant strains that express GFP in all neurons. Following mutagenesis the whole genomes of selected mutants and the parent strain will be sequenced. Following bioinformatic analysis candidate mutations that act as suppressors of the dnj-14 mutant phenotype (loss of fluorescent neurons and reduced life-span) will be characterised using genetic approaches to confirm the functional significance of the mutated gene.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Shuttling through the Blood Brain Barrier using Carbon Nanoneedles
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Targeted drug delivery across the blood brain barrier (BBB) to the central nervous system is a large challenge for the treatment of neurological disorders. drug delivery systems that cross the BBB without disrupting it would also have many applications for brain cancer diagnostic and other diseases, e.g. Alzheimer's disease. There still remains a pressing need for improved nanoparticle delivery systems to target drugs across the BBB. Functionalized carbon nanotubes (CNTs) are receiving increasing attention as vehicles for the delivery of diagnostic and therapeutic agents. The delivery properties of CNTs derive from the potential to deliver a drug by heterogeneously functionalizing their surface and also their ability to traverse the plasma membrane, promoting the cellular uptake of small molecules. However, there is essentially no mechanistic data at the level of individual CNTs to characterise their fundamental interaction with endothelial cells or how this interaction is modified by CNT surface chemistry and targeting to specific BBB receptors such as transferrin receptor or low density lipoprotein receptor-related protein 1. This project aims towards the evaluation of the BBB penetration capacity of novel f-CNT carriers using an in vitro BBB model employing porcine brain endothelial cells (PBECs) chosen due to its high transendothelial electrical resistance when cultured as a monolayer without astrocytes (>300 ohm.cm2) and good expression of the most important drug efflux transporters and P-glycoprotein. f-CNT will be fluorescently labelled or conjugated to a radioactive tracer to allow for qualitative and quantitative measurement of uptake or BBB permeation. Insights into these mechanisms will be essential to guide the development of CNT vectors to deliver drugs across the BBB to the central nervous system for the treatment of neurological disorders.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: Investigating the role of CD4+ T cells in CNS remyelination
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Ageing can profoundly impair neurological function with devastating biological consequences. Remyelination is a natural regenerative process that replaces damaged myelin to maintain insulation and protection of long axons. The efficiency of remyelination declines with age however underlying mechanisms are poorly understood. CD4+ T cells have been shown to support CNS processes such as learning memory and repair however the role of CD4+ T cells in remyelination has received little attention. One study showed that remyelination is impaired in the absence of CD4+ T cells. However, since that study in 2003, our understanding of CD4+ T cell phenotype and function has expanded dramatically to include a wide range of subsets that are functionally diverse and many are reciprocally antagonistic. Furthermore, ageing impairs CD4+ T cell responses but affects subsets differently. We hypothesise that CD4+ T cell subsets exert differential effects on remyelination and that aged-induced T cell immunosenescence impairs CNS remyelination. To test this we have developed a novel organotypic brain slice (OBS) model of murine hippocampal remyelination. Using transgenic reporter mice we will differentiate Th1, Th2, Th17 and Treg subsets and co-culture purified T cells with remyelinating OBS. We will determine the effect of T cell subsets on remyelination by quantifying remyelination in 3-D using a novel algorithm we have developed. We will investigate the effect of T cell ageing on remyelination by differentiating T cell subsets from young and aged mice and co-culturing these cells with remyelinating OBS. Finally we will increase the complexity of our technology by co-culturing remyelinating OBS with CNS antigen-specific T cells of each subset, mixed with myeloid cells, as would be more representative of the in vivo scenario. Together, these studies will provide insight to the influence of T cells in CNS repair and will develop further, novel technology to study neuroimmune interaction.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Determining the role of M cells in TSE agent neuroinvasion from the intestine
Funder: Biotechnology and Biological Sciences Research Council

Abstract: After oral exposure many TSE agents replicate first on follicular dendritic cells (FDC) in the gut-associated lymphoid tissues (GALT) as they spread from the site exposure (intestine) to the brain (termed, neuroinvasion). For TSE agents to replicate on FDC in the GALT after ingestion of a contaminated meal they must first cross the gut epithelium. However, the precise cellular mechanism by which TSE agents are conveyed into Peyer's patches is not known. Within the Peyer's patch epithelium are M cells, unique epithelial cells specialized for the transepithelial transport of particles. M cells are plausible sites of TSE agent transport across the gut epithelium, but definitive demonstration in vivo is lacking. This is important, as several M cell-independent mechanisms have also been proposed. A thorough understanding of the early stages of TSE pathogenesis in the GALT is crucial to determine the factors that influence the risk of infection and the identification of cellular and molecular targets for intervention. Using unique in vivo models will be used to definitively determine the role of M cells in TSE agent neuroinvasion from the intestine and test the hypothesis that M cells play a crucial role in oral TSE pathogenesis. Objective 1 tests the hypothesis that in the specific absence of M cells, TSE agent accumulation in the GALT is blocked and neuroinvasion impaired. Objective 2 aims to determine whether inflammatory stimuli that enhance M cell differentiation also enhance TSE agent uptake. Objective 3 will determine whether inflammation-mediated villous M cell expansion exacerbates TSE pathogenesis by facilitating systemic dissemination and neuroinvasion from tissues other than Peyer's patches. These data will help predict how inflammation and co-fection with gastrointestinal pathogens may influence TSE pathogenesis. This study will provide important missing data on how orally-acquired TSE agents infect the GALT during steady-state and inflammatory conditions.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Targeted cell-specific deletion of Kir4.1 channels to determine their functions in oligodendrocytes
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The project is basic research to examine the fundamental importance of the Kir4.1 potassium channel subtype in oligodendrocytes. We have demonstrated that oligodendrocytes strongly express Kir4.1 and that there is severe oligodendrocyte dysfunction following global ablation of Kir4.1 in conventional knock out (KO) mice. However, the precise function of Kir4.1 channels in oligodendrocytes are unknown and it is not clear why they are so critically important for myelin-formation. Our studies highlight the crucial importance of Kir4.1 in oligodendrocytes, but in the global Kir4.1 KO mouse the effects on oligoderocytes represent developmental defects, and it is not possible to distinguish between direct and indirect effects. Determination of the precise functions of Kir4.1 in oligodendrocytes requires targeted disruption of these channels. We will use tamoxifen-inducible CreERT2/loxP gene targeting to create the first inducible KO mice in which the Kir4.1 gene is specifically disrupted in oligodendrocytes and their progenitors. Generation of these mouse strains will enable us to control the temporal and cell-specific expression of Kir4.1, with which we will be able to make major advances in our understanding of the functions of oligodendrocyte Kir4.1 in the developing and ageing brain.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL