Engaging with Qualitative Data Analysis: The Metaphor of Looking at Data like a Landscape to Be Explored

“Looking at data like a landscape to be explored” is the metaphor used by Susanne Friese in her new book entitled Qualitative Data Analysis with ATLAS.ti (2014). In it, Friese successfully guides new and experienced researchers to use ATLAS.ti as a computer-assisted tool to engage in qualitative data analysis. The book transcends from being a dry reading to become a companion for those who are living the journey of using different sources of data, mapping them and then integrated them into their analysis

FMRP sustains presynaptic function via control of activity-dependent bulk endocytosis

Synaptic vesicle (SV) recycling is essential for the maintenance of neurotransmission, with a number of neurodevelopmental disorders linked to defects in this process. Fragile X syndrome (FXS) results from a loss of fragile X mental retardation protein (FMRP) encoded by the FMR1 gene. Hyperexcitability of neuronal circuits is a key feature of FXS, therefore we investigated whether SV recycling was affected by the absence of FMRP during increased neuronal activity. We revealed that primary neuronal cultures from male Fmr1 knock-out (KO) rats display a specific defect in activity-dependent bulk endocytosis (ADBE). ADBE is dominant during intense neuronal activity, and this defect resulted in an inability of Fmr1 KO neurons to sustain SV recycling during trains of high-frequency stimulation. Using a molecular replacement strategy, we also revealed that a human FMRP mutant that cannot bind BK channels failed to correct ADBE dysfunction in KO neurons, however this dysfunction was corrected by BK channel agonists. Therefore, FMRP performs a key role in sustaining neurotransmitter release via selective control of ADBE, suggesting intervention via this endocytosis mode may correct the hyperexcitability observed in FXS. SIGNIFICANCE STATEMENT Loss of fragile X mental retardation protein (FMRP) results in fragile X syndrome (FXS), however whether its loss has a direct role in neurotransmitter release remains a matter of debate. We demonstrate that neurons lacking FMRP display a specific defect in a mechanism that sustains neurotransmitter release during intense neuronal firing, called activity-dependent bulk endocytosis (ADBE). This discovery provides key insights into mechanisms of brain communication that occur because of loss of FMRP function. Importantly it also reveals ADBE as a potential therapeutic target to correct the circuit hyperexcitability observed in FXS

Whole school singing at two Canadian independent secondary schools: "it is the life-blood of our school"

The purpose of this dissertation was to investigate the phenomenon of whole-school communal singing at two Canadian independent secondary schools. Research questions included: What does the practice of whole-school communal singing look like, how is it experienced by participants, and how has the practice been initiated and maintained? Previous research on whole school singing at the secondary school level is scant. This suited an exploratory, phenomenological research methodology for the present study. Pascale’s (2005) two aesthetics of singing provided the theoretical framework. The idea that choral singing can be approached through a broader lens than is currently practiced in typical choral education contexts helped to characterize whole school singing as a communal singing practice. Research literature explored prior to data analysis focused on communal singing, defined as participatory singing by everyone in a non-choir community. Examples included crowd singing at sports games, at protest marches, and in churches. Communal singing in North America was more popular in the early 20th century than today, which may explain its rareness in contemporary secondary schools. Data were collected through interviews with 17 current and former students, faculty and administrators at two schools. Analysis was conducted using thematic analysis (Braun & Clarke, 2006), supported by NVivo software, and resulted in five overarching themes. The first was that all the research participants expressed a strong positive regard for the practice of whole school singing; the second was that communal singing may contribute to student belongingness. The third overarching theme was that communal singing appears to mediate emotions and may contribute to student wellness; fourth, that the approach taken to whole school singing at the two schools prioritizes full participation over achieving aesthetic qualities typically espoused by performance choirs. The fifth theme was that whole school singing at the secondary school level is not easy to initiate and maintain, but requires specific leadership, intention, and strategy in order to create a fully participative, engaging, and joyful experience in a secondary school context. Post data analysis, findings were compared and contrasted with those from related research. Recommendations are provided for educators who may wish to consider incorporating communal singing into school life

Synaptic vesicle recycling in preclinical models of intellectual disability, autism spectrum disorder and epilepsy

The development of the central nervous system is dysregulated in neurodevelopmental disorders such as intellectual disability, autism spectrum disorder, and epilepsy. These three disorders have different clinical features, yet there is high comorbidity between them. They can be difficult to study due to their highly complex aetiologies, however there are various monogenic diseases that can cause all of them, including SYNGAP1 haploinsufficiency where the synaptic guanosine triphosphatase (GTPase)-activating protein (SYNGAP) protein levels are highly reduced; Fragile X syndrome where the fragile X mental retardation protein (FMRP) is no longer translated; and DNM1 epileptic encephalopathy where mutations in the Dynamin1 gene alter the protein function. These monogenic conditions are synaptopathies as the proteins affected play important roles in synapse stability and neurotransmission. Because of the high comorbidity between these disorders, it is hypothesised that there may be a common mechanism underlying them. We hypothesise that a deficit in presynaptic vesicle recycling may be part of a common mechanism underlying intellectual disability, autism spectrum disorder, and epilepsy especially in SYNGAP1 haploinsufficiency, Fragile X syndrome, and DNM1 epileptic encephalopathy. Using various fluorescent presynaptic activity reporters including synaptic pHluorins, tetramethylrhodamine dextran and calcium dyes to compare presynaptic activity in in vitro models of these monogenic conditions, we found differences in synaptic vesicle (SV) endocytosis in the genetically altered conditions compared to wildtype controls. We observed various SV endocytosis defects in clathrin-mediated endocytosis (CME) or activity-dependent bulk endocytosis (ADBE) in our models. We observed enhanced CME in SynGAP1 KO mouse hippocampal neurons. This enhanced SV endocytosis was accompanied by decreased SV cargo on the plasma membrane. Rat SynGAP1 KO hippocampal neurons did not display enhanced SV endocytosis, nor did neurons with the GTPase-activating (GAP) domain of SynGAP deleted. This was perhaps due to the altered time course of development between these rodent species. In mouse and rat models of Fragile X syndrome, CME was not altered compared to wildtype controls. However, in a rat model, we observed fewer nerve terminals undergoing ADBE which is the dominant SV endocytosis mode during elevated neuronal activity. De novo epileptic encephalopathy-associated mutations in DNM1 had differential effects on SV recycling through both CME and ADBE. Mouse hippocampal neurons overexpressing Dyn1R237W, Dyn1I289F and Dyn1H396D all showed less CME compared to overexpression of Dyn1WT. Moreover, fewer nerve terminals overexpressing Dyn1H396D were found to undergo ADBE. We also found that a large-conductance potassium (BK) channel opener can accelerate clathrin-mediated endocytosis and thus may be able to rescue the impaired SV endocytosis caused by these mutants. Although there is not yet a common underlying pathway at the presynaptic level between these conditions, SV recycling dysfunction is present across all of these models. Furthermore, we propose an axis of pathophysiology model where optimal SV endocytosis is required for optimised neural performance. We propose that either decreased or increased SV endocytosis can lead to the synaptic dysfunction observed in these models

A novel synaptopathy-defective synaptic vesicle protein trafficking in the mutant CHMP2B mouse model of frontotemporal dementia

Mutations in the ESCRT-III subunit CHMP2B cause frontotemporal dementia (FTD) and lead to impaired endolysosomal trafficking and lysosomal storage pathology in neurons. We investigated the effect of mutant CHMP2B on synaptic pathology, as ESCRT function was recently implicated in the degradation of synaptic vesicle (SV) proteins. We report here that expression of C-terminally truncated mutant CHMP2B results in a novel synaptopathy. This unique synaptic pathology is characterised by selective retention of presynaptic SV trafficking proteins in aged mutant CHMP2B transgenic mice, despite significant loss of postsynaptic proteins. Furthermore, ultrastructural analysis of primary cortical cultures from transgenic CHMP2B mice revealed a significant increase in the number of presynaptic endosomes, while neurons expressing mutant CHMP2B display defective SV recycling and alterations to functional SV pools. Therefore, we reveal how mutations in CHMP2B affect specific presynaptic proteins and SV recycling, identifying CHMP2B FTD as a novel synaptopathy. This novel synaptopathic mechanism of impaired SV physiology may be a key early event in multiple forms of FTD, since proteins that mediate the most common genetic forms of FTD all localise at the presynapse

Mino Bimaadiziwin Homebuilder Program’s Impact on Sustainable Livelihoods Among Youth in Garden Hill and Wasagamack First Nations: An Evaluative Study

The Mino Bimaadiziwin Homebuilders postsecondary education pilot project built Indigenous youth capacity and houses in two remote Anishinini reserves—Garden Hill and Wasagamack. To evaluate this community-led project, a sustainable livelihood assessment holistically measured the impact on 45 of the 70 (64%) Homebuilder students and the community. The community benefited by gaining three culturally appropriate houses built from local lumber and employment opportunities for Anishinini instructors. A longitudinal survey found five of the six livelihood assets improved statistically and significantly, including satisfaction with social relationships, cultural awareness, income and ability to pay bills, housing safety, and human development. Students reported better relations with their families and neighbourhood. Most (85%) of the 70 Homebuilder students earned postsecondary certificates either in forestry, homebuilding or both while obtaining a training stipend, which elevated their incomes. These positive outcomes occurred despite project underfunding, the COVID-19 pandemic lockdown, climate change events, and inequitable housing policies under the Indian Act. Based on this project’s success, we recommend investing in Indigenous-led postsecondary education in community homebuilding projects. However, to attain equitable housing and human rights, a plan is needed to overturn the Indian Act, which keeps Indigenous people as “wards of the state” and their land in trust

Reversal of cell, circuit and seizure phenotypes in a mouse model of DNM1epileptic encephalopathy

Dynamin-1 is a large GTPase with an obligatory role in synaptic vesicle endocytosis at mammalian nerve terminals. Heterozygous missense mutations in the dynamin-1 gene (DNM1) cause a novel form of epileptic encephalopathy, with pathogenic mutations clustering within regions required for its essential GTPase activity. We reveal the most prevalent pathogenic DNM1 mutation, R237W, disrupts dynamin-1 enzyme activity and endocytosis when overexpressed in central neurons. To determine how this mutation impacted cell, circuit and behavioural function, we generated a mouse carrying the R237W mutation. Neurons from heterozygous mice display dysfunctional endocytosis, in addition to altered excitatory neurotransmission and seizure-like phenotypes. Importantly, these phenotypes are corrected at the cell, circuit and in vivo level by the drug, BMS-204352, which accelerates endocytosis. Here, we demonstrate a credible link between dysfunctional endocytosis and epileptic encephalopathy, and importantly reveal that synaptic vesicle recycling may be a viable therapeutic target for monogenic intractable epilepsies.<br/