Biology subjects

Stout, A.

Publications and source records attributed to Stout, A..

6 recordsLinked to original sources

bioRxiv2026

SEZ6L2 Loss Disrupts Motor Coordination, Cognitive Function, and Synaptic Connectivity

The SEZ6 family, composed of SEZ6, SEZ6L, and SEZ6L2, plays essential roles in neurodevelopment, synaptic organization, and complement regulation. However, the specific contribution of SEZ6L2 to brain function remains largely unexplored. In this study, we provide the first comprehensive behavioral and neurobiological characterization of Sez6l2 knockout (KO) mice and directly compare their phenotype with Sez6 triple knockout (TKO) mice, which lack all three Sez6 family genes. Sez6l2 KO mice exhibit impairments across multiple behavioral domains, including motor coordination, gait, sociability, sensory processing, and goal-directed repetitive behaviors. Several phenotypes, particularly motor deficits, worsen with age. Male Sez6l2 KO mice also demonstrate enhanced fear learning and increased prepulse inhibition, revealing sex-specific alterations in sensorimotor gating. At the synaptic level, Sez6l2 KO mice show reduced dendritic spine length and decreased expression of key postsynaptic proteins suggesting impaired excitatory synaptic connectivity. These structural and molecular abnormalities likely contribute to the observed behavioral deficits. In comparison, Sez6 TKO mice display more severe impairments across most measures. Together, these findings establish SEZ6L2 as a critical and non-redundant regulator of motor, cognitive, and synaptic function and provide mechanistic insight into how dysfunction within the SEZ6 family may contribute to neurodevelopmental and neurodegenerative disorders.

neuroscience↗

bioRxiv2025

Synergetic hallmark knockouts immortalize bovine muscle stem cells for cellular agriculture

Achieving scalable and sustainable production of cultivated meat hinges on developing robust livestock muscle and fat cell lines that can proliferate and differentiate effectively, while meeting regulatory standards and consumer expectations. In this study, bovine satellite cells (BSCs) were immortalized using CRISPR/Cas9 to knockdown PTEN (phosphatase and tensin homolog), TP53 (cellular tumor antigen) and SMAD4 (SMAD [Sma and Mad proteins] family member 4). The resulting cell line, named "CriBSC2," exhibited consistent growth, maintained muscle cell characteristics, and successfully differentiated into multinucleated myotubes after more than 150 cell divisions. In contrast, another cell line, "CriBSC1," achieved immortalization with TP53 knockout alone but lacked differentiation capacity. CriBSC2 were further cultured on gelatin scaffolds to evaluate their anchorage-dependent responses, laying the groundwork for their potential application in tissue engineering for cellular agriculture. Ultimately, CriBSC2 cells demonstrate suitable proliferation and differentiation capabilities crucial for advancing cellular agriculture and future food technologies.

cell biology↗

bioRxiv2025

Sez6L2 autoimmunity induces cerebellar ataxia in mice

Sez6L2 autoantibodies have been reported in patients with subacute cerebellar ataxia, presenting with gait disturbances, frequent falls, slurred speech, and repetitive eye movements. These case studies suggest that autoimmunity against Sez6L2 causes cerebellar damage leading to ataxia. Sez6L2 is a transmembrane protein expressed by most neurons, with the highest levels in the cerebellum. We tested whether immunizing C57BL/6 mice against Sez6L2 could produce an autoimmune response resulting in ataxia symptoms and immune attack on the brain/cerebellum. We found that immunized mice generated significant levels of anti-Sez6L2 antibodies, with IgGs present in the cerebellar parenchyma. Additionally, Sez6L2-immunized mice developed a significant population of Sez6L2-specific T cells targeting two immunodominant epitopes and showed increased CD4+ T cell infiltration into the brain. These mice exhibited mild mobility impairments in open field, wire grid walk, and pole test assays. Our results indicate that an autoimmune response to Sez6L2 in mice can lead to mobility impairments and pathology consistent with human cerebellar ataxia associated with Sez6L2 autoantibodies. This new mouse model should be useful for mechanistic studies on this poorly understood autoimmune disease and for pre-clinical testing of therapeutic strategies targeting the immune system in patients with Sez6L2 antibodies.

neuroscience↗

bioRxiv2023

Hybrid Amyloid Quantum Dot Nanoassemblies to Probe Neuroinflammatory Damage

Various oligomeric species of amyloid-beta have been proposed to play different immunogenic roles in the cellular pathology of Alzheimers Disease. However, investigating the role of a homogenous single oligomeric species has been difficult due to highly dynamic oligomerization and fibril formation kinetics that convert between many species. Here we report the design and construction of a quantum dot mimetic for larger spherical oligomeric amyloid species as an "endogenously" fluorescent proxy for this cytotoxic species to investigate its role in inducing inflammatory and stress response states in neuronal and glial cell types.

neuroscience↗

bioRxiv2021

In vitro and computational analysis of the putative furin cleavage site (RRARS) in the divergent spike protein of the rodent coronavirus AcCoV-JC34 (sub-genus luchacovirus)

The Coronaviridae is a highly diverse virus family, with reservoir hosts in a variety of wildlife species that encompass bats, birds and small mammals, including rodents. Within the taxonomic group alphacoronavirus, certain sub-genera (including the luchacoviruses) have phylogenetically distinct spike proteins, which remain essentially uncharacterized. Using in vitro and computational techniques, we analyzed the spike protein of the rodent coronavirus AcCoV-JC34 from the sub-genus luchacovirus, previously identified in Apodemus chevrieri (Chevriers field mouse). We show that AcCoV-JC34--unlike the other luchacoviruses--has a putative furin cleavage site (FCS) within its spike S1 domain, close to the S1/S2 interface. The pattern of basic amino acids within the AcCoV-JC34 FCS (-RR-R-) is identical to that found in "pre-variant" SARS-CoV-2--which is in itself atypical for an FCS, and suboptimal for furin cleavage. Our analysis shows that, while containing an -RR-R-motif, the AcCoV-JC34 spike "FCS" is not cleaved by furin (unlike for SARS-CoV-2), suggesting the possible presence of a progenitor sequence for viral emergence from a distinct wildlife host.

microbiology↗

bioRxiv2021

Simple and effective serum-free medium for sustained expansion of bovine satellite cells for cell cultured meat

Cell-cultured meat offers the potential for a more sustainable, ethical, resilient, and healthy food system. However, research and development has been hindered by the lack of suitable serum-free media that enable the robust expansion of relevant cells (e.g., muscle satellite cells) over multiple passages. Recently, a low-cost serum-free media (B8) was described for induced pluripotent stem cells. Here, we adapt this media for bovine satellite cells and show that the addition of a single component, recombinant albumin, renders B8 suitable for the long-term expansion of cells without sacrificing myogenicity. We show that this new media (Beefy-9) maintains robust cell growth over the entire culture period tested (seven passages) with an average growth rate of 39 hours per population doubling. Along with demonstrated efficacy for bovine cells, this work provides a promising starting-point for developing serum-free media for cultures from other meat-relevant species. Ultimately, this work offers a promising foundation for escaping the reliance on serum in cultured meat research, thereby accelerating the field.

bioengineering↗