Master of Science in Biomedical Engineering (MSBME), Wright State University, 2024, Biomedical Engineering

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease characterized by motor neuron (MN) death resulting in paralysis and eventually death. ALS has greater prevalence in older populations sharing characteristics with aging like muscle weakness and MN type specific degeneration. MNs innervate skeletal muscles and control muscle contraction through their excitability which is altered in both conditions. C-Boutons are a cholinergic, excitatory synaptic input to MNs and have been studied in ALS and aging but have produced inconsistent findings and undesired gaps. We used immunohistochemistry to label mouse lumbar spinal cord and separate MN types. 60x imaging and automated analysis was performed providing robust 3D measurements. Our results presented similar findings between two ALS mutations with differing changes in a third mutation. We also show C-Bouton input with age undergoes sex and MN type specific reductions aligning with age-related weakness. Finally, we identify C-Bouton similarities and differences between ALS and aging.

Committee: Sherif M. Elbasiouny Ph.D. (Advisor); David R. Ladle Ph.D. (Committee Member); Tarun Goswami D.Sc. (Committee Member) Subjects: Aging; Biomedical Engineering; Neurosciences

Master of Science (MS), Wright State University, 2023, Biological Sciences

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that causes the degeneration of motor neurons which leads to loss of motor function and eventual death. Over 20 genes have been implicated in ALS's pathogenesis, one being TARDBP, which codes for TDP-43. TDP-43 mislocalizes from the nucleus, accumulates, and then aggregates in the cytoplasm and is linked to cellular toxicity. We have modeled this aggregation and toxicity of TDP-43 in budding yeast. From genetic screens of human genes and their ability to modulate TDP-43 toxicity, we found 50 human genes that were able to reduce TDP-43 toxicity. 12 of these genes expressed the strongest rescue phenotype. Interestingly, these 12 genes did not lower the protein level or aggregation of TDP-43, nor did they exhibit any protein-protein interactions with TDP-43. We then investigated a possible cellular pathway that could be associated with TDP-43 toxicity and examined how our suppressors were able to reverse its effects. We identified four suppressors that were implicated in the cAMP/PKA pathway, with two of them directly downregulating it. Further analysis revealed that out of these four genes, two led to significantly increased cAMP levels compared to TDP-43 alone. We then studied a downstream target of the cAMP/PKA pathway, stress granule formation. Our observation indicated that TDP-43 not only colocalized with stress granules but also stimulated their formation.

Committee: Shulin Ju Ph.D. (Advisor); Michael Markey Ph.D. (Committee Member); Andrew Voss Ph.D. (Committee Member) Subjects: Biology; Cellular Biology; Molecular Biology

Master of Science in Biomedical Engineering (MSBME), Wright State University, 2023, Biomedical Engineering

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease which targets motoneurons (MNs), yet underlying disease mechanisms are not well understood. Evaluating the intrinsic excitability of MNs in ALS could lead to a better understanding of mechanisms causing neurodegeneration. The SOD1-G93A (SOD) model is the most commonly studied animal model of ALS. However, past studies have shown highly conflicting results on SOD MN excitability. Interestingly, I show that depending on the level of membrane depolarization, SOD MNs show opposite results of both hyper- and hypo-excitability. This reveals that differences in the methodology of measuring excitability can heavily impact the study results. Finally, I have investigated the firing abnormalities leading to hypoexcitability in SOD MNs at high levels of membrane depolarization. Results indicate that these firing abnormalities are due to decreased Kv2.1 channel conductance. Furthermore, these firing abnormalities could be the basis for developing a biomarker which could be used to diagnose ALS earlier.

Committee: Sherif Elbasiouny Ph.D. (Advisor); Keiichiro Susuki M.D., Ph.D. (Committee Member); Jaime Ramirez-Vick Ph.D. (Committee Member) Subjects: Engineering; Neurobiology; Neurosciences; Physiology

Doctor of Philosophy, The Ohio State University, 2023, Neuroscience Graduate Studies Program

Single nucleotide variants (SNVs) in the gene encoding Kinesin Family Member 5A (KIF5A), a neuronal motor protein subunit involved in transport along microtubules, have been associated with amyotrophic lateral sclerosis (ALS). ALS is a rapidly progressive and fatal neurodegenerative disease that primarily affects motor neurons. Numerous ALS-associated KIF5A SNVs are clustered near splice site junctions of the penultimate exon 27 and are predicted to alter the carboxy-terminal (C-term) cargo-binding domain of KIF5A. Mis-splicing of exon 27, resulting in exon exclusion, is proposed to be the mechanism by which these SNVs cause ALS. Whether all KIF5A SNVs proximal to exon 27 result in exon exclusion is unclear. To address this question, we designed an in vitro minigene splicing assay in HEK293 cells which revealed heterogeneous site-specific effects on splicing: only 5´ splice site (5´ss) SNVs resulted in exon skipping. We also quantified splicing in select CRISPR-edited human stem cells differentiated to motor neurons and in neuronal tissues from a 5´ss SNV knock-in mouse (Mouse: c.3005+1G>A; Human homolog: c.3020+1G>A), which showed the same result. Moreover, survival of representative 3´ splice site (3´ss), 5´ss, and truncated C-term (ΔC) variant KIF5A (v-KIF5A) motor neurons was significantly reduced compared to wildtype (WT) motor neurons, and overt morphological changes were apparent. While total KIF5A mRNA levels were comparable across cell lines, total KIF5A protein levels were decreased for v-KIF5A lines, suggesting an impairment of protein synthesis or stability. Thus, despite the heterogeneous effect on RNA splicing, KIF5A SNVs near exon 27 similarly reduce the availability of the KIF5A protein, leading to axonal transport defects and motor neuron pathology.

Committee: Stephen Kolb MD, PhD (Advisor); Michael Kearse PhD (Committee Member); Karin Musier-Forsyth PhD (Committee Member); Arthur Burghes PhD (Committee Member) Subjects: Biochemistry; Chemistry; Experiments; Genetics; Molecular Biology; Molecular Chemistry; Neurobiology; Neurology; Neurosciences; Pharmaceuticals; Pharmacology

Master of Science, The Ohio State University, 2023, Genetic Counseling

Background Although genetic testing for amyotrophic lateral sclerosis (ALS) is widely available and utilized, there is no data about how well patients understand their results. Correct understanding of genetic test results is important because it can influence how and when patients share their result with family members and impact decisions about treatments and presymptomatic testing. Aims The aims of this study were to determine what ALS patients understand about their genetic test results and identify successes and challenges in communication of the key implications of genetic test results. Methods A survey was sent to 10,802 ALS patients on the CDC's National ALS Registry. It included questions about demographic information, the participant's genetic test result, and the participant's understanding of their genetic test result. Comprehension was scored for participants with positive and negative results and was described for participants with VUS results. The survey also included a Likert series about genetic testing experience. Results There were 75 participants in the study. The majority of the cohort was white (97.3%) and had high health literacy. There were 15 participants with a positive result (20%), 38 with a negative result (50.6%), 15 with a VUS (20%), and 7 who did not remember their result (9.3%). The majority of participants with a positive result had high comprehension (60%) or mixed comprehension (27%). The most common misunderstanding in the positive group was that 40% of respondents did not understand that their result explained why they developed ALS. In the group of participants with a negative result, only 8% had high comprehension and 60% had mixed comprehension. The most common misunderstanding in the negative group was that 92% did not understand that the test did not completely rule out a genetic cause for their ALS. The participants with VUS results had high variability in their understanding of their results. There we (open full item for complete abstract)

Committee: Jennifer Roggenbuck (Advisor); Victoria Klee (Committee Member); Dawn Allain (Committee Member) Subjects: Genetics; Neurology

Master of Science, The Ohio State University, 2023, Genetic Counseling

With the increasing knowledge of genetics and the expansion of targeted therapy options, genetic testing for conditions like Amyotrophic Lateral Sclerosis (ALS) is increasing in demand. Since relatively few ALS clinics are staffed by a genetic counselor, non-genetics neurology providers will need to play key roles in genetic test result interpretation, communication, and implementation into patient care. However, physicians across various fields of practice have communicated a lack of confidence in interpreting and communicating uncertain genetic results, a common result for patients with ALS who undergo genetic testing. This questionnaire-based randomized controlled study was conducted to assess whether a genetics educational module could increase non-genetics neurology providers' understanding of and comfort with uncertain genetic testing results in ALS-associated genes. This anonymous online study-questionnaire assessed each participant's baseline genetics knowledge and comfort with interpreting and communicating genetic testing results. Following this, participants were randomly assigned to either the intervention or control arm. The intervention arm received a comprehensive genetic testing educational module, while the control arm received a condensed version that only covered inheritance patterns. The study questionnaire then re-evaluated each participant's genetics knowledge. The questionnaire also assessed their clinical interpretation and comfort level when presented with a hypothetical case vignette featuring an uncertain genetic testing result in an ALS-associated gene. While the results did not reach statistical significance due to a low number of respondents, certain trends were observed. As compared to control arm participants, intervention arm participants had a greater increase in knowledge after receiving the educational module and were more likely to include certain key implications of the case vignette uncertain result in their written interpret (open full item for complete abstract)

Committee: Dawn Allain (Advisor); Adam Quick (Committee Chair); Victoria Klee (Committee Member) Subjects: Adult Education; Genetics; Health Education

Master of Science (MS), Wright State University, 2022, Physiology and Neuroscience

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease. To date, there are no significant disease-modifying treatments, and one limiting factor in treatment is the amount of time it takes for a patient to receive a diagnosis of ALS. This study examined multiple mouse models before symptom onset to help identify early changes in a reflex circuit of ALS mice. Dorsal root stimulation of the sacral spinal cord in multiple models ALS mouse models showed changes in the resulting ventral root compound action potential amplitude, latency, and ability to maintain synaptic depression. These data also suggest that a mouse model of TDP43 inclusions is fundamentally different in its network properties than that of SOD1 ALS mutant mice. While these data suggest changes in motor neuron excitability may play a contributing factor, there are likely other synaptic changes involved, but further work needs to be done to verify this.

Committee: Sherif M. Elbasiouny Ph.D., P.E., P.Eng. (Advisor); David Ladle Ph.D. (Committee Member); Mark Rich M.D., Ph.D. (Committee Member) Subjects: Neurosciences

Master of Science (MS), Wright State University, 2022, Biological Sciences

ALS is a neurodegenerative disease characterized by degeneration of upper and lower motor neurons in the brain and spinal cord leading to progressive paralysis and ultimately death. Perturbations in RNA metabolism and RNA binding proteins have emerged as underlying defects in ALS pathogenesis. Matrin-3 is a multifunctional RNA binding protein that has been linked to familial and sporadic ALS. Matrin-3 is normally found in the nucleus, but mutations in the gene cause mislocalization of the protein from the nucleus into the cytoplasm of neuronal cells where it forms protein aggregates. In this study, we show that over-expressing human MATR3 in the budding yeast, Saccharomyces cerevisiae, results in cellular toxicity and cytoplasmic aggregation, recapitulating phenotypes of mutant Matrin-3 in mammalian models and patients. We tested Matrin-3 with other ALS-associated RBPs and identified human genes that rescue cells from Matrin-3-induced growth defects without altering protein aggregation patterns.

Committee: Shulin Ju Ph.D. (Advisor); Labib Rouhana Ph.D. (Committee Member); Quan Zhong Ph.D. (Committee Member) Subjects: Biology; Biomedical Research

Doctor of Philosophy (PhD), Wright State University, 2021, Biomedical Sciences PhD

Amyotrophic lateral sclerosis (ALS) is the most common motorneuron (MN) disease in adulthood. ALS is hallmarked by the progressive loss of MNs in the brain, brainstem, and spinal cord. Many hypotheses to explain the pathogenesis of ALS have been explored, but the exact mechanisms underlying the development of this disease remain unknown. However, abnormalities in MN excitability and glutamate excitotoxicity are the most widely studied. For decades, researchers have examined MN excitability in ALS, but the current literature is inconsistent, showing evidence of hyperexcitability, hypoexcitability, or no change in excitability of MNs in ALS. Many of these studies also focus solely on the excitability of individual MNs, rather than the spinal MN network, whose output collectively drives muscle activity. Using electrophysiology intracellular and ventral root recordings in SOD1-G93AHigh-Copy (SOD) mice, the standard rodent model of ALS, at symptom onset, we demonstrate evidence of both hypo- and hyperexcitability in ALS, whereby disease mechanisms change MN excitability in one direction and compensatory mechanisms alter MN excitability in the opposite direction. Additionally, we show evidence of a novel mechanism contributing to the development of motor dysfunction in ALS at symptom onset, impaired sensorimotor integration. We also studied the effects of a novel treatment for ALS on MN excitability. In recent years, small-conductance calcium-activated potassium (SK) channels have been implicated in the pathogenesis of ALS. In MNs, these channels mediate the afterhyperpolarization (AHP) and synaptic transmission and plasticity and subsequently regulate MN excitability at the individual and network levels. In SOD mice, these channels are significantly reduced throughout disease progression and early treatment with an SK channel activator, CyPPA, restores these deficits. Early treatment with CyPPA also prolongs the survival and motor function of SOD mice. Our results de (open full item for complete abstract)

Committee: Sherif M. Elbasiouny Ph.D. (Advisor); Khalid M. Elased Pharm.D., Ph.D. (Committee Member); Lynn K. Hartzler Ph.D. (Committee Member); Mark M. Rich M.D., Ph.D. (Committee Member); Keiichiro Susuki M.D., Ph.D. (Committee Member) Subjects: Biomedical Research; Cellular Biology; Neurosciences; Physiology

Master of Science (MS), Wright State University, 2020, Physiology and Neuroscience

Protocol development and optimization are vital in the scientific method process. By having accurate protocols, one can properly assess the characteristics of their animal model for any given experiment. One animal newly adopted in our lab was the novel regulatable nuclear localization sequence (rNLS) mouse model. This novel mouse model displays symptoms of Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal dementia (FTD), after the accumulation of the hTDP-43 (TAR DNA-binding protein 43) aggregate in the central nervous system. The expression of this protein occurs after the removal of deoxycycline from the mouse's food source. Once the removal of the drug, this activates a tetracycline-controlled activation system, which causes expression of hTDP-43. The ability to control the expression of hTDP-43 provides the uniqueness to this ALS/FTD mouse model allowing researchers to study these fatal neurodegenerative diseases at various time points in the mouse's timeline. In this thesis, three different studies were conduct that either developed or optimized protocols to assess characteristics of this novel rNLS mouse. The first study investigates the development of cognitive behavioural tasks designed assess working memory and learning in this mouse model. These behavioural tasks are the Y-maze, the NOR (NOR) and the Holeboard tests. In addition to developing behavioural task protocols suited for our mouse model, a comparison was also done between WT (WT) and NS (NS) rNLS mouse, which are mice part of the rNLS colony that do not have the bigenic mutation expressing ALS/FTD, to determine if NS rNLS could be used as control, the results confirm they can be used. In the second study, the Y-maze protocol was utilized in assessing short-term working-memory of rNLS mice 3 and 5 weeks off doxycycline. This preliminary study shows evidence of cognitive deficits in this mouse model as well as provides credibility to our developed protocol. The third study compares the int (open full item for complete abstract)

Committee: Sherif M. Elbasiouny Ph.D., P.E. (Advisor); Keiichiro Susuki M.D., Ph.D. (Committee Member); Adrian M. Corbett Ph.D. (Committee Member); Barry Milligan Ph.D. (Other) Subjects: Neurosciences

Master of Science (MS), Wright State University, 2020, Anatomy

Amyotrophic Lateral Sclerosis is a fatal, progressive, neurodegenerative disease. There currently is no cure for the disease and limited treatment options have modest effects. A hallmark of the disease is motoneuron degeneration. Within the membrane of motoneurons are small-conductance calcium-activated potassium channels. SK Channels function to mediate medium afterhyperpolarization period of an action potential and have substantial influence on the firing rate of motoneurons. SK channels, specifically SK2 and SK3 isoforms, have been shown through immunohistochemistry to be affected in the SOD1G93A mouse model. The goal of this study was to investigate whether neonatal administration of CyPPA, a potent SK2 and SK3 activator, had effects on the clustering of SK2 and SK3 channels in the SOD1G93A mouse model. To explore this, immunohistochemistry was performed at postnatal day 21 and day 90 and determined that CyPPA had influence on the clustering profile of both SK channel isoforms in wild-type and SOD1G93A.

Committee: Sherif Elbasiouny Ph.D., P.E. (Advisor); David R. Ladle Ph.D. (Committee Member); Keiichiro Susuki M.D., Ph.D. (Committee Member) Subjects: Neurosciences

PhD, University of Cincinnati, 2020, Medicine: Neuroscience/Medical Science Scholars Interdisciplinary

Respiratory failure is the leading cause of death in in amyotrophic lateral sclerosis (ALS) patients and spinal cord injury patients. Therefore, it is important to identify neural substrates that may be targeted to improve breathing following disease and injury. We show that a class of ipsilaterally projecting, excitatory interneurons located in the brainstem and spinal cord – V2a neurons – play key roles in controlling respiratory muscle activity in health and disease. We used chemogenetic approaches to increase or decrease V2a excitability in healthy mice and following disease or injury. First, we showed that silencing V2a neurons in neonatal mice caused slow and irregular breathing. However, silencing V2a neurons in adult mice did not alter the regularity of respiration and actually increased breathing frequency, suggesting that V2a neurons play different roles in controlling breathing at different stages of development. V2a neurons also pattern respiratory muscle activity. Our lab has previously shown that increasing V2a excitability activates accessory respiratory muscles at rest in healthy mice. Surprisingly, we show that silencing V2a neurons also activated accessory respiratory muscles. These data suggest that two types of V2a neurons exist: Type I V2a neurons activate accessory respiratory muscles at rest whereas Type II V2a neurons prevent their activation when they are not needed. Moreover, altering the excitability of just cervical spinal neurons using viral strategies suggests that these two V2a subtypes are both located in the cervical spinal cord. The effect of increasing and decreasing V2a excitability on accessory respiratory muscle activity was also tested in ALS model mice. Increasing or decreasing V2a excitability activates accessory respiratory muscles throughout disease progression in SOD1(G93A) ALS model mice. Finally, we demonstrate that increasing V2a excitability promotes recovery of diaphragm function following a high level C2 hemisection (open full item for complete abstract)

Committee: Mark Baccei Ph.D. (Committee Chair); Steven Crone Ph.D. (Committee Chair); Warren Alilain Ph.D. (Committee Member); Steve Davidson Ph.D. (Committee Member); Timothy Weaver Ph.D. (Committee Member) Subjects: Neurology

Master of Science (MS), Wright State University, 2019, Microbiology and Immunology

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motor neuron function. There is no cure for ALS, but studies of genes implicated in the disease continue to provide necessary insight for generating therapeutic targets. Both inherited and sporadic forms of ALS have been found to contain mutations of the nuclear RNA-binding protein FUS, which is mislocalized into cytoplasmic aggregates in ALS motor neurons. To better understand the underlying pathological mechanisms, an established yeast model which recapitulates the mislocalization, aggregation, and cytotoxicity of FUS-ALS has been an indispensable tool. Genetic screens performed using the model enabled the identification of two strong suppressors of FUS toxicity: TAF and NKAPD. Through interchanging structurally similar domains, we elucidate the domains of TAF that are required for rescue. Though rescue by TAF is not through FUS, we report here that NKAPD rescues FUS toxicity by acting on FUS directly. We find that rescue of FUS toxicity is achieved through differing but equally effective approaches.

Committee: Shulin Ju Ph.D. (Advisor); Quan Zhong Ph.D. (Committee Member); Barbara Hull Ph.D. (Committee Member) Subjects: Biology

Doctor of Philosophy (PhD), Wright State University, 2019, Biomedical Sciences PhD

Amyotrophic Lateral Scleroses (ALS) is a neurodegenerative disease characterized by the degeneration of upper and lower motor neurons in the brain and spinal cord leading to progressive paralysis and ultimately death within 5 years of symptom onset. Only two drugs are approved by the Food and Drug Administration (FDA) for treating ALS that slow the disease progression by about 3 months. Mutations in the gene Fused in Sarcoma (FUS) cause inherited forms of ALS. FUS is a nuclear, multifunctional RNA-binding protein (RBP) involved in multiple RNA metabolic pathways. Mutations in FUS cause mislocalization of the protein from the nucleus to cytoplasm, where it forms inclusion that colocalize with stress granules. Over-expression of human FUS in the budding yeast, Saccharomyces cerevisiae, results in cellular toxicity, cytoplasmic aggregation and localization to stress granules, recapitulating phenotypes of mutant FUS in mammalian models and patients. In recent years, perturbations in RNA metabolism and RNA binding proteins has emerged as an underlying defect in ALS pathogenesis. FUS is one of the first RBPs linked to ALS, and modeling FUS toxicity in yeast will enhance our understanding of how RBPs contribute to neuronal toxicity in ALS. Using a yeast model of FUS toxicity, we designed and completed a genetic screen to identify human genes that suppress FUS induced toxicity. Enrichment analysis of the suppressor genes showed an over representation of genes with RNA binding function and ribonucleoprotein complex localization. A subset of the suppressors physically interact with FUS and colocalize with FUS aggregates. We focused on the FUS suppressor TATA-Box Binding Protein Associated Factor (TAF) for further study since TAF has been found in a complex with FUS and has a similar function and structure to FUS. We tested TAF against three other toxic neurodegenerative disease proteins; TAF suppressed toxicity of FUS and not the others. TAF did not reduce FUS pr (open full item for complete abstract)

Committee: Shulin Ju Ph.D. (Advisor); Paula Bubulya Ph.D. (Committee Member); Weiwen Long Ph.D. (Committee Member); Mark Rich M.D., Ph.D. (Committee Member); Quan Zhong Ph.D. (Committee Member) Subjects: Molecular Biology

Master of Science (MS), Wright State University, 2018, Biological Sciences

Optineurin, an autophagy adaptor protein genetically linked to Normal Tension Glaucoma and Amyotrophic Lateral Sclerosis, has been found in the pathological inclusions of various other neurodegenerative disorders, supporting an important role of optineurin in neurodegeneration. Using yeast as a model, we found that overexpression of optineurin drastically enhances the toxicity of the Parkinson's disease-causing protein, alpha-synuclein. Considering the conserved protein-protein interaction between optineurin and Ypt1, a yeast suppressor of the toxicity of both optineurin and alpha-synuclein, we hypothesize that cellular targets of optineurin underlie in the cytotoxicity and the enhancer effect. Using genome-wide yeast two-hybrid screens, we identified 97 yeast interacting proteins of optineurin and systematically tested their modifier effect on the toxicity of optineurin and alpha-synuclein. Given the neuroprotective effect of the mammalian homologs of Ypt1, the convergent pathways of the identified modifiers may represent conserved cellular perturbations induced by optineurin overexpression in higher eukaryotes.

Committee: Quan Zhong Ph.D. (Advisor); Mill Miller Ph.D. (Committee Member); Paula Bubulya Ph.D. (Committee Member) Subjects: Cellular Biology; Genetics; Neurosciences

Master of Science, The Ohio State University, 2018, Genetic Counseling

Background: Although there is an increase of clinicians offering ALS genetic testing, the practice of offering genetic testing in the management of patients with ALS is currently inconsistent without consensus ALS genetic testing guidelines. Methods: We distributed a survey to clinicians that are members of the North East ALS Consortium (NEALS). The survey was comprised of multiple choice questions and a 9 item Likert scale that assessed clinicians' attitudes of the offering of genetic testing. Results: The survey was email to 255 members of the North East ALS Consortium (NEALS); 80 responded (participant response rate = 31.4%). Approximately half (n=59/121, 48.8%) of NEALS sites were represented by at least one response. The majority of respondents (73.9%, 48/65) stated they would be more likely to offer genetic testing in their practice if there were consensus guidelines to direct the offer of ALS genetic testing, with 26.2% (17/65) not more likely to offer genetic testing. The majority of respondents (47/65, 72.3%) indicated patients who reported a family history of ALS; 57% (37/65) indicated patients with a family history of dementia or other neurodegenerative condition, 53.9% (35/65) indicated patients who have a young age of onset (50.8% (33/65) indicated patients who requested genetic testing (and a minority of respondents (24/65, 37%) indicated all ALS patients. Clinicians were specifically asked to indicate when genetic testing is offered to sALS patients, in their practice: 52.3% (34/65) indicated sALS patients with a family history of dementia, 46.2% (30/65) indicated sALS patients with early onset ALS, report and 37.0% (24/65) indicated all sALS patients. A mean attitude score was calculated for each respondent; the average attitude score of the respondents was 2.23 (with a possible range of 0 to 10, with 0 being the most positive), indicating the overall attitude score corresponded to a favorable perception. The association between attitude score (open full item for complete abstract)

Committee: Jennifer Roggenbuck (Advisor) Subjects: Biology; Genetics; Health Care; Health Care Management; Medicine; Neurology

Master of Science (MS), Wright State University, 2017, Physiology and Neuroscience

Amyotrophic Lateral Sclerosis (ALS) is a fatal, adult-onset progressive degenerative motor neuron disease that is characterized by muscle atrophy and weakness due to the loss of upper and lower motor neurons. Average survival time for individuals diagnosed with the disease is three to five years; currently there is no cure and only one drug approved by the Food and Administration (FDA). Scientists have proposed various theories in order to solve the mystery which surrounds ALS. One of these theories hypothesizes how hyperexcitability and excitotoxicity leads to the death of motor neurons. In this study, we will address ways of combatting the effects of hyperexcitability as well as excitotoxicity by targeting a specific channel type. The channels in question are small conductance calcium activated potassium channels (SK channels). We chose to target these channels because they directly affect the medium after-hyperpolarization (mAHP) of the cell which controls firing rate. We postulate that SK channels are being altered in such a way that cell firing rate has been increased, leading to phenotypes associated with the disease such as abnormal excitability, mitochondrial dysfunction, axonal loss motor impairment, muscle atrophy as well as excitotoxicity, thus leading to the spread of motor neuron death. Upon administration with a specific SK channel activator in the form of CyPPA; improvements in motor function and survival were found. These improvements suggest that SK channels are indeed viable drug targets and specific SK channel activators may be treatment options for individuals suffering from ALS.

Committee: Sherif Elbasiouny Ph.D., P.E. (Committee Chair); Mark Rich M.D., Ph.D. (Committee Member); Keiichiro Susuki M.D., Ph.D. (Committee Member) Subjects: Behavioral Sciences; Neurobiology; Neurosciences; Physiology

Master of Science (MS), Wright State University, 2016, Biological Sciences

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative motor neuron disease that causes progressive paralysis and death by asphyxiation. There is no cure or effective treatment; however, previous research has identified several genes that appear related to the pathology of ALS. When mutated, these genes result in proteins that gain toxic functions and disrupt normal cellular processes. Fused in Sarcoma (hFUS) is a human transcription factor in the nucleus that binds to DNA and RNA. Mutations in hFUS are associated with both familial and sporadic cases of ALS, frontotemporal lobar degeneration (FTLD), and cancer. In ALS and FTLD, hFUS is mislocalized to the cytosol where it interacts with stress granules and forms aggregates. This aggregation and cytotoxicity has been previously studied in budding yeast; however, study in fission yeast may provide unique information. Fission yeast has several genetic advantages over budding yeast for modeling mammalian cell biology, such as 43% of genes contain introns and they posess a similar alternative splicing mechanism. Mammalian and fission yeast cells also both contain microRNA as well as similar cell growth cycles. In this project, I established a fission yeast model of hFUS and showed that hFUS is toxic when overexpressed in fission yeast. Both localization to the nucleus and mislocalization to the cytosol occurred during overexpression of hFUS. In addition, fission yeast homologues to previously identified budding yeast toxicity suppression proteins were able to suppress hFUS toxicity, suggesting the suppression mechanism is conserved.

Committee: Shulin Ju Ph.D. (Advisor); Labib Rouhana Ph.D. (Committee Member); Barbara Hull Ph.D. (Committee Member) Subjects: Cellular Biology

Master of Science (MS), Wright State University, 2016, Physiology and Neuroscience

Amyotrophic lateral sclerosis (ALS) is a devastating neuromuscular disease that currently has no cure and extremely limited treatment options. The specific mechanisms that underlie motoneuron degeneration and death, which are classical features of this disease, are mostly unknown. This thesis tests the hypothesis that small-conductance calcium-activated potassium channels (SK) may be downregulated in ALS motoneurons, as suggested by computational modelling. SK channel expression was measured in spinal alpha-motoneuron cell bodies or somata of wildtype (WT) and mutant (mt) SOD1-G93A mice, a transgenic animal model of ALS. Quantitative immunohistochemical analysis of the developmental expression of SK channel isoforms SK2 and SK3 at various postnatal time points was performed to assess the effects of motoneuron degeneration on the level and/or pattern of protein expression on the somata of lower lumbar motor nuclei. Results indicate that the selective expression of SK3 may be gradually reduced over development in WT and mutant SOD1 mice but is affected by disease pathogenesis. In addition, SK channels appear to be clustered in both WT and mutant SOD1 motoneurons throughout development. However, SK clusters appear to be significantly smaller in mutant SOD1 motoneurons compared to their WT littermates. These changes indicate that the activity of SK channels, which regulate the firing rate of motoneurons, may be affected in ALS.

Committee: Sherif Elbasiouny Ph.D., P.E., P.Eng. (Committee Chair); David Ladle Ph.D. (Committee Member); Keiichiro Susuki Ph.D. (Committee Member) Subjects: Neurobiology; Neurosciences; Physiology

Master of Science (MS), Wright State University, 2016, Biological Sciences

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disorder that occurs due to the death of motor neurons and leads to paralysis and death within three to five years after symptoms present (Byrne et al., 2013). Superoxide Dismutase 1 (SOD1) was first identified to be associated with ALS in 1993. The objective of this study is to determine which proteins interact with wild type and mutant SOD1 and find any similarities or differences between them. ALS is attributed to a gain of toxicity, therefore abnormal protein interactions in mutant SOD1 are important. The results of this study will provide insight on the protein-protein interactions of SOD1, as well as how important these interactions are in association with ALS. Initially, the plan was to use yeast two-hybrid screening (Y2H) to identify the protein-protein interactions, then confirm the interactions with a pull down assay (immunoprecipitation). However, the Y2H was unable to obtain results. Instead, a combination of a pull-down assay and mass spectrometry were used to identify protein-protein interactions. Fifty one proteins were identified to interact exclusively with wild type SOD1 and thirteen proteins interacted with both wild type and A4V SOD1.

Committee: Shulin Ju Ph.D. (Advisor); Mill Miller Ph.D. (Committee Member); Paula Bubulya Ph.D. (Committee Member); Quan Zhong Ph.D. (Committee Member) Subjects: Biology; Cellular Biology