Life Sciences Commons^™

Mechanisms Of Diapause And Cold Tolerance In The Colorado Potato Beetle, Jacqueline E. Lebenzon

Electronic Thesis and Dissertation Repository

Many temperate insects enter diapause (a state of dormancy) and enhance their cold tolerance to survive the winter. During diapause, the Colorado potato beetle (CPB, Leptinotarsa decemlineata, Coleoptera: Chrysomelidae) stops developing, lowers its metabolism, and changes its physiology to avoid freezing. The extent to which diapause confers cold tolerance in CPB is currently unknown. In my thesis, I used CPB to improve our understanding of the mechanisms underlying metabolic suppression during diapause and cellular protection at sub-zero temperatures in insects. First, I used RNA-sequencing (RNA-seq) to compare gene expression in two metabolically important tissues (the fat body and flight muscle) …

Hibernation Is Super Complex: Dynamics Of Electron Transport System Supercomplexes, Amalie J. Hutchinson

Electronic Thesis and Dissertation Repository

The electron transport system complexes form supercomplexes (SCs) within mitochondrial membranes, perhaps increasing respiratory capacity or reducing reactive oxygen species production. My project aimed to determine the abundance, composition, and stability of SCs in a hibernator. Hibernators have dynamic metabolisms that change greatly during the winter. I isolated mitochondria from rats and thirteen-lined ground squirrels (TLGS) in different hibernation states and measured mitochondrial respiration. I extracted mitochondrial proteins using two detergents of different strengths, and quantified SC abundance using 2-dimensional gel electrophoresis and immunoblotting. Rats had fewer SCs than TLGS. SCs are dynamic in hibernation and the complex III composition …

Metabolic Regulation During Extraembryonic Endoderm Differentiation, Mohamed Gatie

Electronic Thesis and Dissertation Repository

The inner cell mass undergoes orchestrated cellular divisions resulting in the formation of the epiblast (EPI) and primitive endoderm (PrE). Understanding the process of cell fate specification is crucial to appreciate the intricacies of proper embryonic development. While the mouse embryo is an excellent model, limitations do exist with number, technical challenges, and accessibility, therefore, in my thesis I employed two cell-based models to recapitulate the EPI-PrE fate in vitro. Many signaling pathways have been implicated in this lineage decision, metabolism and its downstream products have been recently regarded as a driver of lineage commitment. Using various biochemical, molecular, …

Characterizing The Effects Of Pyrroloquinoline Quinone (Pqq) Supplementation On Skeletal Muscle Mitochondrial Function And Myogenesis During Oxidative Stress And Iugr., Allyson J. Wood

Electronic Thesis and Dissertation Repository

Intrauterine growth restriction (IUGR) affects 10-15% of births and is associated with placental insufficiency (PI), resulting in fetal oxidative stress (OS). This OS is a factor in the predisposition to postnatal noncommunicable disease (NCD) of which muscle mitochondrial dysfunctional is a key aspect. Pyrroloquinoline quinone (PQQ), an antioxidant-like compound, is capable of OS reduction and promotes mitochondrial function, though limited research has focused on its effects in in utero skeletal muscle. This study sought to investigate the impact of in vitro H₂O₂, a model of OS, and an in vivo model of OS associated IUGR, with …

Phenotypic And Metabolic Plasticity In Canine Cellular Reprogramming, Ian C. Tobias

Electronic Thesis and Dissertation Repository

Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) derived from large mammals reproduce few characteristics displayed by rodent or human counterparts. This is complicated by the inherent plasticity of mammalian ESC/iPSC cultures that resemble a variety of developmental stages including the naïve and primed pluripotent states. Defining the extrinsic signals that modulate the developmental identity of canine ESC/iPSC (i.e. primed versus naïve) will improve knowledge integration with more sophisticated rodent and primate research. In this thesis, I sought to determine if manipulation of the culture environment can promote nuclear and metabolic reprogramming of canine cell lines towards a …

Structure And Function Of Stomatin-Like Protein 2, Safee Mian

Electronic Thesis and Dissertation Repository

Stomatin-like protein 2 (SLP-2), a member of the SPFH superfamily, is a mitochondrial inner membrane protein required for optimal mitochondrial respiration. SLP-2 binds to the important mitochondrial phospholipid cardiolipin (CL) and has been proposed to mediate formation of CL-enriched microdomains that would foster respiratory chain supercomplex (RCS) formation and stability. However, little is known about how SLP-2 structure facilitates its cellular function. The goal of this thesis was to elucidate if and how SLP-2 oligomerizes and by what means does it bind CL.

Biophysical analysis of the expressed SLP-2 SPFH domain, either with or without flanking residues, indicates it to …

Alpha-Synuclein Toxicity Is Caused By Mitochondrial Dysfunction, Michael G. Tauro

Electronic Thesis and Dissertation Repository

Parkinson’s disease (PD) is the second most common neurodegenerative disorder, affecting roughly 1% of the population over the age of sixty years. Alpha-synuclein (aSyn) is a protein implicated in both familial and idiopathic forms of PD, yet despite the wealth of data implicating aSyn as a causative agent in PD, the mechanisms underlying its toxicity remain mostly unknown. Mitochondrial dysfunction is a major hallmark of PD, yet there is only limited evidence linking aSyn toxicity to mitochondrial dysfunction. My study establishes a novel aSyn model in respiring yeast cells, which allows me to explore how aSyn affects mitochondrial homeostasis and …

Investigating The Protective Effects Of Telomerase Reverse Transcriptase On Neuronal Metabolism And Resistance To Amyloid-Beta, Olivia Singh

Electronic Thesis and Dissertation Repository

Maintenance of telomere length during cell division is dependent on the catalytic subunit telomerase reverse transcriptase (TERT), which adds TTAGGG repeats to the ends of chromosomes to prevent telomere shortening during DNA replication. However, non-telomeric roles of TERT have emerged under oxidative stress whereby TERT translocates from the nucleus to the mitochondria and protects against mitochondrial dysfunction through a poorly defined mechanism. A major pathological feature of Alzheimer’s Disease (AD) is the progressive accumulation of amyloid-beta (Aβ) peptide within the cortex and hippocampus. Aβ can directly interfere with mitochondrial respiration and promote mitochondrial dysfunction, ROS production, and neuronal cell death. …

Assisted Reproductive Technologies Disrupt Genomic Imprinting In Human And Mitochondria In Mouse Embryos, Carlee R. White

Electronic Thesis and Dissertation Repository

Infertile couples worldwide use assisted reproductive technologies (ARTs) to help conceive their own biological child. Due to the rising use of ARTs, there is continual emergence of new techniques implemented in human fertility clinics. When treatment is successful, there is an increased risk even within singletons for perinatal complications including preterm birth, intrauterine growth restriction, low and high birth weight and genomic imprinting disorders Beckwith Wiedemann Syndrome, Angelman Syndrome, and Silver-Russel Syndrome. Consequently, there is a need to investigate the effects of these treatments on the manipulated oocyte and preimplantation embryo. To address this, I first analyzed the combined effects …

Temperature-Dependent Alterations Of Brown Adipose Tissue Metabolism During Hibernation In The Thirteen-Lined Ground Squirrel, Ictidomys Tridecemlineatus., Sarah V. Mcfarlane

Electronic Thesis and Dissertation Repository

Brown adipose tissue (BAT) is the major thermogenic tissue in small eutherian mammals. In hibernators, seasonal modifications of BAT are well documented but little is known about its functional regulation during hibernation. BAT metabolism is activated by sympathetic stimulation, so I hypothesized that further regulation at the mitochondrial level, as seen in other hibernator tissues, would be of little advantage. Contrary to my predictions, respiration rates of BAT mitochondria isolated from torpid thirteen-lined ground squirrels were suppressed by up to 62% compared with rates from individuals that aroused to interbout euthermia (IBE), when measured at 37°C. At 10°C, however, these …

Skeletal Muscle Lipid Metabolism And Markers Of Insulin Resistance In Young Male Low Birth Weight Offspring In Combination With A Postnatal Western Diet, Kristyn Pamela Dunlop

Electronic Thesis and Dissertation Repository

Low birth weight offspring are at increased risk for developing metabolic syndrome in later life, specifically its precursor, insulin resistance (IR). Reduced mitochondrial lipid metabolism is implicated in IR pathogenesis, promoting accumulation of acylcarnitines, and potentiating alterations in phosphorylation status of insulin signaling intermediates. While high-energy Western diets are classically implicated in IR progression, the in utero environment was recently highlighted as a major programming mechanism of later life IR. Using a guinea pig model of placental insufficiency, we investigated how an adverse in utero environment impacts later life mitochondrial lipid metabolism and IR progression, as well as its interaction …

Effects Of Growth Temperatures And Elevated Co2 On Respiration Rates In Norway Spruce, Yulia Kroner

Electronic Thesis and Dissertation Repository

Projected increase in growth temperatures and CO₂ may affect carbon balance in Norway spruce (Picea abies), a dominant coniferous species of the boreal forest ecosystem. To examine this, I exposed three-year-old Norway spruce seedlings to six treatments: ambient (400 ppm) and elevated (750 ppm) CO₂ concentrations combined with three growth temperatures: ambient, ambient +4 ^oC, and ambient +8 ^oC. I found that while net growth was generally not affected by growth CO₂ or temperature, leaf nitrogen concentrations were reduced, mortality rates were higher, and needles were shorter and thinner in +8 ^oC …

Are Long Chain Acyl Coas Responsible For Suppression Of Mitochondrial Metabolism In Hibernating 13-Lined Ground Squirrels?, Alex Cooper

Electronic Thesis and Dissertation Repository

I found 44% suppression of succinate-fuelled liver mitochondrial respiration in torpid 13-lined ground squirrels compared to interbout euthermia (IBE). Palmitoyl CoA, predicted to suppress respiration by inhibiting succinate transport at the dicarboxylate transporter (DCT), reduced respiration by ~70%, while butylmalonate, a known inhibitor of the DCT, only inhibited respiration by ~40%. In both cases inhibition of respiration proportionally affected both torpid and IBE mitochondria, suggesting that the DCT is likely not already inhibited in torpid mitochondria. The addition of carnitine, predicted to reverse suppression by facilitating transport of palmitoyl CoA into the mitochondrial matrix, had no rescuing effect on the …

Changes In The Mitochondrial Phosphoproteome During Mammalian Hibernation, Dillon J. Chung

Electronic Thesis and Dissertation Repository

Hibernation in mammals involves a suppression of metabolic rate thereby conserving energy. Many hibernation studies focus on the mitochondria due to their energy-producing role. I hypothesized that this pattern of rapid suppression of liver mitochondrial metabolism with gradual reversal, was related to changes in the phosphorylation state of Ictidomys tridecemlineatus’ mitochondrial proteins in different stages of torpor bouts. Phosphorylation of threonine and serine amino-acid residues (unique to 49, 41 and 47, 38 kDa proteins respectively), detected using immunoblotting, changes seasonally (threonine: summer active > winter, serine: summer active > torpor), whereas no seasonal difference was observed in phosphotyrosine residues. Changes in …