Study of the effect of mitochondrial deficiency on the neuronal morphology of the nematode <em>Caenorhabditis elegans</em>

Neurons are highly polarized cells that constantly undergo remodeling of their branches during and after their development. This energy demanding process depends on energy provision from mitochondria via the oxidative phosphorylation (OXPHOS). Although the molecular mechanisms connecting the two are important for understanding pathologies linked to defective neuronal branching due to mitochondrial deficiency, they still remain elusive. In this study, we approached this problem by using as a model a highly branched sensory neuron in Caenorhabditis elegans which demonstrates altered branching pattern in conditions of mitochondrial deficiency. In particular, we show that animals carrying mutations in genes encoding mitochondrial complex I subunits have more dendrites on their sensory neurons compared to wild type animals. The increased outgrowth is not a cell type-specific phenomenon since other types of sensory neurons demonstrate ectopic structures due to complex I deficiency, as well. Importantly, the increased branching is not a result of oxidative stress, but it is regulated by a distinct signaling cascade. Specifically, activation of AMP-activated protein kinase AMPK due to mitochondrial deficiency is one of the regulatory events and we show that its downstream target phosphoinositide 3-kinase PI3K participates in the same pathway. Finally, we demonstrated that the increased neuronal arborizations displayed by OXPHOS mutants are independent of their longevity. Overall, our findings reveal a signaling pathway which regulates tightly dendritic branching in conditions of mitochondrial deficiency and contribute to the understanding of more complicated phenomena occurring in relevant neurodegenerative diseases

Lifelong dietary protein restriction induces denervation and skeletal muscle atrophy in mice

© 2024 The Authors. Published by Elsevier Inc.As a widespread global issue, protein deficiency hinders development and optimal growth in offspring. Maternal low-protein diet influences the development of age-related diseases, including sarcopenia, by altering the epigenome and organ structure through potential increase in oxidative stress. However, the long-term effects of lactational protein restriction or postnatal lifelong protein restriction on the neuromuscular system have yet to be elucidated. Our results demonstrated that feeding a normal protein diet after lactational protein restriction did not have significant impacts on the neuromuscular system in later life. In contrast, a lifelong low-protein diet induced a denervation phenotype and led to demyelination in the sciatic nerve, along with an increase in the number of centralised nuclei and in the gene expression of atrogenes at 18 months of age, indicating an induced skeletal muscle atrophy. These changes were accompanied by an increase in proteasome activity in skeletal muscle, with no significant alterations in oxidative stress or mitochondrial dynamics markers in skeletal muscle later in life. Thus, lifelong protein restriction may induce skeletal muscle atrophy through changes in peripheral nerves and neuromuscular junctions, potentially contributing to the early onset or exaggeration of sarcopenia.Biotechnology and Biological Sciences Research Council (BB/P008429/1 & BB/W018276/1. U.E); Turkish Embassy in London; MRC-Versus Arthritis Centre for Integrated research into Musculoskeletal Agein

Guidelines on experimental methods to assess mitochondrial dysfunction in cellular models of neurodegenerative diseases

Neurodegenerative diseases are a spectrum of chronic, debilitating disorders characterised by the progressive degeneration and death of neurons. Mitochondrial dysfunction has been implicated in most neurodegenerative diseases, but in many instances it is unclear whether such dysfunction is a cause or an effect of the underlying pathology, and whether it represents a viable therapeutic target. It is therefore imperative to utilise and optimise cellular models and experimental techniques appropriate to determine the contribution of mitochondrial dysfunction to neurodegenerative disease phenotypes. In this consensus article, we collate details on and discuss pitfalls of existing experimental approaches to assess mitochondrial function in in vitro cellular models of neurodegenerative diseases, including specific protocols for the measurement of oxygen consumption rate in primary neuron cultures, and single-neuron, time-lapse fluorescence imaging of the mitochondrial membrane potential and mitochondrial NAD(P)H. As part of the Cellular Bioenergetics of Neurodegenerative Diseases (CeBioND) consortium ( www.cebiond.org ), we are performing cross-disease analyses to identify common and distinct molecular mechanisms involved in mitochondrial bioenergetic dysfunction in cellular models of Alzheimer's, Parkinson's, and Huntington's diseases. Here we provide detailed guidelines and protocols as standardised across the five collaborating laboratories of the CeBioND consortium, with additional contributions from other experts in the field

Neuronal cell-based high-throughput screen for enhancers of mitochondrial function reveals luteolin as a modulator of mitochondria-endoplasmic reticulum coupling

Background: Mitochondrial dysfunction is a common feature of aging, neurodegeneration, and metabolic diseases. Hence, mitotherapeutics may be valuable disease modifiers for a large number of conditions. In this study, we have set up a large-scale screening platform for mitochondrial-based modulators with promising therapeutic potential. Results: Using differentiated human neuroblastoma cells, we screened 1200 FDA-approved compounds and identified 61 molecules that significantly increased cellular ATP without any cytotoxic effect. Following dose response curve-dependent selection, we identified the flavonoid luteolin as a primary hit. Further validation in neuronal models indicated that luteolin increased mitochondrial respiration in primary neurons, despite not affecting mitochondrial mass, structure, or mitochondria-derived reactive oxygen species. However, we found that luteolin increased contacts between mitochondria and endoplasmic reticulum (ER), contributing to increased mitochondrial calcium (Ca2+) and Ca2+-dependent pyruvate dehydrogenase activity. This signaling pathway likely contributed to the observed effect of luteolin on enhanced mitochondrial complexes I and II activities. Importantly, we observed that increased mitochondrial functions were dependent on the activity of ER Ca2+-releasing channels inositol 1,4,5-trisphosphate receptors (IP3Rs) both in neurons and in isolated synaptosomes. Additionally, luteolin treatment improved mitochondrial and locomotory activities in primary neurons and Caenorhabditis elegans expressing an expanded polyglutamine tract of the huntingtin protein. Conclusion: We provide a new screening platform for drug discovery validated in vitro and ex vivo. In addition, we describe a novel mechanism through which luteolin modulates mitochondrial activity in neuronal models with potential therapeutic validity for treatment of a variety of human diseases

Efficient random codebooks and databases for lossy compression in near-linear time

We examine the compression-complexity trade-off of lossy compression algorithms that are based on a random codebook or a random database. Motivated, in part, by some recent results of Gupta-Verdù-Weissman (GVW) and their underlying connections with the pattern-matching scheme of Kontoyiannis' lossy Lempel-Ziv algorithm, we introduce a non-universal version of the lossy Lempel-Ziv method (termed LLZ), we prove its optimality for memoryless sources, and we compare its performance to that of the GVW divide-and-conquer approach. Experimental results indicate that the GVW approach often yields better compression than LLZ, but at the price of much higher memory requirements. To combine the advantages of both, we introduce a hybrid algorithm (HYB) that utilizes both the divide-andconquer idea of GVW and the single-database structure of LLZ. We show that HYB shares with GVW the exact same ratedistortion performance and implementation complexity, while, like LLZ, requiring much less memory, typically by at least one or two orders of magnitude. Experimental results are also presented, illustrating the performance of all three methods on data generated by simple discrete memoryless sources. In particular, the HYB scheme is shown to outperform existing schemes for the compression of some simple discrete sources with respect to the Hamming distortion criterion. © 2009 IEEE

Lossy Compression in Near-Linear Time via Efficient Random Codebooks and Databases

The compression-complexity trade-off of lossy compression algorithms that are based on a random codebook or a random database is examined. Motivated, in part, by recent results of Gupta-Verd\'{u}-Weissman (GVW) and their underlying connections with the pattern-matching scheme of Kontoyiannis' lossy Lempel-Ziv algorithm, we introduce a non-universal version of the lossy Lempel-Ziv method (termed LLZ). The optimality of LLZ for memoryless sources is established, and its performance is compared to that of the GVW divide-and-conquer approach. Experimental results indicate that the GVW approach often yields better compression than LLZ, but at the price of much higher memory requirements. To combine the advantages of both, we introduce a hybrid algorithm (HYB) that utilizes both the divide-and-conquer idea of GVW and the single-database structure of LLZ. It is proved that HYB shares with GVW the exact same rate-distortion performance and implementation complexity, while, like LLZ, requiring less memory, by a factor which may become unbounded, depending on the choice or the relevant design parameters. Experimental results are also presented, illustrating the performance of all three methods on data generated by simple discrete memoryless sources. In particular, the HYB algorithm is shown to outperform existing schemes for the compression of some simple discrete sources with respect to the Hamming distortion criterion

Neuron-specific proteasome activation exerts cell non-autonomous protection against amyloid-beta (Aβ) proteotoxicity in Caenorhabditis elegans

Proteostasis reinforcement is a promising approach in the design of therapeutic interventions against proteinopathies, including Alzheimer's disease. Understanding how and which parts of the proteostasis network should be enhanced is crucial in developing efficient therapeutic strategies. The ability of specific tissues to induce proteostatic responses in distal ones (cell non-autonomous regulation of proteostasis) is attracting interest. Although the proteasome is a major protein degradation node, nothing is known on its cell non-autonomous regulation. We show that proteasome activation in the nervous system can enhance the proteasome activity in the muscle of Caenorhabditis elegans. Mechanistically, this communication depends on Small Clear Vesicles, with glutamate as one of the neurotransmitters required for the distal regulation. More importantly, we demonstrate that this cell non-autonomous proteasome activation is translated into efficient prevention of amyloid-beta (Αβ)-mediated proteotoxic effects in the muscle of C. elegans but notably not to resistance against oxidative stress. Our in vivo data establish a mechanistic link between neuronal proteasome reinforcement and decreased Aβ proteotoxicity in the muscle. The identified distal communication may have serious implications in the design of therapeutic strategies based on tissue-specific proteasome manipulation

Healthspan improvement and anti-aggregation effects induced by a marine-derived structural proteasome activator

Proteasome activation has been shown to promote cellular and organismal healthspan and to protect against aggregation-related conditions, such as Alzheimer's disease (AD). Various natural compounds have been described for their proteasome activating properties but scarce data exist on marine metabolites that often possess unique chemical structures, exhibiting pronounced bioactivities with novel mechanisms of action. In this study, we have identified for the first time a marine structural proteasome activator, namely (1R,3E,6R,7Z,11S,12S)-dolabella-3,7,18-trien-6,17-olide (DBTO). DBTO activates the 20S proteasome complex in cell-free assays but also in cellulo. Continuous supplementation of human primary fibroblasts with DBTO throughout their cellular lifespan confers an improved healthspan while ameliorated health status is also observed in wild type (wt) Caenorhabditis elegans (C. elegans) nematodes supplemented with DBTO. Furthermore, treatment of various AD nematode models, as well as of human cells of neuronal origin challenged with exogenously added Aβ peptide, with DBTO results in enhanced protection against Aβ-induced proteotoxicity. In total, our results reveal the first structural proteasome activator derived from the marine ecosystem and highlight its potential as a compound that might be used for healthspan maintenance and preventive strategies against proteinopathies, such as AD. © 2022 The Author