Membrane contact sites and cytoskeleton‐membrane interactions in autophagy

Duckney, Patrick; Wang, Pengwei; Hussey, Patrick J.

doi:10.1002/1873-3468.14414

Cited by 11 publications

(7 citation statements)

References 68 publications

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“…Функциональные митохондриальные нарушения являются взаимосвязанным фактором многочисленных заболеваний, в том числе и в репродуктивной сфере [2]. Данный тезис привел к предположению, что как аномальные метаболические маркеры митохондрий, так и количественные и качественные нарушения функционирования митохондриальных сетей ассоциированы с неблагоприятным репродуктивным прогнозом [3].…”

Section: актуальностьunclassified

Mitochondrial networks of cumulus cells and oocyte quality

Panferov

Tapilskaya²,

Masieva³

et al. 2022

Russian Military Medical Academy Reports

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BACKGROUND: Mitochondria play vital roles in oocyte functions and they are critical indicators of oocyte quality which is important for fertilization and development into viable offspring. Quality-compromised oocytes in which mitochondrial dysfunction plays a significant role are correlated with infertility, developmental disorders and embryo loss. A significant role in the oocytes of cytoplasmic accumulation and a sufficient amount of the mitochondrial pool of oocytes is played by the cumulus cells surrounding it. AIM: to conduct a comparative analysis between the state of mitochondrial networks of cumulus cells and the quality of oocytes. MATERIALS AND METHODS: The study included 22 women aged 28 to 37 years inclusive (mean age 32.3 1.2 years) with tubal infertility. During the assisted reproductive technology procedures, 74 oocytes were obtained. 39 good quality oocytes and their cumulus cells were compared with 35 bad quality oocytes and their cumulus cells respectively. After puncture of ovarian follicles cumulus cells were isolated and stained with fluorescent dye for cell tracing and mitochondrial imaging in vivo. The method of confocal microscopy was used to analyze the three-dimensional organization of mitochondria in 2030 cells of one pool of follicular fluid. The following parameters were used to evaluate the mitochondrial network: fluorescence decay time and fluorescence intensity. RESULTS: The decay time of fluorescence in cumulus cells associated with a good quality oocyte was significantly higher (p = 0.032) than in the case of identification of oocytes with negative signs. CONCLUSION: The structure of mitochondrial networks in cumulus cells correlates (r = 0.76) with the quality of oocytes.

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Section: актуальностьunclassified

Mitochondrial networks of cumulus cells and oocyte quality

Panferov

Tapilskaya²,

Masieva³

et al. 2022

Russian Military Medical Academy Reports

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“…Proteins that act as tethers between organelles facilitate these organelle associations at MCS (Gao et al., 2016; Li et al., 2022; Osterrieder et al., 2017; Wang et al., 2014; White et al., 2020); however, only a few plant MCS tethering proteins have been identified to date. Interactions between organelles play important roles in communication (e.g., exchange of small molecules) and organelle biogenesis (reviewed in Duckney et al., 2022; Oikawa et al., 2019; Perico & Sparkes, 2018). To investigate the ability of a protein to tether two organelles together, tools are required to physically move organelles away from each other.…”

Section: Introductionmentioning

confidence: 99%

Applying Optical Tweezers with TIRF Microscopy to Quantify Physical Interactions Between Organelles in the Plant Endomembrane System

et al. 2023

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Plant organelles are associated with each other through tethering proteins at membrane contact sites (MCS). Methods such as total internal reflection fluorescence (TIRF) optical tweezers allow us to probe organelle interactions in live plant cells. Optical tweezers (focused infrared laser beams) can trap organelles that have a different refractive index to their surrounding medium (cytosol), whilst TIRF allows us to simultaneously image behaviors of organelles in the thin region of cortical cytoplasm. However, few MCS tethering proteins have so far been identified and tested in a quantitative manner. Automated routines (such as setting trapping laser power and controlling the stage speed and distance) mean we can quantify organelle interactions in a repeatable and reproducible manner. Here we outline a series of protocols which describe laser calibrations required to collect robust data sets, generation of fluorescent plant material (Nicotiana tabacum, tobacco), how to set up an automated organelle trapping routine, and how to quantify organelle interactions (particularly organelle interactions with the endoplasmic reticulum). TIRF‐optical tweezers enable quantitative testing of putative tethering proteins to reveal their role in plant organelle associations at MCS. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Microscope system set‐up and stabilityBasic Protocol 2: Generation of transiently expressed fluorescent tobacco tissue by Agrobacterium‐mediated infiltrationBasic Protocol 3: Setting up an automated organelle trapping routineBasic Protocol 4: Quantifying organelle interactions

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“…In plants, members of the Networked (NET) family possess actin-binding capacity, localize to different compartments and have been shown to participate in membrane contact site formation (Duckney et al, 2022). Actin filaments and microtubules represent the molecular tracks for moving organelles and molecules within a cell (Vale, 2003).…”

Section: Introductionmentioning

confidence: 99%

Networked proteins redundantly interact with VAP27 and RABG3 to regulate membrane tethering at the vacuole and beyond

Kaiser,

Mehlhorn,

Miranda

et al. 2023

Preprint

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Biological processes in eukaryotes depend on the spatio-temporal compartmentalization of their cells. Integrity and positioning of organelles on the other hand rely on the organization of the actin cytoskeleton. Previously, it has been shown that changes of the plants largest organelle, the vacuole, depends on a functional actin organization. The connection between actin filaments and the vacuole is established by the family of Networked (NET) 4 proteins and, consequently, altering NET4 abundance impacts vacuolar morphology. However, the precise regulatory mechanism is unknown and gene deletions ofNET4did not result in a global growth phenotype. Here we show that NET4 functions redundantly with NET3, interacting with RABG3-GTPases at the vacuole to allow for homotypic fusion or, alternatively, the generation of endoplasmic reticulum (ER) - vacuole contact sites. We found that ER-resident NET3 is able to interact with RABG3 residing at the tonoplast and that NET4 interacts with the contact site protein VAP27-1 at the ER. Generation ofnet3 net4triple mutants by CRSIPR-guided mutagenesis helped us to overcome functional redundancy, resulting in impaired plant growth and development. Our results demonstrate how diversification ofNETgenes led to functional redundancy between different family members to create cellular plasticity of vascular plants. We hypothesize that establishment of a direct ER-vacuole connection enables direct lipid and protein transfer which is especially important in young and fast-growing cells. Availability of lipids would facilitate rapidly expanding vacuoles which are the basis for high cell elongation rates and eventually fast plant growth.

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Membrane contact sites and cytoskeleton‐membrane interactions in autophagy

Cited by 11 publications

References 68 publications

Mitochondrial networks of cumulus cells and oocyte quality

Mitochondrial networks of cumulus cells and oocyte quality

Applying Optical Tweezers with TIRF Microscopy to Quantify Physical Interactions Between Organelles in the Plant Endomembrane System

Networked proteins redundantly interact with VAP27 and RABG3 to regulate membrane tethering at the vacuole and beyond

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