Serial Block Face‐Scanning Electron Microscopy as a Burgeoning Technology

Marshall, Andrea G.; Neikirk, Kit; Stephens, Dominique C.; Vang, Larry; Vue, Zer; Beasley, Heather K.; Crabtree, Amber; Scudese, Estêvão; Lopez, Edgar Garza; Shao, Bryanna; Krystofiak, Evan; Rutledge, Steve; Davis, John E.; Murray, Sandra A.; Damo, Steven M.; Katti, Prasanna; Hinton, Antentor

doi:10.1002/adbi.202300139

Cited by 18 publications

(11 citation statements)

References 60 publications

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“…These techniques are primarily employed for observing samples at the subcellular level and can also handle observations at the cellular and tissue levels. However, as of now, there are limitations on the size of samples these techniques can observe, typically not exceeding 1 cubic millimetre, 29 with a general requirement of 500 µm × 500 µm. 30 For plant floral organs, their sizes often surpass this limitation.…”

Section: Discussionmentioning

confidence: 99%

The fertility tracks of pollen tube in the ovary of Solanum nigrum by three‐dimensional reconstruction

Ming,

Chen,

et al. 2023

Journal of Microscopy

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In this paper, we present an enhanced method for automatically capturing a large number of consecutive paraffin sections using a microscope. Leveraging these microstructural images, we employed three‐dimensional visualisation and reconstruction techniques to investigate the dispersal growth process of pollen tube bundles upon entering the ovary of Solanum nigrum. Additionally, we explored their behaviour within different ovules and examined the relationship between the germination rate of seeds and the fertilisation process. Our findings reveal that despite the abundance of Solanum nigrum seeds, only a fraction of them is capable of successful germination. The germination rate of seeds is closely related to whether fertilisation of the ovules and pollen tubes is completed. Due to the limited number of pollen tubes entering the ovary, only a portion of the ovules can be fertilised. The proportion of fertilised ovules positively correlates with the germination rate of the seeds. Through three‐dimensional reconstruction, we observed a phenomenon of proximity during the pollination process, wherein ovules closer to the pollen tube bundles are more likely to be fertilised. Furthermore, fertilised ovules exhibited significant changes in morphology and embryo sac structure. The number of fertilised ovules directly impacts the germination rate of wild Solanum nigrum seeds. Although all Solanum nigrum ovules have the potential to develop into seeds, most seeds originating from unfertilised ovules are unable to germinate normally, resulting in an incomplete germination rate of seeds and preventing it from reaching 100%.

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Section: Discussionmentioning

confidence: 99%

The fertility tracks of pollen tube in the ovary of Solanum nigrum by three‐dimensional reconstruction

Ming,

Chen,

et al. 2023

Journal of Microscopy

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“…Relative to WT, Opa1-like KD trended toward an increased MERC contact length (Figure 1z), decreased the mean MERC distance (Figure 1aa), increased the percentage of mitochondrial coverage by ER (Figure 1ab), and increased the percentage of ER coverage by mitochondria (Figure 1ac), all suggesting that Opa1-like deficiency increased MERC tethering. Because TEM only captures structural MERC changes in two dimensions (2D), SBF-SEM, which offers numerous advantages in volumetric renderings (Marshall et al, 2023),…”

Section: Opa1-like Kd In Drosophila Flight Muscle Alters Mitochondria...mentioning

confidence: 99%

ATF4‐dependent increase in mitochondrial‐endoplasmic reticulum tethering following OPA1 deletion in skeletal muscle

Hinton,

Katti,

Mungai

et al. 2024

Journal Cellular Physiology

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Mitochondria and endoplasmic reticulum (ER) contact sites (MERCs) are protein‐ and lipid‐enriched hubs that mediate interorganellar communication by contributing to the dynamic transfer of Ca2+, lipid, and other metabolites between these organelles. Defective MERCs are associated with cellular oxidative stress, neurodegenerative disease, and cardiac and skeletal muscle pathology via mechanisms that are poorly understood. We previously demonstrated that skeletal muscle‐specific knockdown (KD) of the mitochondrial fusion mediator optic atrophy 1 (OPA1) induced ER stress and correlated with an induction of Mitofusin‐2, a known MERC protein. In the present study, we tested the hypothesis that Opa1 downregulation in skeletal muscle cells alters MERC formation by evaluating multiple myocyte systems, including from mice and Drosophila, and in primary myotubes. Our results revealed that OPA1 deficiency induced tighter and more frequent MERCs in concert with a greater abundance of MERC proteins involved in calcium exchange. Additionally, loss of OPA1 increased the expression of activating transcription factor 4 (ATF4), an integrated stress response (ISR) pathway effector. Reducing Atf4 expression prevented the OPA1‐loss‐induced tightening of MERC structures. OPA1 reduction was associated with decreased mitochondrial and sarcoplasmic reticulum, a specialized form of ER, calcium, which was reversed following ATF4 repression. These data suggest that mitochondrial stress, induced by OPA1 deficiency, regulates skeletal muscle MERC formation in an ATF4‐dependent manner.

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“…178 Threedimensional microscopy techniques can provide further detailed examination of individual mitochondria and unique phenotypes, 179 which have not yet been comprehensively studied in HIV. Previously, using serial block face-scanning electron microscopy, 180,181 we elucidated how 3D mitochondria structure changed across aging in murine cardiac, skeletal, and brown adipose tissue, giving far greater insight into mitochondrial morphology (eg, swelling, fragmentation, or cristae remodeling) than traditional fluorescent techniques. 165,166,182 Similarly, focus ion beam scanning electron microscopy 183 (1) Following HIV infection, mitochondria can be affected by Vpr (viral protein R), 192,193 which can form the VprBP-DDB1 (damage-specific DNAbinding protein 1)-CUL4A (cullin 4A) ubiquitin ligase complex to reduce MFN2 (mitofusin 2) expression and DRP1 (dynamin-related protein 1).…”

Section: Mitochondrial Dynamics Beyond Opa1 and Drp1mentioning

confidence: 99%

Connection Between HIV and Mitochondria in Cardiovascular Disease and Implications for Treatments

Hinton,

N’jai,

Vue

et al. 2024

Circulation Research

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HIV infection and antiretroviral therapy alter mitochondrial function, which can progressively lead to mitochondrial damage and accelerated aging. The interaction between persistent HIV reservoirs and mitochondria may provide insight into the relatively high rates of cardiovascular disease and mortality in persons living with HIV. In this review, we explore the intricate relationship between HIV and mitochondrial function, highlighting the potential for novel therapeutic strategies in the context of cardiovascular diseases. We reflect on mitochondrial dynamics, mitochondrial DNA, and mitochondrial antiviral signaling protein in the context of HIV. Furthermore, we summarize how toxicities related to early antiretroviral therapy and current highly active antiretroviral therapy can contribute to mitochondrial dysregulation, chronic inflammation, and poor clinical outcomes. There is a need to understand the mechanisms and develop new targeted therapies. We further consider current and potential future therapies for HIV and their interplay with mitochondria. We reflect on the next-generation antiretroviral therapies and HIV cure due to the direct and indirect effects of HIV persistence, associated comorbidities, coinfections, and the advancement of interdisciplinary research fields. This includes exploring novel and creative approaches to target mitochondria for therapeutic intervention.

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Serial Block Face‐Scanning Electron Microscopy as a Burgeoning Technology

Cited by 18 publications

References 60 publications

The fertility tracks of pollen tube in the ovary of Solanum nigrum by three‐dimensional reconstruction

The fertility tracks of pollen tube in the ovary of Solanum nigrum by three‐dimensional reconstruction

ATF4‐dependent increase in mitochondrial‐endoplasmic reticulum tethering following OPA1 deletion in skeletal muscle

Connection Between HIV and Mitochondria in Cardiovascular Disease and Implications for Treatments

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