Microscopy of membrane lipids: how precisely can we define their distribution?

Takatori, Sho; Fujimoto, Toyoshi

doi:10.1042/bse0570081

Cited by 6 publications

(2 citation statements)

References 33 publications

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“…This is particularly evident when attempting to compare staining intensity in brain areas of different fish that are not always perfectly matched. When considered in conjunction with the potential for artefactual changes caused by mechanical tissue manipulation (Griffiths et al 1993 ; Takatori and Fujimoto 2015 ), it is clear that a different approach may be required. These difficulties may explain the relative absence of fluorescent lipid measurements in LSD zebrafish models.…”

Section: Resultsmentioning

confidence: 99%

Visualisation of cholesterol and ganglioside GM1 in zebrafish models of Niemann–Pick type C disease and Smith–Lemli–Opitz syndrome using light sheet microscopy

Cook

Bladen

Smith

et al. 2020

Histochem Cell Biol

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Lysosomal storage diseases are the most common cause of neurodegeneration in children. They are characterised at the cellular level by the accumulation of storage material within lysosomes. There are very limited therapeutic options, and the search for novel therapies has been hampered as few good small animal models are available. Here, we describe the use of light sheet microscopy to assess lipid storage in drug and morpholino induced zebrafish models of two diseases of cholesterol homeostasis with lysosomal dysfunction: First, Niemann–Pick type C disease (NPC), caused by mutations in the lysosomal transmembrane protein NPC1, characterised by intralysosomal accumulation of cholesterol and several other lipids. Second, Smith–Lemli–Opitz syndrome (SLOS), caused by mutations in 7-dehydrocholesterol reductase, which catalyses the last step of cholesterol biosynthesis and is characterised by intralysosomal accumulation of dietary cholesterol. This is the first description of a zebrafish SLOS model. We find that zebrafish accurately model lysosomal storage and disease-specific phenotypes in both diseases. Increased cholesterol and ganglioside GM1 were observed in sections taken from NPC model fish, and decreased cholesterol in SLOS model fish, but these are of limited value as resolution is poor, and accurate anatomical comparisons difficult. Using light sheet microscopy, we were able to observe lipid changes in much greater detail and identified an unexpected accumulation of ganglioside GM1 in SLOS model fish. Our data demonstrate, for the first time in zebrafish, the immense potential that light sheet microscopy has in aiding the resolution of studies involving lysosomal and lipid disorders.

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Section: Resultsmentioning

confidence: 99%

Visualisation of cholesterol and ganglioside GM1 in zebrafish models of Niemann–Pick type C disease and Smith–Lemli–Opitz syndrome using light sheet microscopy

Cook

Bladen

Smith

et al. 2020

Histochem Cell Biol

View full text Add to dashboard Cite

show abstract

“…Already in the 1960s, Cope and Williams quantified extraction of various lipids species after aldehyde fixation, comparing different organic solvents and resins. Post-fixation with osmium tetroxide before dehydration reduced the extraction of lipids to different degrees, depending on the lipids species and on the resin used (reviewed in refs [15,16]). Extraction was reduced for some lipids species (e.g.…”

Section: Sample Preservation and Consequences For Membranesmentioning

confidence: 99%

The sleeping beauty kissed awake: new methods in electron microscopy to study cellular membranes

Chlanda

Locker

2017

Biochemical Journal

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Electron microscopy (EM) for biological samples, developed in the 1940-1950s, changed our conception about the architecture of eukaryotic cells. It was followed by a period where EM applied to cell biology had seemingly fallen asleep, even though new methods with important implications for modern EM were developed. Among these was the discovery that samples can be preserved by chemical fixation and most importantly by rapid freezing without the formation of crystalline ice, giving birth to the world of cryo-EM. The past 15-20 years are hallmarked by a tremendous interest in EM, driven by important technological advances. Cryo-EM, in particular, is now capable of revealing structures of proteins at a near-atomic resolution owing to improved sample preparation methods, microscopes and cameras. In this review, we focus on the challenges associated with the imaging of membranes by EM and give examples from the field of host-pathogen interactions, in particular of virus-infected cells. Despite the advantages of imaging membranes under native conditions in cryo-EM, conventional EM will remain an important complementary method, in particular if large volumes need to be imaged.

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Detectors for evaluating the cellular landscape of sphingomyelin- and cholesterol-rich membrane domains

Kishimoto

Ishitsuka

Kobayashi

2016

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Microscopy of membrane lipids: how precisely can we define their distribution?

Cited by 6 publications

References 33 publications

Visualisation of cholesterol and ganglioside GM1 in zebrafish models of Niemann–Pick type C disease and Smith–Lemli–Opitz syndrome using light sheet microscopy

Visualisation of cholesterol and ganglioside GM1 in zebrafish models of Niemann–Pick type C disease and Smith–Lemli–Opitz syndrome using light sheet microscopy

The sleeping beauty kissed awake: new methods in electron microscopy to study cellular membranes

Detectors for evaluating the cellular landscape of sphingomyelin- and cholesterol-rich membrane domains

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