Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development

Icha, Jaroslav; Schmied, Christopher; Sidhaye, Jaydeep; Tomančák, Pavel; Preibisch, Stephan; Norden, Caren

doi:10.3791/53966

Cited by 46 publications

(46 citation statements)

References 49 publications

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“…The imaging setup is described previously 31 . Briefly, single 5-6 days post fertilization (dpf) elavl3 : H2B-GCaMP6f 16 or elavl3 : GCaMP6s 15 larva with the opercula removed 1 day prior was paralyzed with a drop of 1mg/ml alpha-bungarotoxin and then embedded in the capillary with 1% low-melting agarose.…”

Section: Methodsmentioning

confidence: 99%

Neural pathways linking hypoxia with pectoral fin movements in Danio rerio

Yeh

Pao

Villa-Real

et al. 2019

Preprint

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15In most animals, respiratory activity inversely correlates with environmental oxygen 16 levels 1,2 . However, less is known about how the underlying neural circuitry encodes 17 oxygen information and modifies behaviours. Here, we characterize the oxygen sensing 18 circuit and reveal sensory coding principles in a Danio rerio larva, an optically 19 accessible vertebrate that increases respiration and startle-related responses under 20 hypoxia 3 . We observe that cranial sensory neurons receive input from multiple oxygen-21 sensing neuroendocrine cells, and then relay this information to hindbrain targets. 22Moreover, hypoxia evoked increase in cranial sensory dendrite calcium events indicates 23 an oxygen-driven change in input dimensionality, which is also represented in their 24 cytoplasm. Additionally, we estimate that a neural code using cytoplasmic calcium 25 events requires most of the cranial sensory neurons, whereas one integrating input 26 dimensionality needs only a third. Furthermore, we show that purinergic signalling at 27 the neuroendocrine cell-sensory neuron synapses drives hypoxia-induced respiratory 28 changes, independent of serotonin, which triggers startle-related responses. Collectively 29 we demonstrate that oxygen coding employs a "many-to-one" sensory circuit that 30 transforms ambient oxygen into neuronal activity and input dimensionality changes to 31 impact behaviour. More broadly, we suggest that multi-dimensional coding might be a 32 common feature of many-to-one circuit motifs, revealing a function for related circuits 33 across species. 34 35Mammals have evolved specialized neural circuitry to detect and respond to oxygen 36 changes. Neuroendocrine glomus cells in the carotid body sense blood oxygen levels and 37 communicate this information to the carotid sinus nerve, a branch of the glossopharyngeal 38 nerve, and the brainstem 4 . Typically, hypoxia is thought to depolarize glomus cells, which in 39 turn release ATP and acetylcholine 5 activating the carotid sinus nerve resulting in higher 40 respiratory rates 6,7 . In parallel, hypoxia also acts via serotonin and brainstem respiratory 41 circuits to promote arousal 8,9 . While neural circuitry and neurotransmitters driving hypoxia-42 induced responses have been described, how the cranial sensory neurons encode oxygen 43 levels and initiate changes in respiratory behaviours. To gain insights into the neuronal 44 dynamics, we probed these pathways in the transparent zebrafish larva. Teleost fish use both 45 the glossopharyngeal and vagal cranial sensory neurons to sense hypoxia 10 . In zebrafish, 46 these neurons innervate the gill-localized neuroendocrine cells (NECs), potential oxygen 47

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Section: Methodsmentioning

confidence: 99%

Neural pathways linking hypoxia with pectoral fin movements in Danio rerio

Yeh

Pao

Villa-Real

et al. 2019

Preprint

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“…Clearing protocols can ameliorate this issue, but their toxicity is not suitable for live imaging [25]. Moreover, most SPIM publications focus on clear samples, such as zebrafish, which do not encounter such issues during imaging [26,27]. …”

Section: Image Process and Registrationmentioning

confidence: 99%

Comparison and combination of rotational imaging optical coherence tomography and selective plane illumination microscopy for embryonic study

Ran

et al. 2017

Biomed. Opt. Express

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Several optical imaging techniques have been applied for high-resolution embryonic imaging using different contrast mechanisms, each with their own benefits and limitations. In this study, we imaged the same E9.5 mouse embryo with rotational imaging optical coherence tomography (RI-OCT) and selective plane illumination microscopy (SPIM). RI-OCT overcomes optical penetration limits of traditional OCT imaging that prohibit full-body imaging of mouse embryos at later stages by imaging the samples from multiple angles. SPIM enables high-resolution, 3D imaging with less phototoxicity and photobleaching than laser scanning confocal microscopy (LSCM) by illuminating the sample with a focused sheet of light. Side by side comparisons are supplemented with co-registered images. The results demonstrate that SPIM and RI-OCT are highly complementary and could provide more comprehensive tissue characterization for mouse embryonic research. Feneley, and R. P. Harvey, "Cardiac septal and valvular dysmorphogenesis in mice heterozygous for mutations in the homeobox gene Nkx2-5," Circ. Res.

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“…In LSFM, a sample is excited with a thin sheet of laser light and the generated single optical section is captured by a perpendicularly oriented detection lens. This arrangement results in low phototoxicity 22 , because only the plane of the specimen that is imaged gets illuminated. 3-dimensional data (3D) are obtained by moving the sample through the light sheet.…”

Section: Introductionmentioning

confidence: 99%

Imaging plant germline differentiation within Arabidopsis flower by light sheet microscopy

Valuchova

Mikulková

Pecinkova

et al. 2019

Preprint

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In higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

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Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development

Cited by 46 publications

References 49 publications

Neural pathways linking hypoxia with pectoral fin movements in Danio rerio

Neural pathways linking hypoxia with pectoral fin movements in Danio rerio

Comparison and combination of rotational imaging optical coherence tomography and selective plane illumination microscopy for embryonic study

Imaging plant germline differentiation within Arabidopsis flower by light sheet microscopy

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