X-Ray Microscopy of the Larval Crustacean Brain

Krieger, Jakob; Spitzner, Franziska

doi:10.1007/978-1-4939-9732-9_14

Cited by 7 publications

(10 citation statements)

References 34 publications

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“…Iodine was subsequently washed out of the samples for 3 × 10 min with ethanol, and the samples were dried with an fully automatic critical point dryer Leica EM CPD300. For more details on the μCT‐preparations, see also Sombke, Lipke, Michalik, Uhl, and Harzsch () with slight modifications after Krieger and Spitzner ().…”

Section: Methodsmentioning

confidence: 99%

Masters of communication: The brain of the banded cleaner shrimp Stenopus hispidus (Olivier, 1811) with an emphasis on sensory processing areas

Krieger

Hörnig

Sandeman

et al. 2019

J of Comparative Neurology

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The pan‐tropic cleaner shrimp Stenopus hispidus (Crustacea, Stenopodidea) is famous for its specific cleaning behavior in association with client fish and an exclusively monogamous life‐style. Cleaner shrimps feature a broad communicative repertoire, which is considered to depend on superb motor skills and the underlying mechanosensory circuits in combination with sensory organs. Their most prominent head appendages are the two pairs of very long biramous antennules and antennae, which are used both for attracting client fish and for intraspecific communication. Here, we studied the brain anatomy of several specimens of S. hispidus using histological sections, immunohistochemical labeling as well as X‐ray microtomography in combination with 3D reconstructions. Furthermore, we investigated the morphology of antennules and antennae using fluorescence and scanning electron microscopy. Our analyses show that in addition to the complex organization of the multimodal processing centers, especially chemomechanosensory neuropils associated with the antennule and antenna are markedly pronounced when compared to the other neuropils of the central brain. We suggest that in their brains, three topographic maps are present corresponding to the sensory appendages. The brain areas which provide the neuronal substrate for these maps share distinct structural similarities to a unique extent in decapods, such as size and characteristic striated and perpendicular layering. We discuss our findings with respect to the sensory landscape within animal's habitat. In an evolutionary perspective, the cleaner shrimp's brain is an excellent example of how sensory potential and functional demands shape the architecture of primary chemomechanosensory processing areas.

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Section: Methodsmentioning

confidence: 99%

Masters of communication: The brain of the banded cleaner shrimp Stenopus hispidus (Olivier, 1811) with an emphasis on sensory processing areas

Krieger

Hörnig

Sandeman

et al. 2019

J of Comparative Neurology

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“…Here, we briefly summarise methodological steps of the sample preparation for a workflow enabling a consecutive immunohistochemical and/or histological investigation (for further details of other possible tissue preparations see our extensive manual on µCT in decapod larvae [122]). First of all, the larvae need to be properly fixed.…”

Section: Description Of the Methodsmentioning

confidence: 99%

“…After dehydration and before scanning, the tissues should be incubated in 2% iodine in methanol for contrast-enhancement for at least 24 h at room temperature. After two to three washing steps using pure methanol, the larvae can be directly scanned ideally in sealable plastic chambers filled with fresh methanol (for further details on mounting of samples, see [122]). After successful scanning, the sample needs to be rehydrated in a decreasing series of methanol in TRIS-buffer and finally be washed in several changes of PBS-TX (0.3% Triton X-100 in 0.1 M PBS) to remove any reminiscent iodine from the tissue.…”

Section: Description Of the Methodsmentioning

confidence: 99%

“…What is more, sample preparation and µCT, like most other techniques, also have specific disadvantages. For delivering good contrast, especially soft tissues need to be fixed and incubated in contrastenhancing agents in advance which often cause tissue modifications such as shrinking (see [111,122]). However, by using methanol as outlined above, we obtained excellent results for later immunohistochemical labelling against anti-synapsin (SYNORF1) in combination with histochemical detection of the cell proliferation marker EdU previously in vivo injected (Krieger, unpublished).…”

Section: Additional Commentsmentioning

confidence: 99%

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Methods to study organogenesis in decapod crustacean larvae II: analysing cells and tissues

et al. 2021

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Cells and tissues form the bewildering diversity of crustacean larval organ systems which are necessary for these organisms to autonomously survive in the plankton. For the developmental biologist, decapod crustaceans provide the fascinating opportunity to analyse how the adult organism unfolds from organ Anlagen compressed into a miniature larva in the sub-millimetre range. This publication is the second part of our survey of methods to study organogenesis in decapod crustacean larvae. In a companion paper, we have already described the techniques for culturing larvae in the laboratory and dissecting and chemically fixing their tissues for histological analyses. Here, we review various classical and more modern imaging techniques suitable for analyses of eidonomy, anatomy, and morphogenetic changes within decapod larval development, and protocols including many tips and tricks for successful research are provided. The methods cover reflected-light-based methods, autofluorescence-based imaging, scanning electron microscopy, usage of specific fluorescence markers, classical histology (paraffin, semithin and ultrathin sectioning combined with light and electron microscopy), X-ray microscopy (µCT), immunohistochemistry and usage of in vivo markers. For each method, we report our personal experience and give estimations of the method’s research possibilities, the effort needed, costs and provide an outlook for future directions of research.

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“…Carcinus maenas STH, µCT, 3D, IHC [175][176][177][178] Hyas araneus IHC, PM, STH [179][180][181][182][183] Pachygrapsus marmoratus STH, 3D [184] Porcellana platycheles STH, 3D [184] Cherax destructor PM [171,185,186] Homarus americanus STH, PM, IHC [183,[186][187][188][189][190][191][192] Homarus gammarus STH, TEM, IHC [161,162] Hippolyte inermis STH, 3D [184] should follow standard procedures of sampling theory. We refer to key references for the extensive discussion of experimental design [79][80][81].…”

Section: Structure Of the Larval Cns And Neurogenesismentioning

confidence: 99%

Methods to study organogenesis in decapod crustacean larvae. I. larval rearing, preparation, and fixation

et al. 2021

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Crustacean larvae have served as distinguished models in the field of Ecological Developmental Biology (“EcoDevo”) for many decades, a discipline that examines how developmental mechanisms and their resulting phenotype depend on the environmental context. A contemporary line of research in EcoDevo aims at gaining insights into the immediate tolerance of organisms and their evolutionary potential to adapt to the changing abiotic and biotic environmental conditions created by anthropogenic climate change. Thus, an EcoDevo perspective may be critical to understand and predict the future of organisms in a changing world. Many decapod crustaceans display a complex life cycle that includes pelagic larvae and, in many subgroups, benthic juvenile–adult stages so that a niche shift occurs during the transition from the larval to the juvenile phase. Already at hatching, the larvae possess a wealth of organ systems, many of which also characterise the adult animals, necessary for autonomously surviving and developing in the plankton and suited to respond adaptively to fluctuations of environmental drivers. They also display a rich behavioural repertoire that allows for responses to environmental key factors such as light, hydrostatic pressure, tidal currents, and temperature. Cells, tissues, and organs are at the basis of larval survival, and as the larvae develop, their organs continue to grow in size and complexity. To study organ development, researchers need a suite of state-of-the-art methods adapted to the usually very small size of the larvae. This review and the companion paper set out to provide an overview of methods to study organogenesis in decapod larvae. This first section focuses on larval rearing, preparation, and fixation, whereas the second describes methods to study cells, tissues, and organs.

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X-Ray Microscopy of the Larval Crustacean Brain

Cited by 7 publications

References 34 publications

Masters of communication: The brain of the banded cleaner shrimp Stenopus hispidus (Olivier, 1811) with an emphasis on sensory processing areas

Masters of communication: The brain of the banded cleaner shrimp Stenopus hispidus (Olivier, 1811) with an emphasis on sensory processing areas

Methods to study organogenesis in decapod crustacean larvae II: analysing cells and tissues

Methods to study organogenesis in decapod crustacean larvae. I. larval rearing, preparation, and fixation

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