A Cryptochrome adopts distinct moon- and sunlight states and functions as sun- versus moonlight interpreter in monthly oscillator entrainment

Poehn, Birgit; Krishnan, Shruthi; Zurl, Martin; Ćorić, Aida; Rokvic, Dunja; Häfker, N. Sören; Jaenicke, Elmar; Arboleda, Enrique; Orel, Lukas; Raible, Florian; Wolf, Eva; Tessmar-Raible, Kristin

doi:10.1038/s41467-022-32562-z

Cited by 25 publications

(51 citation statements)

References 51 publications

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“…One of the interesting aspects about Platynereis dumerilii is its use as a functional molecular model system for chronobiological analyses (Özpolat et al, 2021b; Raible and Tessmar-Raible, 2014). At least two endogenous timing systems – a ~24hr (plastic circadian-circalunidian) and a monthly (circalunar) one – co-exist in Platynereis and are influenced by ambient light conditions (Zantke et al, 2013) (Poehn et al, 2022; Zurl et al, 2022). Period is a key gene of the circadian clock (Glossop and Hardin, 2002).…”

Section: Resultsmentioning

confidence: 99%

“…To test the compatibility of SHInE with IHC, immunostainings were performed on the same samples using antibodies raised against Platynereis L-Cryptochrome (L-Cry). L-Cry is a photoreceptor that shares regions of expression with period and is well described also on the sub-cellular level for P.dumerilii (Poehn et al, 2022; Zurl et al, 2022). In line with the published expression patterns, the acquisition of SHInE-processed heads revealed expression of L-Cry in the medial brain nuclei and in the eyes (Figure 2C,D), in the same region as period RNA (Figure 2A,D).…”

Section: Resultsmentioning

confidence: 99%

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A fast and versatile method for simultaneous HCR, immunohistochemistry and EdU labeling (SHInE)

Ćorić

Stockinger

Schaffer

et al. 2022

Preprint

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Access to newer, fast and cheap sequencing techniques, particularly on the single-cell level, have made transcriptomic data of tissues or single cells accessible to many researchers. As a consequence, there is increased need for in situ visualization of gene expression or encoded proteins to validate, localize or help interpret such sequencing data, as well as put them in context with cellular proliferation. A particular challenge for labeling and imaging transcripts are complex tissues that are often opaque and/or pigmented, preventing easy visual inspection. Here we introduce a versatile protocol that combines in situ hybridization chain reaction (HCR), immunohistochemistry (IHC) and proliferative cell labeling using 5-ethynyl-2′-deoxyuridine (EdU), and demonstrate its compatibility with tissue clearing. As a proof-of-concept, we show that our protocol allows for the parallel analysis of cell proliferation, gene expression and protein localization in bristleworm heads and trunks.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

A fast and versatile method for simultaneous HCR, immunohistochemistry and EdU labeling (SHInE)

Ćorić

Stockinger

Schaffer

et al. 2022

Preprint

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“…This culturing technique may not be suitable for all experimental purposes. For instance, some teams study biological rhythms, especially those that are controlled by the lunar cycle (Poehn et al, 2022).…”

Section: Suitability Of the Present Culture Methods For Different Typ...mentioning

confidence: 99%

Fast Cycling Culture of the Marine AnnelidPlatynereis dumerilii

Legras

Ghisleni

Soilihi

et al. 2023

Preprint

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Platynereis dumerilii, a marine annelid, is a model animal that has gained popularity in various fields such as developmental biology, biological rhythms, nervous system organization and physiology, behaviour, reproductive biology, and epigenetic regulation. The transparency of P. dumerilii tissues at all developmental stages makes it easy to perform live microscopic imaging of all cell types. In addition, the slow-evolving genome of P. dumerilii and its phylogenetic position as a representative of the vast branch of Lophotrochozoans add to its evolutionary significance. Although P. dumerilii is amenable to transgenesis and CRISPR-Cas9 knockouts, its relatively long and indefinite life cycle, as well as its semelparous reproduction have been hindrances to its adoption as a reverse genetics model. To overcome this limitation, an adapted culturing method has been developed allowing much faster life cycling, with median reproductive age at 15 weeks instead of 6-8 months using the traditional protocol. A low worm density in boxes and a strictly controlled feeding regime are important factors for the rapid growth and health of the worms. Moreover, a genetic selection for fast-reproducing individuals has been applied to isolate a "Fast Forward" strain that can be used for egg microinjection. This culture method has several advantages, such as being much more compact, not requiring air bubbling or an artificial moonlight regime for synchronized sexual maturation, and necessitating only limited water change. A full protocol for worm care and handling is provided.

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“…Oligomerisation of proteins can change their structural stability, activity, and mechanisms of action 1,2 and is important in numerous processes in living organisms. Examples from the cryptochrome (Cry) protein family include a tetrameric structure of Arabidopsis thaliana Cry2 (AtCry2) important in photoactivation 3 , a proposed dimer of L-Cry involved in the reproduction cycle of the bristle worm, Platynereis dumerilii 4 , and dimerisation of AtCry1 5,6 .…”

Section: Introductionmentioning

confidence: 99%

Dimerisation of European robin cryptochrome 4a

Hanić

Antill

Gehrckens

et al. 2023

Preprint

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Homo-dimer formation is important for the function of many proteins. Although dimeric forms of cryptochromes (Cry) have been found by crystallography and were recently observed in vitro for European robin Cry4a, little is known about the dimerisation of avian cryptochromes and the role it could play in the mechanism of magnetic sensing in migratory birds. Here we present a combined experimental and computational investigation of the dimerisation of robin Cry4a resulting from covalent and non-covalent interactions. Experimental studies using native mass spectrometry, mass spectrometric analysis of disulphide bonds, chemical cross-linking and photometric measurements show that disulphide-linked dimers are routinely formed, the most likely cysteines being C317 and C412. Computational modelling and molecular dynamics simulations were used to generate and assess a number of possible dimer structures. The relevance of these findings to the proposed role of Cry4a in avian magnetoreception is discussed.

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A Cryptochrome adopts distinct moon- and sunlight states and functions as sun- versus moonlight interpreter in monthly oscillator entrainment

Cited by 25 publications

References 51 publications

A fast and versatile method for simultaneous HCR, immunohistochemistry and EdU labeling (SHInE)

A fast and versatile method for simultaneous HCR, immunohistochemistry and EdU labeling (SHInE)

Fast Cycling Culture of the Marine AnnelidPlatynereis dumerilii

Dimerisation of European robin cryptochrome 4a

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