Evolutionarily conserved aspects of animal nutrient uptake and transport in sea anemone vitellogenesis

Lebouvier, Marion; Miramón-Puértolas, Paula; Steinmetz, Patrick R. H.

doi:10.1016/j.cub.2022.08.039

Cited by 9 publications

(11 citation statements)

References 90 publications

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“…A previously published protocol for the generation of CRISPR/Cas9-mediated knock-in lines in Nematostella (Lebouvier et al, 2022) was used and adapted as follows. Two guide RNA (gRNA) target regions with putative cutting sites located 174bp upstream or 229bp downstream of the Start codon of the Nematostella piwi1 (v1g79423) were designed using CRISPOR (Concordet and Haeussler, 2018).…”

Section: Methodsmentioning

confidence: 99%

“…The urea-based in situ hybridization protocol was adapted from (Sinigaglia et al, 2018), with a series of changes as previously described (Lebouvier et al, 2022). After progressive rehydration in MeOH/PTx (0.3% Triton X-100 in 1xPBS pH 7.4), the tissue was digested with Proteinase K 2.5μg/ml for 5 minutes at room temperature.…”

Section: Colorimetric In Situ Hybridizationmentioning

confidence: 99%

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An adult stem-like cell population generates germline and neurons in the sea anemoneNematostella vectensis

Miramón-Puértolas

Steinmetz

2023

Preprint

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Most genetic research animals (e.g., vertebrates, insects, nematodes) segregate germline and soma during early embryogenesis. In contrast, some highly regenerative bilaterian (e.g., planarians) and non-bilaterian animals (e.g., hydrozoan cnidarians) retain adult stem cells with both germinal and somatic potentials. As these cells have been studied in only few phyla, their biology and evolution remain mostly enigmatic. Here, we aimed to identify and characterize adult stem cells and their cell lineages in the sea anemoneNematostella vectensisby combining gene expression analysis, immunostainings, and meganuclease-mediated and CRISPR/Cas9-mediated knock-in reporter lines of conserved germline and multipotency genes (e.g.,vasa2, piwi1). We found a small population ofvasa2+/piwi1+cells in the gastrodermal folds of juvenile and adult sea anemones that generates germline and a diversity of somatic, mostly proliferative cells. Using a combination ofsoxB(2)+neural progenitor and piwi1 reporter lines, we found that the somatic progeny fromvasa2+/piwi1+cells includessoxB(2)+neural progenitors. Our results strongly support the existence of an adult Vasa2+/Piwi1+ multipotent stem-like cell population that derives both germline and somatic lineages inNematostella. The similarities of lineages and gene expression profiles betweenNematostellaVasa2+/Piwi1+ stem-like cells and hydrozoan interstitial stem cells support their evolutionary conservation among cnidarians.

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Section: Methodsmentioning

confidence: 99%

Section: Colorimetric In Situ Hybridizationmentioning

confidence: 99%

An adult stem-like cell population generates germline and neurons in the sea anemoneNematostella vectensis

Miramón-Puértolas

Steinmetz

2023

Preprint

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“…The mesoglea of cnidarians is composed of a largely acellular, mucoid matrix of collagen containing some cells such as amoebocytes 61 . The mesoglea provides rigidity upon which the muscles of the polyp can act but also facilitates the transport of nutrients 62,63 . Some compounds diffuse through the mesoglea like glucose and fatty acids 62,63 while amoebocytes can also shuttle compounds such as amino acids 62 .…”

Section: Residence In the Mesogleamentioning

confidence: 99%

“…The mesoglea provides rigidity upon which the muscles of the polyp can act but also facilitates the transport of nutrients 62,63 . Some compounds diffuse through the mesoglea like glucose and fatty acids 62,63 while amoebocytes can also shuttle compounds such as amino acids 62 . Ca.…”

Section: Residence In the Mesogleamentioning

confidence: 99%

Discovery of deep-sea coral symbionts from a novel family of marine bacteria, Oceanoplasmataceae, with severely reduced genomes

Vohsen

Gruber‐Vodicka

Meyer

et al. 2022

Preprint

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In addition to abundant animal communities, corals from all ocean depths support diverse microbial associates that are important to coral health. While some of these microbes have been classified taxonomically, understanding the metabolic potential of coral-associated bacteria and how they interact with their coral hosts is limited by a lack of genomic data. One example is Mycoplasma and other members of the class Mollicutes which are widespread coral associates. Here we investigated the association between two novel members of the class Mollicutes and the deep-sea octocoral Callogorgia delta. We screened C. delta, a closely related species C. americana, sediment, and water for mollicutes using 16S metabarcoding. One ASV was found in most colonies screened (99/108) and often dominated the microbiome (up to 99%). Another ASV was detected at lower abundance and prevalence in these corals. Both were absent in all water and were absent or rare in the sediment. We sequenced metagenomes and metatranscriptomes to assemble and annotate genomes and propose the names Ca. Oceanoplasma callogorgiae and Ca. Thalassoplasma callogorgiae. The genomes were small, revealed a reliance on the arginine dihydrolase pathway for ATP production, and contained CRISPR-Cas systems with extensive arrays. CARD-FISH microscopy unveiled an abundant bacterium in the mesoglea which is likely to be Ca. O. callogorgiae. These novel mollicutes cluster with others from diverse invertebrate hosts. Altogether, this work describes the association of these novel mollicutes in C. delta, provides insight into widespread coral associates, and identifies a novel clade of marine mollicutes whose diversity remains largely undiscovered.

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“…To overcome these limitations, endogenous protein tagging with fluorescent markers is a valuable method to recapitulate physiological gene expression and protein localization in vivo. Using CRISPR/Cas9 ( 5 ), homology-directed repair (HDR)–based methods for generating knock-in (KI) lines have been described in two cnidarian species, Nematostella and Hydractinia ( 4 , 6 – 9 ) . However, only a few KI cnidarian lines have been reported, reflecting the current difficulties of achieving protein tagging at the endogenous locus.…”

mentioning

confidence: 99%

Endogenous tagging of multiple cellular components in the sea anemone Nematostella vectensis

Paix

Basu

Steenbergen

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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The cnidarian Nematostella vectensis has developed into a powerful model system to study the mechanisms underlying animal development, regeneration, and evolution. However, despite the significant progress in the molecular and genetic approaches in this sea anemone, endogenous protein tagging is still challenging. Here, we report a robust method for knock in for Nematostella using CRISPR/Cas9. As an outcome, we generate endogenously tagged proteins that label core molecular components of several cellular apparatus, including the nuclear envelope, cytoskeleton, cell adhesion, endoplasmic reticulum, cell trafficking, and extracellular matrix. Using live imaging, we monitor the dynamics of vesicular trafficking and endoplasmic reticulum in embryos, as well as cell contractility during the peristaltic wave of a primary polyp. This advancement in gene editing expands the molecular tool kit of Nematostella and enables experimental avenues to interrogate the cell biology of cnidarians.

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Evolutionarily conserved aspects of animal nutrient uptake and transport in sea anemone vitellogenesis

Cited by 9 publications

References 90 publications

An adult stem-like cell population generates germline and neurons in the sea anemoneNematostella vectensis

An adult stem-like cell population generates germline and neurons in the sea anemoneNematostella vectensis

Discovery of deep-sea coral symbionts from a novel family of marine bacteria, Oceanoplasmataceae, with severely reduced genomes

Endogenous tagging of multiple cellular components in the sea anemone Nematostella vectensis

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