Whole mount in situ hybridization techniques for analysis of the spatial distribution of mRNAs in sea urchin embryos and early larvae

Erkenbrack, Eric M.; Croce, Jenifer C.; Miranda, Esther; Gautam, Sujan; Martínez-Bartolomé, Marina; Yaguchi, Shunsuke; Range, Ryan

doi:10.1016/bs.mcb.2019.01.003

Cited by 8 publications

(7 citation statements)

References 13 publications

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“…Marine organisms exhibit remarkable variability among different species. The sample preparation method for spatial transcriptomics was optimized based on previous results obtained using fixed and embedded methods for in situ hybridization and single-molecule RNA fluorescence in situ hybridization ( 63 – 65 ). To ensure the reproducibility of the atlas, we combined three replicates of scRNA-seq and six sections of Stereo-seq.…”

Section: Discussionmentioning

confidence: 99%

Spatially resolved single-cell atlas of ascidian endostyle provides insight into the origin of vertebrate pharyngeal organs

Jiang,

Han,

Wei

et al. 2024

Sci. Adv.

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The pharyngeal endoderm, an innovation of deuterostome ancestors, contributes to pharyngeal development by influencing the patterning and differentiation of pharyngeal structures in vertebrates; however, the evolutionary origin of the pharyngeal organs in vertebrates is largely unknown. The endostyle, a distinct pharyngeal organ exclusively present in basal chordates, represents a good model for understanding pharyngeal organ origins. Using Stereo-seq and single-cell RNA sequencing, we constructed aspatially resolved single-cell atlas for the endostyle of the ascidian Styela clava . We determined the cell composition of the hemolymphoid region, which illuminates a mixed ancestral structure for the blood and lymphoid system. In addition, we discovered a cluster of hair cell–like cells in zone 3, which has transcriptomic similarity with the hair cells of the vertebrate acoustico-lateralis system. These findings reshape our understanding of the pharynx of the basal chordate and provide insights into the evolutionary origin of multiplexed pharyngeal organs.

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Section: Discussionmentioning

confidence: 99%

Spatially resolved single-cell atlas of ascidian endostyle provides insight into the origin of vertebrate pharyngeal organs

Jiang,

Han,

Wei

et al. 2024

Sci. Adv.

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“…Alkaline phosphatase and two-color fluorescent in situ hybridization were carried out as previously described. 21 , 57 , 104 Embryos were fixed in 4% paraformaldehyde in ASW for 1 hour at room temperature. Probes, labeled with Digoxigenin, were hybridized for 5-7 days.…”

Section: Methodsmentioning

confidence: 99%

Evolutionarily conserved Wnt/Sp5 signaling is critical for anterior-posterior axis patterning in sea urchin embryos

Gautam,

Fenner,

Wang

et al. 2024

iScience

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“…The samples were incubated with RNA probes for foxQ2 (HPU_15608), rx (HPU_04689), hbn (HPU_04688), lefty (HPU_15030), neurogenin (HPU_07864), choline acetyltransferase (HPU_01496), msi1 (HPU_03784) and tryptophan 5-hydroxylase ( tph ; HPU_21307) ( Ayala et al, 2007 ) at a final concentration of 0.4–1.2 ng/μL at 50°C for 5 days. The probes were detected with the Tyramide Signal Amplification Plus System (TSA; Akoya Biosciences, Marlborough, MA, United States) or Alkaline phosphatase-based chromogenic system described previously ( Erkenbrack et al, 2019 ).…”

Section: Methodsmentioning

confidence: 99%

Rx and its downstream factor, Musashi1, is required for establishment of the apical organ in sea urchin larvae

Yaguchi,

Yaguchi

2023

Front. Cell Dev. Biol.

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Acetylcholine, a vital neurotransmitter, plays a multifarious role in the brain and peripheral nervous system of various organisms. Previous research has demonstrated the proximity of cholinergic neurons to serotonergic neurons in the apical organ of sea urchin embryos. While several transcription factors have been identified as playing a role in the development of serotonergic neurons in this region of a sea urchin, Hemicentrotus pulcherrimus, comparatively little is known about the specific transcription factors and their spatiotemporal expression patterns that regulate the development of cholinergic neurons. In this study, we establish the requirement of the transcription factor Rx for the development of cholinergic neurons in the apical organ of the species. Furthermore, we investigate the role of the RNA-binding protein Musashi1, known to be involved in neurogenesis, including cholinergic neurons in other organisms, and demonstrate that it is a downstream factor of Rx, and that choline acetyltransferase expression is suppressed in Musashi1 downregulated embryos. Our research also highlights the intricate network formed by neurons and other cells in and around the apical organ of sea urchin larvae through axons and dendrites, providing possibility for a systematic and complexed neural pattern like those of the brain in other organisms.

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Whole mount in situ hybridization techniques for analysis of the spatial distribution of mRNAs in sea urchin embryos and early larvae

Cited by 8 publications

References 13 publications

Spatially resolved single-cell atlas of ascidian endostyle provides insight into the origin of vertebrate pharyngeal organs

Spatially resolved single-cell atlas of ascidian endostyle provides insight into the origin of vertebrate pharyngeal organs

Evolutionarily conserved Wnt/Sp5 signaling is critical for anterior-posterior axis patterning in sea urchin embryos

Rx and its downstream factor, Musashi1, is required for establishment of the apical organ in sea urchin larvae

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