In situ hybridization of chick and mouse embryonic tissue

Sockanathan, Shan

doi:10.1038/protex.2015.040

Cited by 2 publications

(2 citation statements)

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“…A Ventana Discovery XT system (Roche-Ventana Medical Systems) was used to perform in situ hybridization for mouse Acan. In situ hybridization for chick Hoxa2 was performed as previously described (Schaeren-Wiemers and Gerfin-Moser, 1993;Sockanathan, 2015) with slight modifications. Briefly, the sections were fixed in 4% PFA in PBS for 5-10 min followed by 10 min acetylation (21 μM HCl, 100 μM triethanolamine in water, 0.25% acetic anhydride) at RT.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Differing contributions of the first and second pharyngeal arches to tympanic membrane formation in the mouse and chick

et al. 2017

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We have proposed that independent origins of the tympanic membrane (TM), consisting of the external auditory meatus (EAM) and first pharyngeal pouch, are linked with distinctive middle ear structures in terms of dorsal-ventral patterning of the pharyngeal arches during amniote evolution. However, previous studies have suggested that the first pharyngeal arch (PA1) is crucial for TM formation in both mouse and chick. In this study, we compare TM formation along the anterior-posterior axis in these animals using expression as a marker of the second pharyngeal arch (PA2). In chick, the EAM begins to invaginate at the surface ectoderm of PA2, not at the first pharyngeal cleft, and the entire TM forms in PA2. Chick-quail chimera that have lost PA2 and duplicated PA1 suggest that TM formation is achieved by developmental interaction between a portion of the EAM and the columella auris in PA2, and that PA1 also contributes to formation of the remaining part of the EAM. By contrast, in mouse, TM formation is highly associated with an interdependent relationship between the EAM and tympanic ring in PA1.

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Section: In Situ Hybridizationmentioning

confidence: 99%

Differing contributions of the first and second pharyngeal arches to tympanic membrane formation in the mouse and chick

et al. 2017

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“…Probes are listed in (Table S3). In situ experiments in sections were performed according to (Sockanathan and Sockanathan, 2015). Cryostat sections were prepared after fixing embryos in 4% PFA solution at 4ºC overnight.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Developmental and Evolutionary Comparative Analysis of a Regulatory Landscape in Mammals and Birds

Hintermann

Guerreiro

Bolt

et al. 2021

Preprint

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Modifications in gene regulation during development are considered to be a driving force in the evolution of organisms. Part of these changes involve rapidly evolving cis-regulatory elements (CREs), which interact with their target genes through higher-order 3D chromatin structures. How such 3D architectures and variations in CREs contribute to transcriptional evolvability nevertheless remains elusive. During vertebrate evolution, Hox genes were redeployed in different organs in a class-specific manner, while maintaining the same basic function in organizing the primary body axis. Since a large part of the relevant enhancers are located in a conserved regulatory landscape, this gene cluster represents an interesting paradigm to study the emergence of regulatory innovations. In this work, we analyzed Hoxd gene regulation in both murine vibrissae and chicken feather primordia, two mammalian- and avian-specific skin appendages which express different subsets of Hoxd genes, and compared their regulatory modalities with the regulations at work during the elongation of the posterior trunk, a mechanism highly conserved in amniotes. We show that in the former two structures, distinct subsets of Hoxd genes are contacted by different lineage-specific enhancers, likely as a result of using an ancestral chromatin topology as an evolutionary playground, whereas the regulations implemented in the mouse and chicken embryonic trunk rely more on conserved CREs. Nevertheless, a high proportion of these non-coding sequences active in the trunk appear to have functionally diverged between the two species, suggesting that transcriptional robustness is maintained despite a considerable divergence in CRE sequences, an observation supported by a genome-wide comparative approach.

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In situ hybridization of chick and mouse embryonic tissue

Abstract: The detection of transcripts in sectioned tissue by in situ hybridization is a useful method to localize sites of gene expression during development. This protocol is one that we have modified from Scharen-Wiemers and Gerfin-Moser Histochemistry 100:431-440.

Cited by 2 publications

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Differing contributions of the first and second pharyngeal arches to tympanic membrane formation in the mouse and chick

Differing contributions of the first and second pharyngeal arches to tympanic membrane formation in the mouse and chick

Developmental and Evolutionary Comparative Analysis of a Regulatory Landscape in Mammals and Birds

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