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Dryad

RNA seq data set of chick, quail, and duck mandibular primordia at embryonic stage (HH) 37

Cite this dataset

Smith, Spenser S; Chu, Daniel; Schneider, Richard A (2022). RNA seq data set of chick, quail, and duck mandibular primordia at embryonic stage (HH) 37 [Dataset]. Dryad. https://doi.org/10.7272/Q62805W5

Abstract

Jaws are among the most highly adapted and modified structures of vertebrates, and they facilitate complex behaviors like feeding, respiration, and vocalization. For this reason, precise developmental regulation of jaw length is crucial for survival. By comparing jaw development between white Pekin duck (Anas platyrhynchos domestica), Japanese quail (Coturnix coturnix japonica), and chick (Gallus gallus) we have shown in prior work that neural crest mesenchyme (NCM), which is the embryonic progenitor population that gives rise to the jaw skeleton, employs a variety of stage- and species-specific mechanisms to govern jaw length. Duck have much longer jaws compared to those of quail, and during the early migration of NCM from the anterior neural tube, duck embryos allocate more progenitors to the presumptive jaw region. Once these NCM populations arrive, their growth trajectories further diverge due to autonomous molecular programs for proliferation and differentiation that are tied to intrinsic rates of maturation and species-specific regulation of multiple-signaling pathways.

To analyze the species-specific differential expression and regulation of the signaling pathways, which direct jaw morphology, we performed an RNA-seq experiment on chick, quail, and duck mandibular primordia at stage (HH) 37. To increase the spatial resolution the lower jaws were dissected into proximal and distal portions. Oxford Nanopore Technology sequencing was used to preserve the full-length mRNA sequence so that spatial and species-specific differences in the distribution of isoforms could be assayed.

Methods

Lower jaw primordia were dissected from HH37 chick, quail, and duck. The lower jaw primordia were cut in half along the distal-proximal axis and RNA was extracted using RNeasy plus kit. cDNA from HH37 chick, quail, and duck lower jaws was synthesized using an NEB Next Single Cell/Low Input cDNA Synthesis Amplification Module (E6421, New England BioLabs, Ipswich, MA) following PacBio Iso-Seq template preparation guidelines. cDNA was barcoded using the PCR barcoding expansion 1-96 kit (EXP-PBC096, Oxford Nanopore Technologies, Oxford, UK). cDNA samples were pooled, and a sequencing library was prepared using the ligation sequencing kit (SQK-LSK110, Oxford Nanopore Technologies, Oxford, UK). 50 fmol of the final library was loaded onto each of the two PromethION flowcells (v R9.4.1) and the run was performed for 72 hours. Basecalling and de-multiplexing were performed live on the PromethION compute module (Basecaller Version ont-Guppy-for-minKNOW 4.0.11).

Usage notes

Data are sequencing results from long read Oxford Nanopore PromethION RNA seq of the distal and proximal portions of mandibular primordia at embryonic stage (HH) 37. Data are in FASTQ file format that have been Gzipped. If necessary, 7-Zip can be used to unzip the files. Software that can read FASTQ files should be able to open this dataset.

Funding

National Institute of Dental and Craniofacial Research, Award: R01 DE016402

National Institute of Dental and Craniofacial Research, Award: R01 DE025668

National Institute of Dental and Craniofacial Research, Award: F31 DE027283