PUBLICATION

A novel cis-regulatory element drives early expression of Nkx3.2 in the gnathostome primary jaw joint

Authors

Leyhr, J., Waldmann, L., Filipek-Górniok, B., Zhang, H., Allalou, A., Haitina, T.

ID

ZDB-PUB-221116-4

Date

2022

Source

eLIFE 11: (Journal)

Registered Authors

Filipek-Górniok, Beata, Haitina, Tatjana, Leyhr, Jake, Waldmann, Laura

Keywords

cis-regulatory element, developmental biology, enhancer deletion, evolutionary biology, gnathostome, jaw joint, nkx3.2, optical projection tomography, zebrafish

MeSH Terms

Regulatory Sequences, Nucleic Acid
Zebrafish*/genetics
Jaw
Gene Expression Regulation, Developmental/genetics
Biological Evolution
Gene Deletion
Animals
Vertebrates/genetics
Vertebrates/growth & development
Genome
Lampreys
Mammals/genetics

(all 12)

PubMed

36377467 Full text @ Elife

Abstract

The acquisition of movable jaws was a major event during vertebrate evolution. The role of NK3 homeobox 2 (Nkx3.2) transcription factor in patterning the primary jaw joint of gnathostomes (jawed vertebrates) is well known, however knowledge about its regulatory mechanism is lacking. In this study, we report a proximal enhancer element of Nkx3.2 that is deeply conserved in most gnathostomes but undetectable in the jawless hagfish and lamprey. This enhancer is active in the developing jaw joint region of the zebrafish Danio rerio, and was thus designated as jaw joint regulatory sequence 1 (JRS1). We further show that JRS1 enhancer sequences from a range of gnathostome species, including a chondrichthyan and mammals, have the same activity in the jaw joint as the native zebrafish enhancer, indicating a high degree of functional conservation despite the divergence of cartilaginous and bony fish lineages or the transition of the primary jaw joint into the middle ear of mammals. Finally, we show that deletion of JRS1 from the zebrafish genome using CRISPR/Cas9 results in a significant reduction of early gene expression of nkx3.2 and leads to a transient jaw joint deformation and partial fusion. Emergence of this Nkx3.2 enhancer in early gnathostomes may have contributed to the origin and shaping of the articulating surfaces of vertebrate jaws.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Protruding-mouth	Meckel's cartilage posterior margin fused with palatoquadrate cartilage anterior margin, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Protruding-mouth	Meckel's cartilage-palatoquadrate cartilage joint embryonic skeletal joint morphogenesis decreased process quality, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Protruding-mouth	Meckel's cartilage-palatoquadrate cartilage joint malformed, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Day 5	Meckel's cartilage posterior margin fused with palatoquadrate cartilage anterior margin, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Day 5	Meckel's cartilage-palatoquadrate cartilage joint embryonic skeletal joint morphogenesis decreased process quality, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Day 5	Meckel's cartilage-palatoquadrate cartilage joint malformed, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Days 14-20	Meckel's cartilage posterior margin fused with palatoquadrate cartilage anterior margin, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Days 14-20	Meckel's cartilage-palatoquadrate cartilage joint embryonic skeletal joint morphogenesis decreased process quality, abnormal	Figure 4.
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg	standard conditions	Days 14-20	Meckel's cartilage-palatoquadrate cartilage joint malformed, abnormal	Figure 4.
nkx3-2^uu3731/+	standard conditions	Day 5	Meckel's cartilage-palatoquadrate cartilage joint morphology, normal	Figure 6

1 - 10 of 24

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ba2Tg	Tg(-4.9sox10:EGFP)	Transgenic Insertion
uu2803		Small Deletion	nkx3-2
uu3731		Small Deletion	nkx3-2
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion
zf3982Tg	Tg(rr.jrs1:mCherry)	Transgenic Insertion

1 - 5 of 5

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
rr.jrs1	CRISPR1-rr.jrs1	CRISPR
rr.jrs1	CRISPR2-rr.jrs1	CRISPR

Fish

Fish
ba2Tg; zf3982Tg
nkx3-2^{uu2803/uu2803}; ba2Tg; zf3982Tg
nkx3-2^uu3731/+
nkx3-2^{uu3731/uu3731}
WT
y1Tg; zf3982Tg

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
mCherry	EFG	mCherry

Mapping

Leyhr et al., 2022 ZDB-PUB-221116-4 PMID:36377467

A novel cis-regulatory element drives early expression of Nkx3.2 in the gnathostome primary jaw joint

Leyhr et al., 2022

ZDB-PUB-221116-4
PMID:36377467