Npr3 regulates neural crest and cranial placode progenitors formation through its dual function as clearance and signaling receptor
- Department of Molecular Pathobiology, New York University, College of Dentistry, United States
Figures
Figure 1 with 1 supplement
Npr3, a target of Pax3 and Zic1, is expressed in developing NC and CP.
(A) By ISH, npr3 is detected at the neural plate border at neurula stages (NF stage 13–17), and persists in the NC (yellow arrowheads) and CP (purple arrowheads) forming regions (NF stage 22–26). npr…
Figure 1—figure supplement 1
Developmental RT-PCR of natriuretic peptide signaling pathway components.
Developmental qRT-PCR expression profile of npr1, npr2, npr3, nppa, nppb and nppc in two biological replicates (A and B) of WT Xenopus embryos. NF stages are indicated on x-axis, values are …
Figure 2 with 2 supplements
Npr3 is required for both NC and CP development.
(A) Schematic representation of the npr3 gene structure showing the target site of Npr3 translation blocking morpholino (Npr3MO; red). (B) Western blot analysis of protein lysates from control …
Figure 2—figure supplement 1
Manipulation of the blastocoel fluid does not affect snai2 and six1 expression.
(A) The expression of snai2 and six1 is unaffected in embryos in which the blastocoel fluid was replaced by 100–120 nl of water at NF stage 9. Dorsal views, anterior to top. (B) Frequency of the …
Figure 2—figure supplement 2
Npr3-depletion affects the rate of cell death or proliferation in the dorsal ectoderm.
(A) Whole-mount TUNEL (left panel) and pHH3 (right panel) staining of stage 15 embryos injected with Npr3MO on the left side (asterisk). Dorsal views, anterior to top. The black boxes outline the …
Figure 3
Npr3 regulates NC and CP fate through different mechanisms.
(A) Two expression constructs were generated, a wild-type human NPR3 (HNPR3WT) and a truncated version of HNPR3 lacking the C-terminal Gi activator sequence (HNPR3ΔC). (B) Validation of the two …
Figure 4 with 1 supplement
Npr3 requires adenylyl cyclase inhibition to promote CP fate.
(A) Inhibition of adenylyl cyclase by treatment with SQ22536 restores six1 expression in Npr3-depleted (Npr3MO) embryos. Two representative embryos are shown to illustrate the range of phenotypes. …
Figure 4—figure supplement 1
Forskolin treatment interferes with six1 and eya1 induction by Zic1GR.
qRT-PCR analysis of six1 and eya1 expression in Zic1GR-injected animal caps untreated or treated with 50 μM Forskolin, from two biological replicates (A and B). Expression levels are normalized to od…
Figure 5 with 1 supplement
Signaling through Npr1 regulates NC and CP formation.
(A) Western blot analysis of protein lysates from control embryos and embryos injected with npr1 or npr2 mRNA (1 ng) alone or in combination with the corresponding translation blocking MO, Npr1MO …
Figure 5—figure supplement 1
Spatial expression of npr1, npr2, nppa and nppc by whole-mount ISH.
Spatial expression of npr1, npr2, nppa, and nppc by whole-mount ISH at neurula (NF stage 15/16) and tailbud (NF stage 28/32) stages. The embryonic stages are indicated in the lower right corner of …
Figure 6
Natriuretic peptides activity during NC and CP formation.
(A) ISH for snai2, sox10, six1 and sox2 expression in NppaMO-, NppbMO-, and NppcMO-injected embryos. The asterisks (*) mark the injected side, also visualize by the lineage tracer Red-gal. Dorsal …
Additional files
-
Transparent reporting form
- https://cdn.elifesciences.org/articles/84036/elife-84036-transrepform1-v2.pdf
-
Source data 1
- https://cdn.elifesciences.org/articles/84036/elife-84036-data1-v2.xlsx
