Yildiz et al., 2019 - Zebrafish prdm12b acts independently of nkx6.1 repression to promote eng1b expression in the neural tube p1 domain. Neural Development   14:5 Full text @ Neural Dev.

Fig. 2

prdm12b germ line mutants lack eng1b expression in the p1 domain. a, b. prdm12b is not maternally deposited. In situ hybridization detects prdm12b expression at 24hpf (b), but not at 2.5hpf (a), in wildtype embryos. c. Bar chart depicting the frequency of each genotype at various time points in broods from crosses of prdm12b heterozygous animals. Error bars indicate ±S. E. (n = 3). dpf = days post fertilization, mo = months. d, e. Morphology of 15dpf prdm12b+/+ (d) and prdm12bum319/um319 (e) fish. f-s. eng1b expression in 24hpf embryos from crosses of prdm12b+/um318 heterozygotes (f-k), or prdm12b+/um319 heterozygotes (l-s). Numbers in each panel indicate the fraction of animals with the specified phenotype. t, u. evx1expression in 24hpf embryos from a cross of prdm12b+/um319 heterozygotes. v, w. vsx2expression in 24hpf embryos from a cross of prdm12b+/um319 heterozygotes. Embryos are shown in dorsal (f-h, l-n, t-y) or lateral (i-k, o-q) view with anterior to the left, or in cross section (r, s) with dorsal to the top. Brackets indicate r4, arrows mark V1 interneurons and arrowheads mark somites. MHB = midbrain–hindbrain boundary, HB = hindbrain and SC = spinal cord

Fig. 3

prdm12b mutant fish display an abnormal touch evoked response. a-d. Representative kinematic traces for 10 wildtype (a) and 11 prdm12b mutant (b) fish stimulated with a head touch, as well as for 11 prdm12b mutants first assayed with a head touch (c) and subsequently with a tail touch (d). Zero degrees on the y-axis indicate a straight body while positive and negative angles represent body bends in opposite directions. All fish were at 4dpf. e. Anti-3A10 labeling of Mauthner neurons in a cross of prdm12b+/um318 heterozygotes (n = 117). f. Anti-F310 labeling of somites in a cross of prdm12b+/um398 heterozygotes (n = 16)

Fig. 6

Analysis of bhlhe22 mutant zebrafish. a. Chart depicting the frequency of each genotype at various timepoints in broods from crosses of bhlhe22+/um320 heterozygous fish. mo = month, y/o = year old. b. Sequencing traces of transcripts from wild type versus bhlhe22um320/um320animals showing the expected 5 bp deletion. c-e. Expression of eng1b (c), evx1 (d) and vsx2(e) in 24hpf wildtype and bhlhe22um320/um320 mutant embryos

Fig. 7

prdm12b does not maintain the p1 domain by repressing nkx6.1. a. Anti-Nkx6.1 immunostaining of nkx6.1um321/um321 mutant (left) and wildtype (right) embryos at 30hpf. b. Chart indicating the frequency of each genotype at various time points in broods from crosses of nkx6.1+/um321 heterozygous mutants. c. Hb9 immunostaining in wildtype (left) versus a cross of nkx6.1+/um321 heterozygous embryos (right) at 33hpf. d. Islet-1/2 immunostaining of 50hpf embryos from a cross of nkx6.1+/um321 heterozygotes. e. Expression of eng1b in 24hpf embryos from a cross of nkx6.1+/um321 heterozygotes. f. Expression of eng1b in 24hpf uninjected wildtype embryos (left panels), 24hpf prdm12b MO-injected wildtype embryos (middle panels) and 24hpf prdm12b MO-injected embryos from a cross of nkx6.1+/um321heterozygotes (right panels). g. Expression of prdm12b in a representative wildtype embryo (left panel) and a representative embryo from a cross of nkx6.1+/um321 heterozygotes (middle panel) at 24hpf. Right panel shows quantification of the size of the prdm12b expression domain in 11 wildtype embryos and 20 embryos from a cross of nkx6.1+/um321 heterozygotes. Numbers in panels indicate the fraction of embryos displaying the phenotype shown

Fig. 8

prdm12b controls the size of the p1 domain. Expression of pax3/nkx6.1 (a, b), dbx1/nkx6.1 (d, e), pax3/vsx2 (g, h), dbx1/vsx2 (j, k) and evx1/nkx6.1 (m, n) in 24hpf wildtype (a, d, g, j, m) or prdm12b MO-injected (b, e, h, k, n) embryos. Panels show cross sections through the spinal cord with dorsal to the top. c, f, i, l, o show quantification of the size (along the dorsoventral axis) of the p0/p1 domain (c, i) or the p1 domain (f, l, o) relative to the neural tube. At least 10 representative sections were used for each gene pair

PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Prim-5

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EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Terms:
Stage: Prim-5
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Neural Dev.