ZFIN ID: ZDB-FIG-210128-26
Petratou et al., 2021 - The MITF paralog tfec is required in neural crest development for fate specification of the iridophore lineage from a multipotent pigment cell progenitor. PLoS One   16:e0244794 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig 5 Specification of melanocytes, iridophores and xanthophores is delayed in <italic>tfec</italic> mutants.

(A-F) pnp4a expression in Ib(sp) requires Tfec. At 24 hpf, the number of dorsally located, and of partially fate restricted, migrating chromatoblasts (arrows) expressing pnp4a is reduced in tfec mutants (A,B). Expression overlying the eye is also affected (A,B, insets). This reduction is still prominent at 30 hpf (C,D), when staining is also visibly affected in the lateral patches. pnp4a expression is undetectable in differentiated iridophore positions (E, asterisks) in tfec mutants at 48 hpf (F). pnp4a+ iridophore lineage cells overlying the RPE are noticeably reduced in tfec mutants compared to WT siblings (A-F, insets). (G-L) Generation of tfec-positive Ib(sp) from late Cbls requires Tfec. At 24 hpf, tfec mutants present with anterior expansion of the tfec+ premigratory NC domain (G,H, arrowheads) and lack of medially migrating, Ib(sp) progenitors (arrows). At 36 hpf, the number of dorsal and migrating tfec+ Ib(df) (arrows), as well as those located in the lateral patches is reduced in mutants, compared to WT or heterozygous siblings (I,J). At 48 hpf, a reduced number of cells (asterisks) express tfec along the dorsal stripe of mutant embryos, compared to WT or heterozygous siblings (K,L). tfec mutant embryos were distinguishable after whole-mount in situ hybridisation by the lack of RPE melanisation (I-L, insets). tfec+ iridophore lineage cells overlying the RPE are noticeably reduced in tfec mutants compared to WT siblings (G-L, insets). (M-R) Mb(sp) generation from the late Cbl is delayed in tfec mutants. At 24 hpf, mitfa marker expression is restricted to an increased number of premigratory Cbls (arrowheads) and is undetectable in migrating Mb(sp) (arrows) in tfec mutants (M,N). At 30 hpf, the numbers and distributions of mitfa-positive late Cbls (arrowheads) are indistinguishable between tfec mutants and WT or heterozygous siblings (O,P), but the delay in Mb(sp) migration (arrows) remains distinguishable towards the tail. tfec mutant embryos lack RPE melanisation (O,P, insets). At 48 hpf, mitfa expression in mature melanocytes (asterisks) is indistinguishable between tfec mutants and WT or heterozygous siblings (Q,R). (S, T) tfec mutants lack ltk expression in premigratory late Cbls (arrowhead), in migrating Ib(sp) (arrow) and in iridoblasts of the eye (insets). (U-X) Xanthoblast (Xbl(sp)) specification from Cbls is delayed, as indicated by examinations of two lineage markers, gch2 (U,V) and aox5 (W,X). At 24 hpf, laterally migrating Xbl(sp) (arrows) are restricted to more anterior regions and precursors located in the head are reduced in tfec mutants (V,X) compared to their WT siblings (U,W). All tfec mutant panels show the tfecba6 allele except (V,X) which show the tfecvc60 allele. LP, lateral patches; m, melanisation. (A-J,M-X): lateral views. (K-L, insets of Q-R): dorsal views. Head towards the left. Scale bars: 100 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
aox5 WT standard conditions Prim-5 xanthoblast ISH
tfecvc60/vc60 standard conditions Prim-5 xanthoblast ISH
gch2 WT standard conditions Prim-5 xanthoblast ISH
tfecvc60/vc60 standard conditions Prim-5 xanthoblast ISH
ltk WT standard conditions Prim-5 iridoblast ISH
tfecba6/ba6 standard conditions Prim-5 Not Detected iridoblast ISH
mitfa WT standard conditions Prim-5 melanoblast ISH
Prim-15 melanoblast ISH
Long-pec forebrain ISH
Long-pec melanocyte ISH
tfecba6/ba6 standard conditions Prim-5 Not Detected melanoblast ISH
Prim-15 melanoblast ISH
Long-pec melanocyte ISH
pnp4a WT standard conditions Prim-5 iridoblast ISH
Prim-15 iridoblast ISH
Long-pec iridophore ISH
tfecba6/ba6 standard conditions Prim-5 iridoblast ISH
Prim-15 iridoblast ISH
Long-pec Not Detected iridophore ISH
tfec WT standard conditions Prim-5 premigratory neural crest ISH
Prim-25 iridoblast ISH
Long-pec iridophore ISH
tfecba6/ba6 standard conditions Prim-5 premigratory neural crest ISH
Prim-25 iridoblast ISH
Long-pec dorsal larval melanophore stripe iridophore ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
tfecba6/ba6 standard conditions Prim-5 iridoblast ltk expression absent, abnormal
Prim-5 iridoblast pnp4a expression decreased amount, abnormal
Prim-5 melanoblast mitfa expression absent, abnormal
Prim-15 iridoblast pnp4a expression decreased amount, abnormal
Prim-15 melanoblast mitfa expression decreased amount, abnormal
Prim-25 iridoblast tfec expression decreased amount, abnormal
Long-pec dorsal larval melanophore stripe iridophore tfec expression decreased amount, abnormal
Long-pec iridophore pnp4a expression absent, abnormal
Long-pec melanocyte mitfa expression amount, ameliorated
tfecvc60/vc60 standard conditions Prim-5 xanthoblast aox5 expression decreased distribution, abnormal
Prim-5 xanthoblast gch2 expression decreased distribution, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One