ZFIN ID: ZDB-FIG-150318-33
Koch et al., 2014 - A Mutation in cnot8, Component of the Ccr4-Not Complex Regulating Transcript Stability, Affects Expression Levels of Developmental Regulators and Reveals a Role of Fgf3 in Development of Caudal Hypothalamic Dopaminergic Neurons. PLoS One   9:e113829 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Protruding-mouth
PHENOTYPE:
Fish:
Observed In:
Stage Range: Protruding-mouth to Day 4

Fig. 1 m1061 mutants show increased th expression and increase in number of hypothalamic DA neurons from 3 dpf onwards.

Analysis of th gene expression in cnot8m1061 mutants and WT siblings as indicated in header. (A–F) th transcript levels are not altered in m1061/m1061 mutants at 1 and 2 dpf. Embryos were genotyped by PCR. (G–N) m1061 mutants at 3 dpf display stronger WISH signal indicating increased th mRNA levels in DA groups 1 to 7 (arrowhead) and NA sympathetic cells (asterisk). (K, L) NA locus coeruleus (LC) and DA pretectal cells (Pre) do not display altered th expression levels in m1061 mutant embryos. (M, N) The number of DA retinal amacrine cells (rAC) is reduced in m1061 mutant embryos. Additionally m1061 mutants display a lens defect (arrowheads). Genotypes were inferred by th WISH analysis. (O, P) anti-TH immunohistochemistry of m1061 mutant embryo and WT sibling at 4 dpf document DA neurons in the ventral diencephalon. Data confirm a higher cell number of DC7 DA neurons in the caudal hypothalamus of m1061 mutants. Dorsal view z-projections of partial confocal stacks representing the ventral diencephalon are shown. (A, B, E, F, G, H, M, N) lateral views, anterior at left; (C, D, G, H, K.L, O, P) dorsal views, anterior at left. Scale bars 100 µm in A for A, B; in C for C–F; in G for G–N; in P for O, P. Abbreviations: DC - early diencephalic DA group (DC2, DC4), 1 - ventral thalamic DA group, 2,4,5,6 posterior tubercular and hypothalamic Orthopedia-dependent DA groups, 3 - medial hypothalamic DA group, 7 - caudal hypothalamic DA group, VT - ventral thalamus, PT/H - posterior tuberculum/hypothalamus, PO - preoptic area, AAC - arch associated catecholaminergic neurons/carotid body, sym - sympathetic NA neurons, Pr - pretectum, TC - telencephalon, CH - caudal hypothalamus, LC locus coeruleus, MO - medulla oblongata, rAC - retinal amacrine cells. (Q) Quantification of CA cell numbers in m1061 embryos at 4 dpf. Cell counts of DC1-7 and LC TH-expressing cells. Bars show the average number of CA neurons in five m1061 mutants and five WT sibling embryos. Error bars indicate standard deviation. Significance was evaluated using Mann-Whitney test (see text and Table S1).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
th WT standard conditions Prim-5 diencephalon dopaminergic neuron ISH
Long-pec caudal tuberculum ISH
Long-pec caudal tuberculum dopaminergic neuron ISH
Long-pec hypothalamus ISH
Long-pec hypothalamus dopaminergic neuron ISH
Long-pec locus coeruleus ISH
Long-pec preoptic area ISH
Long-pec ventral thalamus ISH
Long-pec ventral thalamus dopaminergic neuron ISH
Protruding-mouth amacrine cell ISH
Protruding-mouth caudal tuberculum ISH
Protruding-mouth caudal tuberculum dopaminergic neuron ISH
Protruding-mouth hypothalamus ISH
Protruding-mouth hypothalamus dopaminergic neuron ISH
Protruding-mouth locus coeruleus ISH
Protruding-mouth medulla oblongata ISH
Protruding-mouth pretectum ISH
Protruding-mouth sympathetic neuron ISH
Protruding-mouth telencephalon ISH
Protruding-mouth ventral thalamus ISH
Protruding-mouth ventral thalamus dopaminergic neuron ISH
cnot8m1061/m1061 standard conditions Prim-5 diencephalon dopaminergic neuron ISH
Long-pec caudal tuberculum ISH
Long-pec caudal tuberculum dopaminergic neuron ISH
Long-pec hypothalamus ISH
Long-pec hypothalamus dopaminergic neuron ISH
Long-pec locus coeruleus ISH
Long-pec preoptic area ISH
Long-pec ventral thalamus ISH
Long-pec ventral thalamus dopaminergic neuron ISH
Protruding-mouth amacrine cell ISH
Protruding-mouth caudal tuberculum ISH
Protruding-mouth caudal tuberculum dopaminergic neuron ISH
Protruding-mouth hypothalamus ISH
Protruding-mouth hypothalamus dopaminergic neuron ISH
Protruding-mouth locus coeruleus ISH
Protruding-mouth medulla oblongata ISH
Protruding-mouth pretectum ISH
Protruding-mouth sympathetic neuron ISH
Protruding-mouth telencephalon ISH
Protruding-mouth ventral thalamus ISH
Protruding-mouth ventral thalamus dopaminergic neuron ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
cnot8m1061/m1061 standard conditions Protruding-mouth lens shape, abnormal
Day 4 hypothalamus posterior side has extra parts of type dopaminergic neuron, 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