ZFIN ID: ZDB-FIG-110531-28
Liu et al., 2011 - Targeting zebrafish and murine pituitary corticotroph tumors with a cyclin-dependent kinase (CDK) inhibitor. Proc. Natl. Acad. Sci. USA   108(20):8414-8419 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Term:
Stage: Long-pec
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Long-pec to Adult

Fig. 3 In vivo drug testing in Tg:Pomc-Pttg zebrafish. (A) Pttg overexpression directed by zebrafish Pomc promoter induced cyclin E up-regulation in Tg:Pomc-Pttg transgenic pituitary at 3 mo. mRNA levels were assayed by quantitative real-time PCR (mean ± SE of relative expression; n = 30 pituitaries for each group). (B) Western blot of mouse corticotroph tumor AtT20 cells transfected with a control or PTTG siRNA. (C) In vivo treatment of Tg:Pomc-Pttg;Pomc-eGFP embryos with small-molecule CDK inhibitors (50 μM) or 0.2% DMSO as control from 18 to 40 hpf. One hundred to one hundred fifty embryos were treated with each compound. Representative images of live embryos are shown with gross morphology (Right) and pituitary Pomc-GFP–positive cells at higher magnification (Left) at 40 hpf. Embryos exposed to flavopiridol developed early developmental defect before pituitary POMC cell ontogeny occurs. (D) Relative expression of pituitary Pomc-eGFP fluorescence analyzed using Volocity 5.2 software (Improvision; mean ± SE of relative expression, n = 7). (E) R-roscovitine specifically suppresses expansion of pituitary POMC cells overexpressing zPttg from 18 to 48 hpf. Double transgenic Tg:Pomc-Pttg;Prl-RFP embryos were generated by breeding Tg:Pomc-Pttg fish with a previously generated PRL-RFP transgenic line, in which RFP was targeted to pituitary lactotrophs by a zebrafish Prolactin promoter (34). Representative fluorescent microscopy of pituitary POMC-eGFP (a and b) and PRL-RFP (c and d) expression in live Tg:Pomc-Pttg;Pomc-eGFP and Tg:Pomc-Pttg;Prl-RFP embryos treated with 0.2% DMSO (a and c) or 50 μM R-roscovitine (b and d). (F) Relative expression of pituitary POMC-eGFP or PRL-RFP fluorescence were analyzed (mean ± SE of relative expression; n = 10). Results represent one of three similar experiments; *P < 0.02 and **P < 0.000005. (Scale bar, 50 μm.)

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP csz1Tg; zf44Tg control Long-pec rostral pars anterior IFL
csz1Tg; zf44Tg chemical treatment: seliciclib Long-pec rostral pars anterior IFL
RFP csz1Tg; zf113Tg control Long-pec rostral pars anterior IFL
csz1Tg; zf113Tg chemical treatment: seliciclib Long-pec rostral pars anterior IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
csz1Tg standard conditions Adult regulation of cyclin-dependent protein serine/threonine kinase activity decreased process quality, abnormal
csz1Tg; zf113Tg control Long-pec prolactin secreting cell amount, normal
csz1Tg; zf113Tg chemical treatment: seliciclib Long-pec prolactin secreting cell amount, normal
csz1Tg; zf44Tg control Long-pec adrenocorticotropic hormone secreting cell increased amount, abnormal
csz1Tg; zf44Tg chemical treatment: seliciclib Long-pec adrenocorticotropic hormone secreting cell amount, normal
Acknowledgments:
ZFIN wishes to thank the journal Proc. Natl. Acad. Sci. USA for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA