[Forward genetic screening for zebrafish mutants defective in erythropoiesis].
OBJECTIVE
To screen and identify zebrafish mutants with erythropoiesis defects by N-ethyl-N-nitrosourea (ENU) mutagenesis and large-scale forward genetic screening using beta e 1 as the marker.


METHODS
The chemical mutagen ENU was used to treat healthy wild-type male fish (AB strain, F0). The surviving ENU-treated fish were mated with wild-type female fish to generate F1, and further F2 family was generated by F1 family intercross. The adult F2 fish were intercrossed within each F2 family and the resulting F3 embryos from each crossing were subjected to whole mount in situ hybridization (WISH) with the beta e 1 probe. Mutagenesis was performed by treating the male zebrafish with ENU to induce mutations in pre-meiotic germ cells to generate the founders, which were outcrossed to obtained the F1 fish. The F1 fish from different founders were mated to generate the F2 families. F3 embryos from the sibling cross in the F2 family were examined by whole mount in situ hybridization using beta e 1-globin probe. The putative mutants were then characterized with different hematopoiesis markers.


CONCLUSIONS
We identified 4 beta e 1-deficient mutants with erythropoiesis defects, including two with specific erythiod lineage defects and two with concurrent lymphopoiesis defects.
Huo
Zhong-jun
ZJ
Department of Cell Biology, Southern Medical University, Guangzhou 510515, China.
Wen
Zong-hua
ZH
Lin
Jing
J
Wang
Kun
K
Huang
Zhi-bin
ZB
Dai
Zhao-xia
ZX
Ma
Ning
N
Yan
Guang
G
Chen
Ying-hua
YH
Chen
Xiao-hui
XH
Liu
Wei
W
Ma
Pin-yun
PY
Luo
Wei-hao
WH
Zhao
Ying
Y
Fan
Shu
S
Zhao
Jia-jia
JJ
Huang
Hong-hui
HH
Wen
Zi-long
ZL
Zhang
Wen-qing
WQ
D000818:Animals; D004920:Erythropoiesis / Q000235:genetics*; D005038:Ethylnitrosourea; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008297:Male; D016254:Mutagenesis, Insertional; D009154:Mutation; D015027:Zebrafish / Q000235:genetics*
D005038:Ethylnitrosourea