Screening mosaic F1 females for mutations affecting zebrafish heart induction and patterning.
The genetic pathways underlying the induction and anterior-posterior patterning of the heart are poorly understood. The recent emergence of the zebrafish model system now allows a classical genetic approach to such challenging problems in vertebrate development. Two large-scale screens for mutations affecting zebrafish embryonic development have recently been completed; among the hundreds of mutations identified were several that affect specific aspects of cardiac morphogenesis, differentiation, and function. However, very few mutations affecting induction and/or anterior-posterior patterning of the heart were identified. We hypothesize that a directed approach utilizing molecular markers to examine these particular steps of heart development will uncover additional such mutations. To test this hypothesis, we are conducting two parallel screens for mutations that affect either the induction or the anterior-posterior patterning of the zebrafish heart. As an indicator of cardiac induction, we examine expression of nkx2.5, the earliest known marker of precardiac mesoderm; to assess anterior-posterior patterning, we distinguish ventricle from atrium with antibodies that recognize different myosin heavy chain isoforms. In order to expedite the examination of a large number of mutations, we are screening the haploid progeny of mosaic F1 females. In these ongoing screens, we have identified four mutations that affect nkx2.5 expression as well as 21 that disrupt either ventricular or atrial development and thus far have recovered several of these mutations, demonstrating the value of our approach. Future analysis of these and other cardiac mutations will provide further insight into the processes of induction and anterior-posterior patterning of the heart.
Alexander
J
J
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0554, USA.
Stainier
D Y
DY
Yelon
D
D
D000818:Animals; D019521:Body Patterning; D002454:Cell Differentiation / Q000502:physiology; D002999:Clone Cells; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D005819:Genetic Markers; D005820:Genetic Testing; D006238:Haploidy; D006321:Heart / Q000196:embryology*; D008297:Male; D009024:Morphogenesis; D009030:Mosaicism; D009154:Mutation; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*
D005819:Genetic Markers