[Show abstract][Hide abstract] ABSTRACT: The categorical data set is an important data class in experimental biology and contains data separable into several mutually exclusive categories. Unlike measurement of a continuous variable, categorical data cannot be analyzed with methods such as the Student's t-test. Thus, these data require a different method of analysis to aid in interpretation. In this article, we will review issues related to categorical data, such as how to plot them in a graph, how to integrate results from different experiments, how to calculate the error bar/region, and how to perform significance tests. In addition, we illustrate analysis of categorical data using experimental results from developmental biology and virology studies.
[Show abstract][Hide abstract] ABSTRACT: Cilia defects have been implicated in a variety of human diseases and genetic disorders, but how cilia motility contributes to these phenotypes is still unknown. To further our understanding of how cilia function in development, we have cloned and characterized two alleles of seahorse, a zebrafish mutation that results in pronephric cysts. seahorse encodes Lrrc6l, a leucine-rich repeat-containing protein that is highly conserved in organisms that have motile cilia. seahorse is expressed in zebrafish tissues known to contain motile cilia. Although mutants do not affect cilia structure and retain the ability to interact with Disheveled, both alleles of seahorse strongly affect cilia motility in the zebrafish pronephros and neural tube. Intriguingly, although seahorse mutations variably affect fluid flow in Kupffer's vesicle, they can have very weak effects on left-right patterning. Combined with recently published results, our alleles suggest that the function of seahorse in cilia motility is separable from its function in other cilia-related phenotypes.
Development 06/2009; 136(10):1621-31. · 6.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Work in mouse has implicated cilia motility and leftward nodal flow as the mechanism for breaking left-right symmetry. In zebrafish, it is assumed that Kupffer's vesicle is analogous to the mouse node. However, its architecture is different and the fluid dynamics inside Kupffer's vesicle is not completely understood. We show that cells lining both the dorsal roof and the ventral floor of Kupffer's vesicle possess posteriorly pointed cilia that rotate clockwise when viewed apically. Analysis of bead movements within Kupffer's vesicle shows a net circular flow but the local flow differs in direction depending on the location within the vesicle. Histological analysis suggests that the orientation of the cells at anterior-dorsal region likely direct net flow in the vesicle. Our data suggest that the plane of the circular net flow is tilted with respect to the D-V axis, which may be converted to a local leftward flow in the anterior-dorsal region of the vesicle.
[Show abstract][Hide abstract] ABSTRACT: In many vertebrates, extra-embryonic tissues are important signaling centers that induce and pattern the germ layers. In teleosts, the mechanism by which the extra-embryonic yolk syncytial layer (YSL) patterns the embryo is not understood. Although the Nodal-related protein Squint is expressed in the YSL, its role in this tissue is not known. We generated a series of stable transgenic lines with GFP under the control of squint genomic sequences. In all species, nodal-related genes induce their own expression through a positive feedback loop. We show that two tissue specific enhancers in the zebrafish squint gene mediate the response to Nodal signals. Expression in the blastomeres depends upon a conserved Nodal response element (NRE) in the squint first intron, while expression in the extra-embryonic enveloping layer (EVL) is mediated by an element upstream of the transcription start site. Targeted depletion experiments demonstrate that the zebrafish Nodal-related proteins Squint and Cyclops are required in the YSL for endoderm and head mesoderm formation. Thus, Nodal signals mediate interactions between embryonic and extra-embryonic tissues in zebrafish that maintain nodal-related gene expression in the margin. Our results demonstrate a high degree of functional conservation between the extra-embryonic tissues of mouse and zebrafish.