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Grafting of Beads into Developing Chicken Embryo Limbs to Identify Signal Transduction Pathways Affecting Gene Expression

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miR-196 regulates axial patterning and pectoral appendage initiation.

Xinjun He¹, Yi-Lin Yan, Johann K Eberhart

¹Institute of Neuroscience, University of Oregon, Eugene, OR 97403, USA. xhe@uoneuro.uoregon.edu

Developmental Biology

|July 27, 2011

Summary

MicroRNA miR-196, found in Hox clusters, influences zebrafish development. It affects body axis patterning and limb development, partly by regulating retinoic acid signaling.

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Area of Science:

Developmental Biology
Genetics
Molecular Biology

Background:

Vertebrate Hox clusters encode protein-coding genes and microRNAs (miRNAs).
The function of Hox cluster-encoded miRNAs, like miR-196, in development is largely unknown.
Hox genes are crucial for body axis formation.

Purpose of the Study:

To investigate the developmental roles of the Hox cluster microRNA, miR-196, in zebrafish.
To elucidate the molecular mechanisms underlying miR-196's function in axial and appendicular patterning.

Main Methods:

Overexpression and knockdown of miR-196 in zebrafish embryos.
Phenotypic analysis of zebrafish development.
Gene expression analysis of Hox genes and retinoic acid signaling pathway components.
Reporter assays to identify miR-196 targets.

Main Results:

miR-196 overexpression caused pectoral fin defects, pharyngeal arch deletion, vertebral/rib anomalies, and somite reduction.
miR-196 knockdown led to extra pharyngeal arches, ribs, and somites, confirming endogenous roles.
miR-196 modulated Hox gene expression and retinoic acid signaling via the rarab gene.
The rarab 3'UTR was identified as a direct miR-196 target crucial for pectoral fin development.

Conclusions:

Hox cluster-derived miR-196 plays significant roles in both axial and appendicular patterning in zebrafish.
miR-196 functions similarly to protein-coding Hox genes in axial patterning.
miR-196 regulates appendicular patterning, at least partially, by modulating retinoic acid signaling through targeting rarab.