A two-color acid-free cartilage and bone stain for zebrafish larvae

Walker, Macie B.; Kimmel, Charles B.

doi:10.1080/10520290701333558

Cited by 649 publications

(513 citation statements)

References 12 publications

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“…After treatment, larvae were washed several times in fish water before returning to circulating culture flasks to be grown out to 12 dpf. Cartilage and bone staining was performed as previously described (47). Larvae were photographed with a Zeiss Axiocam digital imaging system mounted to an M2 Bio stereomicroscope (Zeiss).…”

Section: Methodsmentioning

confidence: 99%

Craniofacial divergence and ongoing adaptation via the hedgehog pathway

Roberts

Albertson

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

116

148

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Adaptive variation in craniofacial structure contributes to resource specialization and speciation, but the genetic loci that underlie craniofacial adaptation remain unknown. Here we show that alleles of the hedgehog pathway receptor Patched1 (Ptch1) gene are responsible for adaptive variation in the shape of the lower jaw both within and among genera of Lake Malawi cichlid fish. The evolutionarily derived allele of Ptch1 reduces the length of the retroarticular (RA) process of the lower jaw, a change predicted to increase speed of jaw rotation for improved suction-feeding. The alternate allele is associated with a longer RA and a more robustly mineralized jaw, typical of species that use a biting mode of feeding. Genera with the most divergent feeding morphologies are nearly fixed for different Ptch1 alleles, whereas species with intermediate morphologies still segregate variation at Ptch1. Thus, the same alleles that help to define macroevolutionary divergence among genera also contribute to microevolutionary fine-tuning of adaptive traits within some species. Variability of craniofacial morphology mediated by Ptch1 polymorphism has likely contributed to niche partitioning and ecological speciation of these fishes.T he astounding diversity of vertebrate craniofacial morphology reflects adaptation to a wide variety of resources and environments. Evolution of craniofacial structure has been key to niche specialization and speciation in vertebrates (1), most famously exemplified by the varied beak morphology of Darwin's finches (2). The radiation of finches demonstrates both speciesdefining craniofacial divergence (2) and rapid adaptation of craniofacial morphology within species over the course of a few years in response to changes in resource availability (3). Although shifts in the expression of key craniofacial genes have been correlated with differences in trophic morphology among species of birds and cichlid fishes (4-8), the specific genetic loci producing these differences remain uncharacterized. As a result, it is not clear whether interspecific divergence and intraspecific adaptation share the same molecular genetic basis.

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Section: Methodsmentioning

confidence: 99%

Craniofacial divergence and ongoing adaptation via the hedgehog pathway

Roberts

Albertson

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

116

148

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“…Skeletal staining was performed as described previously (17,32). In vivo skeletal staining was performed with 0.001% calcein or 0.05% alizarin red in E3 medium for 5-10 min and subsequent extensive washes with E3 medium.…”

Section: Methodsmentioning

confidence: 99%

Entpd5 is essential for skeletal mineralization and regulates phosphate homeostasis in zebrafish

Huitema

Apschner

Logister

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

100

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Bone mineralization is an essential step during the embryonic development of vertebrates, and bone serves vital functions in human physiology. To systematically identify unique gene functions essential for osteogenesis, we performed a forward genetic screen in zebrafish and isolated a mutant, no bone (nob), that does not form any mineralized bone. Positional cloning of nob identified the causative gene to encode ectonucleoside triphosphate/ diphosphohydrolase 5 (entpd5); analysis of its expression pattern demonstrates that entpd5 is specifically expressed in osteoblasts. An additional mutant, dragonfish (dgf), exhibits ectopic mineralization in the craniofacial and axial skeleton and encodes a loss-offunction allele of ectonucleotide pyrophosphatase phosphodiesterase 1 (enpp1). Intriguingly, generation of double-mutant nob/ dgf embryos restored skeletal mineralization in nob mutants, indicating that mechanistically, Entpd5 and Enpp1 act as reciprocal regulators of phosphate/pyrophosphate homeostasis in vivo. Consistent with this, entpd5 mutant embryos can be rescued by high levels of inorganic phosphate, and phosphate-regulating factors, such as fgf23 and npt2a, are significantly affected in entpd5 mutant embryos. Our study demonstrates that Entpd5 represents a previously unappreciated essential player in phosphate homeostasis and skeletal mineralization.

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“…We prepared post-hatching skeletons with standard clearing and staining protocols [49][50][51]. Scaled digital photographs of the limb elements were obtained using a Nikon DN100 camera mounted on a Nikon SMZ1000 dissecting microscope.…”

Section: Methodsmentioning

confidence: 99%

Repeated modification of early limb morphogenesis programmes underlies the convergence of relative limb length inAnolislizards

et al. 2011

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The independent evolution of similar morphologies has long been a subject of considerable interest to biologists. Does phenotypic convergence reflect the primacy of natural selection, or does development set the course of evolution by channelling variation in certain directions? Here, we examine the ontogenetic origins of relative limb length variation among Anolis lizard habitat specialists to address whether convergent phenotypes have arisen through convergent developmental trajectories. Despite the numerous developmental processes that could potentially contribute to variation in adult limb length, our analyses reveal that, in Anolis lizards, such variation is repeatedly the result of changes occurring very early in development, prior to formation of the cartilaginous long bone anlagen.

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A two-color acid-free cartilage and bone stain for zebrafish larvae

Cited by 649 publications

References 12 publications

Craniofacial divergence and ongoing adaptation via the hedgehog pathway

Craniofacial divergence and ongoing adaptation via the hedgehog pathway

Entpd5 is essential for skeletal mineralization and regulates phosphate homeostasis in zebrafish

Repeated modification of early limb morphogenesis programmes underlies the convergence of relative limb length inAnolislizards

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