Visualization of skeletons and intervertebral disks in live fish larvae by fluorescent calcein staining and disk specific GFP expression

Haga, Yutaka; Du, Shao Jun; Masui, S.; Fujinami, Yuichiro; Aritaki, Masato; Satoh, Shuichi

doi:10.1111/j.1439-0426.2010.01418.x

Cited by 5 publications

(4 citation statements)

References 25 publications

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“…We used in vivo calcein staining, marking osteoblasts, and bone in live zebrafish (Du et al, 2001; Haga et al, 2010), to determine whether TCDD exposure inhibits ossification of the jaw cartilages. In control zebrafish embryos at 144 hpf, we found clear calcein staining in ceratohyal, dentary, and quadrate bone in whole mount live fluorescent images (Fig.…”

Section: Resultsmentioning

confidence: 99%

Dioxin disrupts cranial cartilage and dermal bone development in zebrafish larvae

2015

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2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD or dioxin) disrupts craniofacial development in zebrafish larvae. However, the cellular changes responsible for the decreased jaw size remain poorly understood. We show that smaller jaw size is due to a decrease in both the size and number of chondrocytes in the developing craniofacial cartilages. TCDD was found to decrease ossification of osteoblasts in the perichondrium of craniofacial cartilages. We also discovered that TCDD caused clefting of the parasphenoid, an effect with similarity to TCDD-induced cleft palate in mice. Thus, dermal and perichondrial bone development of the craniofacial skeleton are clearly disrupted by TCDD exposure in the zebrafish larvae. This dysmorphic response of the zebrafish craniofacial skeleton after exposure to TCDD is consistent with findings demonstrating disruption of axial bone development in medaka and repression of sox9b in zebrafish.

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Section: Resultsmentioning

confidence: 99%

Dioxin disrupts cranial cartilage and dermal bone development in zebrafish larvae

2015

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“…Because the notochord was present at fused centra at 53 dpf and had been degenerated at 121 dpf, it is hypothesized that the notochord gradually degenerates during larval to adult growth after fusion of centra. In zebrafish, it was also reported that RA exposure causes degeneration of notochord (Haga et al, 2010). Considering that the notochord cells produce extracellular matrices that form notochord sheath essential for intervertebral joint formation (Kaneko et al, 2016), it is presumed that degeneration of the notochord accelerates the progress of vertebral deformity.…”

Section: Discussionmentioning

confidence: 99%

Excess Retinoic Acid Induces Fusion of Centra by Degenerating Intervertebral Ligament Cells in Japanese flounder, Paralichthys olivaceus

Chen

Washio

et al. 2016

J Exp Zool Pt B

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In marine aquaculture fish, excessive supplement of vitamin A (VA) to zooplanktons for larval culture and experimental exposure of larvae to retinoic acid (RA: active form of VA) have been known to cause vertebral deformity. However, the tissues in the developing vertebral column that are affected by RA and the progression of vertebral deformity remain undetermined. To examine these questions, we histologically traced the progress of vertebral deformity induced by RA in Japanese flounder (Paralichthys olivaceus). Larvae were exposed to RA for 3 days at mid-metamorphosis (G-stage), a critical stage for vertebral deformity. Intervertebral ligament, which is known to form intervertebral joints in cooperation with the notochord, was severely degenerated by RA, leading to fusion of centra. During further development to adult, growth of centra was severely suppressed in an anterior-posterior direction in RA-treated fish and the notochord tissue was lost from fused centra, resulting in complete loss of intervertebral joints and fusion of centra. We conclude that RA initially damages the intervertebral ligaments, and these defects lead to fusion, narrowing of centra, and loss of intervertebral joints in the vertebral column. The cumulative effect of these modifications is a truncated body form.

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“…Fin rays were then fixed overnight in 10% buffered formalin, stepped into a 0.3% KOH solution, and mixed with 1–2 drops of 30% hydrogen peroxide per 100 ml of solution to remove skin pigmentation. Fin rays were then stained in calcein solution ( N = 2 for each species) following published methods (Haga et al, ) or cleared and stained ( N = 2 for each species) with alizarin red following published methods (Dingerkus & Uhler, ; Potthoff, ). Each fin ray's medio‐lateral surface was photographed under fluorescent light to aid in the visualization of segmentation.…”

Section: Methodsmentioning

confidence: 99%

“…Fin rays were then stained in calcein solution (N 5 2 for each species) following published methods (Haga et al, 2010) or cleared and stained (N 5 2 for each species) with alizarin red following published methods (Dingerkus & Uhler, 1977;Potthoff, 1984). Each fin ray's medio-lateral surface was photographed under fluorescent light to aid in the visualization of segmentation.…”

Section: Fin Ray Morphologymentioning

confidence: 99%

A comparison of pectoral fin ray morphology and its impact on fin ray flexural stiffness in labriform swimmers

Aiello

Hardy

Cherian

et al. 2018

Journal of Morphology

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The organization of tissues in appendages often affects their mechanical properties and function. In the fish family Labridae, swimming behavior is associated with pectoral fin flexural stiffness and morphology, where fins range on a continuum from stiff to relatively flexible fins. Across this diversity, pectoral fin flexural stiffness decreases exponentially along the length of any given fin ray, and ray stiffness decreases along the chord of the fin from the leading to trailing edge. In this study, we examine the morphological properties of fin rays, including the effective modulus in bending (E), second moment of area (I), segmentation, and branching patterns, and their impact on fin ray stiffness. We quantify intrinsic pectoral fin ray stiffness in similarly sized fins of two closely related species that employ fins of divergent mechanics, the flapping Gomphosus varius and the rowing Halichoeres bivittatus. While segmentation patterns and E were similar between species, measurements of I and the number of fin ray branch nodes were greater in G. varius than in H. bivittatus. A multiple regression model found that of these variables, I was always significantly correlated with fin ray flexural stiffness and that variation in I always explained the majority of the variation in flexural stiffness. Thus, while most of the morphological variables quantified in this study correlate with fin ray flexural stiffness, second moment of area is the greatest factor contributing to variation in flexural stiffness. Further, interspecific variation in fin ray branching pattern could be used as a means of tuning the effective stiffness of the fin webbing to differences in swimming behavior and hydrodynamics. The comparison of these results to other systems begins to unveil fundamental morphological features of biological beams and yields insight into the role of mechanical properties in fin deformation for aquatic locomotion.

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Visualization of skeletons and intervertebral disks in live fish larvae by fluorescent calcein staining and disk specific GFP expression

Cited by 5 publications

References 25 publications

Dioxin disrupts cranial cartilage and dermal bone development in zebrafish larvae

Dioxin disrupts cranial cartilage and dermal bone development in zebrafish larvae

Excess Retinoic Acid Induces Fusion of Centra by Degenerating Intervertebral Ligament Cells in Japanese flounder, Paralichthys olivaceus

A comparison of pectoral fin ray morphology and its impact on fin ray flexural stiffness in labriform swimmers

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