Deficiency of lrp4 in zebrafish and human LRP4 mutation induce aberrant activation of Jagged–Notch signaling in fin and limb development

Tian, Jing; Shao, Jinhui; Liu, Cong; Hou, Hsin-Yu; Shboul, Mohammad; Li, Guoqing; El-Khateeb, Mohammad; Samarah, Omar Q.; Kou, Yao; Chen, Yu-Hsuan; Chen, Mei-Jen; Lyu, Zhaojie; Chen, Wei-Leng; Sun, Yonghua; Liu, Yiwen

doi:10.1007/s00018-018-2928-3

Cited by 26 publications

(17 citation statements)

References 51 publications

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“…Total RNA was extracted from fresh zebrafish embryo tissues using TRIzol reagent (Ambion, Austin, TX, USA) as described in [29] with modifications. cDNA synthesis was performed using SuperScriptTM reverse transcriptase system (Invitrogen, Carlsbad, CA, USA).…”

Section: Real-time Quantitative Pcr (Qrt-pcr) Analysismentioning

confidence: 99%

“…Digoxigenin (DIG)-labeled riboprobes of cmlc1 were synthesized using RNA Labeling Kit (Roche, Basel, Switzerland). Whole-mount in situ hybridization (WISH) assays were performed as described previously [29,30]. Briefly, zebrafish embryos at different stages were collected and fixed in 4% paraformaldehyde at 4 • C overnight.…”

Section: Whole-mount In Situ Hybridization and Red Blood Cell Staininmentioning

confidence: 99%

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Generation and Application of the Zebrafish heg1 Mutant as a Cardiovascular Disease Model

Wang

et al. 2020

Biomolecules

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Cardiovascular disease (CVD) is the leading cause of global mortality, which has caused a huge burden on the quality of human life. Therefore, experimental animal models of CVD have become essential tools for analyzing the pathogenesis, developing drug screening, and testing potential therapeutic strategies. In recent decades, zebrafish has entered the field of CVD as an important model organism. HEG1, a heart development protein with EGF like domains 1, plays important roles in the development of vertebrate cardiovascular system. Loss of HEG1 will affect the stabilization of vascular endothelial cell connection and eventually lead to dilated cardiomyopathy (DCM). Here, we generated a heg1-specific knockout zebrafish line using CRISPR/Cas9 technology. Zebrafish heg1 mutant demonstrated severe cardiovascular malformations, including atrial ventricular enlargement, heart rate slowing, venous thrombosis and slow blood flow, which were similar to human heart failure and thrombosis phenotype. In addition, the expression of zebrafish cardiac and vascular markers was abnormal in heg1 mutants. In order to apply zebrafish heg1 mutant in cardiovascular drug screening, four Traditional Chinese Medicine (TCM) herbs and three Chinese herbal monomers were used to treat heg1 mutant. The pericardial area, the distance between sinus venosus and bulbus arteriosus (SV-BA), heart rate, red blood cells (RBCs) accumulation in posterior cardinal vein (PCV), and blood circulation in the tail vein were measured to evaluate the therapeutic effects of those drugs on DCM and thrombosis. Here, a new zebrafish model of DCM and thrombosis was established, which was verified to be suitable for drug screening of cardiovascular diseases. It provided an alternative method for traditional in vitro screening, and produced potential clinical related drugs in a rapid and cost-effective way.

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Section: Real-time Quantitative Pcr (Qrt-pcr) Analysismentioning

confidence: 99%

Section: Whole-mount In Situ Hybridization and Red Blood Cell Staininmentioning

confidence: 99%

Generation and Application of the Zebrafish heg1 Mutant as a Cardiovascular Disease Model

Wang

et al. 2020

Biomolecules

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“…So far, the understanding was that LRP4 is one of the member of canonical WNT pathways and its mutations may lead to dysregulation of WNT and BMP pathways, causing CLSS . However, a recent study has shown that a loss of function mutation (p.Arg373Trp) in EGF‐like 2 domain of LRP4 caused the induced activation of Jagged‐Notch signaling in the zebra fish fin and human limb development . The homozygous mutation, p.Tyr384Cys, as identified in the present study, lies in the EGF‐like 2 calcium binding domain of LRP4 protein and is found to be highly conserved among the species (Figure ).…”

Section: Discussionmentioning

confidence: 50%

Novel missense alteration in LRP4 gene underlies Cenani–Lenz syndactyly syndrome in a consanguineous family

Alrayes

Aziz²,

Ullah³

et al. 2020

The Journal of Gene Medicine

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Background: Syndactyly is a clinical feature of split-hand foot malformation (SHFM), ectodermal-dysplasia-syndactyly (EDSS1) and Cenani-Lenz syndactyly syndromes (CLSS). In EDSS1, only cutaneous syndactyly is observed, with sparse hair, abnormal nails and dentition. In SHFM, bony syndactyly may vary from hypoplasia of one phalanx to aplasia of central digits, extending to complete fusion of all fingers and toes in CLSS. Several genes have been assigned to these syndromes. Performing a single step molecular diagnostics becomes a challenge when a phenotype has overlaps with several syndromes or when some of the clinical features are not fully expressed in patients. Methods:Whole exome sequencing (WES) analysis on one sample derived from a consanguineous family was performed. A causative variant in WES data was prioritized via standard bioinformatics tools. The selected variant was Sanger sequenced in all the available family members for autosomal recessive segregation. Results:A novel missense variant (c.1151A>G; p.Tyr384Cys) was identified in the LRP4 gene. Sanger validation confirmed that all affected individuals were homozygous and the obligate carriers were heterozygous for this variant. The variant is neither reported in 1000 human genomes, nor in 60 706 exomes databases, and is predicted as "pathogenic" by SIFT, Polyphen-2 and MutationTaster software. Conclusions:The present study broadens the pathogenic spectrum of the LRP4 gene in syndactyly syndromes. WES is a powerful tool for genetic analysis in research and can be readily used as a first-line diagnostic test in syndactyly and related phenotypes. K E Y W O R D SCenani-Lenz syndactyly syndrome, LRP4, missense variant, WES analysis

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“…Therefore, AB staining has been largely used in early life stages, i.e., 2–20 dpf, to study the morphology of the chondrocranium in different skeletal zebrafish models ( 62 , 68 ) ( Figure 5 ). Developing malformations are mainly defined as the irregular shape of skeletal elements, but can also be defined by the absence of skeletal elements or the incorrect morphogenesis of a single skeletal element ( 66 , 84 ). Relative to the entire skeleton, not much cartilage is present in later life stages (late juveniles, adults) of zebrafish, yet AB staining can be used to assess for example cartilaginous joints ( 92 ).…”

Section: Bone Histology: From Whole Mount To Sectionsmentioning

confidence: 99%

Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

et al. 2020

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Animal models are essential tools for addressing fundamental scientific questions about skeletal diseases and for the development of new therapeutic approaches. Traditionally, mice have been the most common model organism in biomedical research, but their use is hampered by several limitations including complex generation, demanding investigation of early developmental stages, regulatory restrictions on breeding, and high maintenance cost. The zebrafish has been used as an efficient alternative vertebrate model for the study of human skeletal diseases, thanks to its easy genetic manipulation, high fecundity, external fertilization, transparency of rapidly developing embryos, and low maintenance cost. Furthermore, zebrafish share similar skeletal cells and ossification types with mammals. In the last decades, the use of both forward and new reverse genetics techniques has resulted in the generation of many mutant lines carrying skeletal phenotypes associated with human diseases. In addition, transgenic lines expressing fluorescent proteins under bone cell-or pathway-specific promoters enable in vivo imaging of differentiation and signaling at the cellular level. Despite the small size of the zebrafish, many traditional techniques for skeletal phenotyping, such as x-ray and microCT imaging and histological approaches, can be applied using the appropriate equipment and custom protocols. The ability of adult zebrafish to remodel skeletal tissues can be exploited as a unique tool to investigate bone formation and repair. Finally, the permeability of embryos to chemicals dissolved in water, together with the availability of large numbers of small-sized animals makes zebrafish a perfect model for high-throughput bone anabolic drug screening. This review aims to discuss the techniques that make zebrafish a powerful model to investigate the molecular and physiological basis of skeletal disorders.

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Deficiency of lrp4 in zebrafish and human LRP4 mutation induce aberrant activation of Jagged–Notch signaling in fin and limb development

Cited by 26 publications

References 51 publications

Generation and Application of the Zebrafish heg1 Mutant as a Cardiovascular Disease Model

Generation and Application of the Zebrafish heg1 Mutant as a Cardiovascular Disease Model

Novel missense alteration in LRP4 gene underlies Cenani–Lenz syndactyly syndrome in a consanguineous family

Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

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