Lasp1 Expression Is Implicated in Embryonic Development of Zebrafish

Grossi, Ilaria; Schiavone, Marco; Cannone, Elena; Grejdan, Oana Andreea; Tobia, Chiara; Bonomini, Francesca; Rezzani, Rita; Salvi, Alessandro; Petro, Giuseppina De

doi:10.3390/genes14010035

Cited by 5 publications

(4 citation statements)

References 34 publications

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“…As shown in Figure 2, more than 50% of Neu3.2-MO embryos showed severe phenotypes at 24 hpf and this value further increased at 72 hpf, with the remaining morphants being affected by a mild one. To avoid the non-specific activation of p53 [10], a p53-MO was co-injected with neu3.2-MO, and the phenotypes observed were similar to those obtained after the co-injections of neu3.2-MO alone (not shown). These data indicate that phenotypes induced by neu3.2-MO were not due to the non-specific activation of p53 gene.…”

Section: Silencing Of Neu32 By Morpholino Leads To Morphology Alterat...mentioning

confidence: 55%

Downregulation of Zebrafish Cytosolic Sialidase Neu3.2 Affects Skeletal Muscle Development

Zizioli,

Codenotti,

Benaglia

et al. 2023

IJMS

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Sialidases remove terminal sialic acids residues from the non-reducing ends of glycoconjugates. They have been recognized as catabolic enzymes that work within different subcellular compartments and can ensure the proper turn-over of glycoconjugates. Four mammalian sialidases (NEU1-4) exist, with different subcellular localization, pH optimum and substrate specificity. In zebrafish, seven different sialidases, with high homology to mammalian counterparts, have been identified. Zebrafish Neu3.2 is similar to the human cytosolic sialidase NEU2, which is involved in skeletal muscle differentiation and exhibits a broad substrate specificity toward gangliosides and glycoproteins. In zebrafish neu3.2, mRNA is expressed during somite development, and its enzymatic activity has been detected in the skeletal muscle and heart of adult animals. In this paper, 1–4-cell-stage embryos injected with neu3.2 splice-blocking morpholino showed severe embryonic defects, mainly in somites, heart and anterior–posterior axis formation. Myog and myod1 expressions were altered in morphants, and impaired musculature formation was associated with a defective locomotor behavior. Finally, the co-injection of Neu2 mouse mRNA in morphants rescued the phenotype. These data are consistent with the involvement of cytosolic sialidase in pathologies related to muscle formation and support the validity of the model to investigate the pathogenesis of the diseases.

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Section: Silencing Of Neu32 By Morpholino Leads To Morphology Alterat...mentioning

confidence: 55%

Downregulation of Zebrafish Cytosolic Sialidase Neu3.2 Affects Skeletal Muscle Development

Zizioli,

Codenotti,

Benaglia

et al. 2023

IJMS

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“…The MS-ddPCR assays were optimized according to the principles of MS-PCR ( 34 , 35 ) to detect the methylation levels of SEPT9 and SHOX2 . MS-ddPCR experiments were performed using QX200™ ddPCR System (Bio-Rad Laboraties, Inc.) ( 36 , 37 ). The MS-ddPCR reaction mix consisted of the 2X ddPCR Supermix for Probes, and locus-specific primers and probes.…”

Section: Methodsmentioning

confidence: 99%

Section: Plasmamentioning

confidence: 99%

Evaluation of DNA methylation levels of SEPT9 and SHOX2 in plasma of patients with head and neck squamous cell carcinoma using droplet digital PCR

Grossi,

Assoni,

Lorini

et al. 2024

Oncol Rep

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Head and neck squamous cell carcinoma (HNSCC) is the seventh most commonly diagnosed cancer globally. HNSCC develops from the mucosa of the oral cavity, pharynx and larynx. Methylation levels of septin 9 ( SEPT9 ) and short stature homeobox 2 ( SHOX2 ) genes in circulating cell-free DNA (ccfDNA) are considered epigenetic biomarkers and have shown predictive value in preliminary reports in HNSCC. Liquid biopsy is a non-invasive procedure that collects tumor-derived molecules, including ccfDNA. In the present study, a droplet digital PCR (ddPCR)-based assay was developed to detect DNA methylation levels of circulating SEPT9 and SHOX2 in the plasma of patients with HNSCC. The assay was first set up using commercial methylated and unmethylated DNA. The dynamic changes in the methylation levels of SEPT9 and SHOX2 were then quantified in 20 patients with HNSCC during follow-up. The results highlighted: i) The ability of the ddPCR-based assay to detect very low copies of methylated molecules; ii) the significant decrease in SEPT9 and SHOX2 methylation levels in the plasma of patients with HNSCC at the first time points of follow-up with respect to T 0 ; iii) a different trend of longitudinally DNA methylation variations in small groups of stratified patients. The absolute and precise quantification of SEPT9 and SHOX2 methylation levels in HNSCC may be useful for studies with translational potential.

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“…Droplet Digital PCR Workflow QX200 Droplet Digital PCR System (Bio-Rad Laboratories) was employed to carry out this experiment and the synthesized cDNA was used as a template for the ddPCR experiments. ddPCR was performed according to the ddPCR Supermix for Probes (Bio-Rad Laboratories) protocol, as previously described [67]. Briefly, 1.33 µL of the cDNA obtained using TaqMan microRNA Reverse Transcription Kit, were prepared for amplification in a 20 µL reaction volume containing 2× ddPCR Supermix for Probes (Bio-Rad Laboratories), 20× TaqMan assay (Thermo Fisher Scientific) specific for miR-21-5p, miR-23b-3p, miR-24-3p, miR-27a-3p, miR-34a-3p, miR-126-3p, miR-133a-3p, miR-193a-3p, and water.…”

Section: Expression Of Predicted Mirnas With Rt-qpcrmentioning

confidence: 99%

Dysregulation of a Subset of Circulating and Vesicle-Associated miRNA in Pancreatic Cancer

Girolimetti,

Pelisenco,

Eusebi

et al. 2024

ncRNA

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Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive neoplasia, characterized by early metastasis, low diagnostic rates at early stages, resistance to drugs, and poor prognosis. There is an urgent need to better characterize this disease in order to identify efficient diagnostic/prognostic biomarkers. Since microRNAs (miRNAs) contribute to oncogenesis and metastasis formation in PDAC, they are considered potential candidates for fulfilling this task. In this work, the levels of two miRNA subsets (involved in chemoresistance or with oncogenic/tumor suppressing functions) were investigated in a panel of PDAC cell lines and liquid biopsies of a small cohort of patients. We used RT-qPCR and droplet digital PCR (ddPCR) to measure the amounts of cellular- and vesicle-associated, and circulating miRNAs. We found that both PDAC cell lines, also after gemcitabine treatment, and patients showed low amounts of cellular-and vesicle-associated miR-155-5p, compared to controls. Interestingly, we did not find any differences when we analyzed circulating miR-155-5p. Furthermore, vesicle-related miR-27a-3p increased in cancer patients compared to the controls, while circulating let-7a-5p, miR-221-3p, miR-23b-3p and miR-193a-3p presented as dysregulated in patients compared to healthy individuals. Our results highlight the potential clinical significance of these analyzed miRNAs as non-invasive diagnostic molecular tools to characterize PDAC.

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Lasp1 Expression Is Implicated in Embryonic Development of Zebrafish

Cited by 5 publications

References 34 publications

Downregulation of Zebrafish Cytosolic Sialidase Neu3.2 Affects Skeletal Muscle Development

Downregulation of Zebrafish Cytosolic Sialidase Neu3.2 Affects Skeletal Muscle Development

Evaluation of DNA methylation levels of SEPT9 and SHOX2 in plasma of patients with head and neck squamous cell carcinoma using droplet digital PCR

Dysregulation of a Subset of Circulating and Vesicle-Associated miRNA in Pancreatic Cancer

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