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PMID:19123687

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Citation

Hsieh, CS, Ko, CY, Chen, SY, Liu, TM, Wu, JS, Hu, CH and Sun, CK In vivo long-term continuous observation of gene expression in zebrafish embryo nerve systems by using harmonic generation microscopy and morphant technology. J Biomed Opt 13:064041

Abstract

Gene expression plays an important role in embryo development and organ function. Previous studies have shown that harmonic generation microscopy (HGM) can be used as a fluorescence signal-independent, minimally invasive method with a subcellular 3-D resolution and a penetration depth in the order of millimeters for long-term continuous imaging of vertebrate embryos. We show that it is ideal to combine in vivo HGM with the morphant technology for minimally invasive, long-term continuous observation of gene expression in the nervous system of vertebrate embryos. Since second- and third-harmonic generations (SHG, THG) are virtual-state-transition-based systems that depend only on the structure of the organisms, they are not temporally limited by the expression of the fluorescence proteins. We successfully identified the expression of the zarnt2a and the hif-1alpha, 2alpha, and 3alpha genes in the nervous system of zebrafish embryos with specific knockdown genes by microscopically observing the embryos from the early stages of embryogenesis. The results from a combination of the two different modalities, i.e., SHG microscopy and THG microscopy, successfully revealed the weak cell adhesion, cell apoptosis, nerve formation reduction, and neural tube distortion in the morphant zebrafish embryos.

Links

PubMed Online version:10.1117/1.3050423

Keywords

Animals; Animals, Genetically Modified/metabolism; Biotechnology/methods; Brain/embryology; Brain/metabolism; Gene Expression Profiling/methods; Gene Expression Regulation, Developmental/physiology; Microscopy, Confocal/methods; Microscopy, Fluorescence/methods; Zebrafish/embryology; Zebrafish/genetics; Zebrafish/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DANRE:A0A0R4IB28

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:A0A0R4IBR4

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:A0A0R4IRI4

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:A0A2R8QI31

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:A0A2R8QM70

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:A9JRN3

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:B3DJD1

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:F1A7S1

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:F1A7S2

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:F1A7S3

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:Q2EJT3

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:Q6EGR9

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:Q6EHI4

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:Q6PI33

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:Q7T2E4

involved_in

GO:0051402: neuron apoptotic process

ECO:0000316: genetic interaction evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-7,ZFIN:ZDB-MRPHLNO-100414-8,ZFIN:ZDB-MRPHLNO-100610-11

P

Seeded From UniProt

complete

DANRE:ARNT2

involved_in

GO:0098609: cell-cell adhesion

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-100414-6

P

Seeded From UniProt

complete


See also

References

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