Zebrafish: an exciting model for investigating the spatio-temporal pattern of enteric nervous system development.

Hdl Handle:
http://hdl.handle.net/10147/207443
Title:
Zebrafish: an exciting model for investigating the spatio-temporal pattern of enteric nervous system development.
Authors:
Doodnath, Reshma; Dervan, Adrian; Wride, Michael A; Puri, Prem
Affiliation:
National Children's Research Centre, Our Lady's Children's Hospital, Crumlin,, Dublin 12, Ireland. rdoodnath@gmail.com
Citation:
Pediatr Surg Int. 2010 Dec;26(12):1217-21. Epub 2010 Oct 24.
Journal:
Pediatric surgery international
Issue Date:
1-Feb-2012
URI:
http://hdl.handle.net/10147/207443
DOI:
10.1007/s00383-010-2746-7
PubMed ID:
20972797
Abstract:
AIM: Recently, the zebrafish (Danio rerio) has been shown to be an excellent model for human paediatric research. Advantages over other models include its small size, externally visually accessible development and ease of experimental manipulation. The enteric nervous system (ENS) consists of neurons and enteric glia. Glial cells permit cell bodies and processes of neurons to be arranged and maintained in a proper spatial arrangement, and are essential in the maintenance of basic physiological functions of neurons. Glial fibrillary acidic protein (GFAP) is expressed in astrocytes, but also expressed outside of the central nervous system. The aim of this study was to investigate the spatio-temporal pattern of GFAP expression in developing zebrafish ENS from 24 h post-fertilization (hpf), using transgenic fish that express green fluorescent protein (GFP). METHODS: Zebrafish embryos were collected from transgenic GFP Tg(GFAP:GFP)(mi2001) adult zebrafish from 24 to 120 hpf, fixed and processed for whole mount immunohistochemistry. Antibodies to Phox2b were used to identify enteric neurons. Specimens were mounted on slides and imaging was performed using a fluorescent laser confocal microscope. RESULTS: GFAP:GFP labelling outside the spinal cord was identified in embryos from 48 hpf. The patterning was intracellular and consisted of elongated profiles that appeared to migrate away from the spinal cord into the periphery. At 72 and 96 hpf, GFAP:GFP was expressed dorsally and ventrally to the intestinal tract. At 120 hpf, GFAP:GFP was expressed throughout the intestinal wall, and clusters of enteric neurons were identified using Phox2b immunofluorescence along the pathway of GFAP:GFP positive processes, indicative of a migratory pathway of ENS precursors from the spinal cord into the intestine. CONCLUSION: The pattern of migration of GFAP:GFP expressing cells outside the spinal cord suggests an organized, early developing migratory pathway to the ENS. This shows for the first time that Tg(GFAP:GFP)(mi2001) zebrafish model is an ideal one to study spatio-temporal patterning of early ENS development.
Language:
eng
MeSH:
Animals; Animals, Genetically Modified; Cell Movement; Enteric Nervous System/*embryology; Fluorescent Antibody Technique; Glial Fibrillary Acidic Protein/*metabolism; Homeodomain Proteins; Humans; Transcription Factors; *Zebrafish
ISSN:
1437-9813 (Electronic); 0179-0358 (Linking)

Full metadata record

DC FieldValue Language
dc.contributor.authorDoodnath, Reshmaen_GB
dc.contributor.authorDervan, Adrianen_GB
dc.contributor.authorWride, Michael Aen_GB
dc.contributor.authorPuri, Premen_GB
dc.date.accessioned2012-02-01T10:25:08Z-
dc.date.available2012-02-01T10:25:08Z-
dc.date.issued2012-02-01T10:25:08Z-
dc.identifier.citationPediatr Surg Int. 2010 Dec;26(12):1217-21. Epub 2010 Oct 24.en_GB
dc.identifier.issn1437-9813 (Electronic)en_GB
dc.identifier.issn0179-0358 (Linking)en_GB
dc.identifier.pmid20972797en_GB
dc.identifier.doi10.1007/s00383-010-2746-7en_GB
dc.identifier.urihttp://hdl.handle.net/10147/207443-
dc.description.abstractAIM: Recently, the zebrafish (Danio rerio) has been shown to be an excellent model for human paediatric research. Advantages over other models include its small size, externally visually accessible development and ease of experimental manipulation. The enteric nervous system (ENS) consists of neurons and enteric glia. Glial cells permit cell bodies and processes of neurons to be arranged and maintained in a proper spatial arrangement, and are essential in the maintenance of basic physiological functions of neurons. Glial fibrillary acidic protein (GFAP) is expressed in astrocytes, but also expressed outside of the central nervous system. The aim of this study was to investigate the spatio-temporal pattern of GFAP expression in developing zebrafish ENS from 24 h post-fertilization (hpf), using transgenic fish that express green fluorescent protein (GFP). METHODS: Zebrafish embryos were collected from transgenic GFP Tg(GFAP:GFP)(mi2001) adult zebrafish from 24 to 120 hpf, fixed and processed for whole mount immunohistochemistry. Antibodies to Phox2b were used to identify enteric neurons. Specimens were mounted on slides and imaging was performed using a fluorescent laser confocal microscope. RESULTS: GFAP:GFP labelling outside the spinal cord was identified in embryos from 48 hpf. The patterning was intracellular and consisted of elongated profiles that appeared to migrate away from the spinal cord into the periphery. At 72 and 96 hpf, GFAP:GFP was expressed dorsally and ventrally to the intestinal tract. At 120 hpf, GFAP:GFP was expressed throughout the intestinal wall, and clusters of enteric neurons were identified using Phox2b immunofluorescence along the pathway of GFAP:GFP positive processes, indicative of a migratory pathway of ENS precursors from the spinal cord into the intestine. CONCLUSION: The pattern of migration of GFAP:GFP expressing cells outside the spinal cord suggests an organized, early developing migratory pathway to the ENS. This shows for the first time that Tg(GFAP:GFP)(mi2001) zebrafish model is an ideal one to study spatio-temporal patterning of early ENS development.en_GB
dc.language.isoengen_GB
dc.subject.meshAnimalsen_GB
dc.subject.meshAnimals, Genetically Modifieden_GB
dc.subject.meshCell Movementen_GB
dc.subject.meshEnteric Nervous System/*embryologyen_GB
dc.subject.meshFluorescent Antibody Techniqueen_GB
dc.subject.meshGlial Fibrillary Acidic Protein/*metabolismen_GB
dc.subject.meshHomeodomain Proteinsen_GB
dc.subject.meshHumansen_GB
dc.subject.meshTranscription Factorsen_GB
dc.subject.mesh*Zebrafishen_GB
dc.titleZebrafish: an exciting model for investigating the spatio-temporal pattern of enteric nervous system development.en_GB
dc.contributor.departmentNational Children's Research Centre, Our Lady's Children's Hospital, Crumlin,, Dublin 12, Ireland. rdoodnath@gmail.comen_GB
dc.identifier.journalPediatric surgery internationalen_GB
dc.description.provinceLeinster-
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