Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish

Lepanto, Paola - Davison, Camila - Casanova, Gabriela - Badano, José L. - Zolessi, Flavio R.

Resumen:

Background: Retinal ganglion cell (RGC) differentiation in vivo is a highly stereotyped process, likely resulting from the interaction of cell type-specific transcription factors and tissue-derived signaling factors. The primary cilium, as a signaling hub in the cell, may have a role during this process but its presence and localization during RGC generation, and its contribution to the process of cell differentiation, have not been previously assessed in vivo. Methods: In this work we analyzed the distribution of primary cilia in vivo using laser scanning confocal microscopy, as well as their main ultrastructural features by transmission electron microscopy, in the early stages of retinal histogenesis in the zebrafish, around the time of RGC generation and initial differentiation. In addition, we knocked-down ift88 and elipsa, two genes with an essential role in cilia generation and maintenance, a treatment that caused a general reduction in organelle size. The effect on retinal development and RGC differentiation was assessed by confocal microscopy of transgenic or immunolabeled embryos. Results: Our results show that retinal neuroepithelial cells have an apically-localized primary cilium usually protruding from the apical membrane. We also found a small proportion of sub-apical cilia, before and during the neurogenic period. This organelle was also present in an apical position in neuroblasts during apical process retraction and dendritogenesis, although between these stages cilia appeared highly dynamic regarding both presence and position. Disruption of cilia caused a decrease in the proliferation of retinal progenitors and a reduction of neural retina volume. In addition, retinal histogenesis was globally delayed albeit RGC layer formation was preferentially reduced with respect to the amacrine and photoreceptor cell layers. Conclusions: These results indicate that primary cilia exhibit a highly dynamic behavior during early retinal differentiation, and that they are required for the proliferation and survival of retinal progenitors, as well as for neuronal generation, particularly of RGCs.


Detalles Bibliográficos
2016
Cilia
Neurogenesis
Retina
Retinal ganglion cell
Inglés
Universidad de la República
COLIBRI
https://hdl.handle.net/20.500.12008/22083
Acceso abierto
Licencia Creative Commons Atribución (CC –BY 4.0)
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author Lepanto, Paola
author2 Davison, Camila
Casanova, Gabriela
Badano, José L.
Zolessi, Flavio R.
author2_role author
author
author
author
author_facet Lepanto, Paola
Davison, Camila
Casanova, Gabriela
Badano, José L.
Zolessi, Flavio R.
author_role author
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dc.contributor.filiacion.es.fl_str_mv Lepanto, Paola. Instituto Pasteur (Montevideo)
Davison, Camila. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología
Casanova, Gabriela. Universidad de la República (Uruguay). Facultad de Ciencias. Unidad de Microscopía Electrónica
Badano, José L. Instituto Pasteur (Montevideo)
Zolessi, Flavio R. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología
dc.creator.none.fl_str_mv Lepanto, Paola
Davison, Camila
Casanova, Gabriela
Badano, José L.
Zolessi, Flavio R.
dc.date.accessioned.none.fl_str_mv 2019-10-02T22:14:47Z
dc.date.available.none.fl_str_mv 2019-10-02T22:14:47Z
dc.date.issued.es.fl_str_mv 2016
dc.date.submitted.es.fl_str_mv 20191001
dc.description.abstract.none.fl_txt_mv Background: Retinal ganglion cell (RGC) differentiation in vivo is a highly stereotyped process, likely resulting from the interaction of cell type-specific transcription factors and tissue-derived signaling factors. The primary cilium, as a signaling hub in the cell, may have a role during this process but its presence and localization during RGC generation, and its contribution to the process of cell differentiation, have not been previously assessed in vivo. Methods: In this work we analyzed the distribution of primary cilia in vivo using laser scanning confocal microscopy, as well as their main ultrastructural features by transmission electron microscopy, in the early stages of retinal histogenesis in the zebrafish, around the time of RGC generation and initial differentiation. In addition, we knocked-down ift88 and elipsa, two genes with an essential role in cilia generation and maintenance, a treatment that caused a general reduction in organelle size. The effect on retinal development and RGC differentiation was assessed by confocal microscopy of transgenic or immunolabeled embryos. Results: Our results show that retinal neuroepithelial cells have an apically-localized primary cilium usually protruding from the apical membrane. We also found a small proportion of sub-apical cilia, before and during the neurogenic period. This organelle was also present in an apical position in neuroblasts during apical process retraction and dendritogenesis, although between these stages cilia appeared highly dynamic regarding both presence and position. Disruption of cilia caused a decrease in the proliferation of retinal progenitors and a reduction of neural retina volume. In addition, retinal histogenesis was globally delayed albeit RGC layer formation was preferentially reduced with respect to the amacrine and photoreceptor cell layers. Conclusions: These results indicate that primary cilia exhibit a highly dynamic behavior during early retinal differentiation, and that they are required for the proliferation and survival of retinal progenitors, as well as for neuronal generation, particularly of RGCs.
dc.format.mimetype.es.fl_str_mv application/pdf
dc.identifier.citation.es.fl_str_mv Lepanto, P., et al.Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish. Neural Development, 2016, 11 (1), art. no. 10. doi: 10.1186/s13064-016-0064-z
dc.identifier.doi.es.fl_str_mv 10.1186/s13064-016-0064-z
dc.identifier.issn.es.fl_str_mv 1749-8104
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12008/22083
dc.language.iso.none.fl_str_mv en
eng
dc.publisher.es.fl_str_mv BioMed Central Ltd.
dc.relation.ispartof.es.fl_str_mv Neural Development, 2016, 11 (1), art. no. 10
dc.rights.license.none.fl_str_mv Licencia Creative Commons Atribución (CC –BY 4.0)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.source.none.fl_str_mv reponame:COLIBRI
instname:Universidad de la República
instacron:Universidad de la República
dc.subject.es.fl_str_mv Cilia
Neurogenesis
Retina
Retinal ganglion cell
dc.title.none.fl_str_mv Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
dc.type.es.fl_str_mv Artículo
dc.type.none.fl_str_mv info:eu-repo/semantics/article
dc.type.version.none.fl_str_mv info:eu-repo/semantics/publishedVersion
description Background: Retinal ganglion cell (RGC) differentiation in vivo is a highly stereotyped process, likely resulting from the interaction of cell type-specific transcription factors and tissue-derived signaling factors. The primary cilium, as a signaling hub in the cell, may have a role during this process but its presence and localization during RGC generation, and its contribution to the process of cell differentiation, have not been previously assessed in vivo. Methods: In this work we analyzed the distribution of primary cilia in vivo using laser scanning confocal microscopy, as well as their main ultrastructural features by transmission electron microscopy, in the early stages of retinal histogenesis in the zebrafish, around the time of RGC generation and initial differentiation. In addition, we knocked-down ift88 and elipsa, two genes with an essential role in cilia generation and maintenance, a treatment that caused a general reduction in organelle size. The effect on retinal development and RGC differentiation was assessed by confocal microscopy of transgenic or immunolabeled embryos. Results: Our results show that retinal neuroepithelial cells have an apically-localized primary cilium usually protruding from the apical membrane. We also found a small proportion of sub-apical cilia, before and during the neurogenic period. This organelle was also present in an apical position in neuroblasts during apical process retraction and dendritogenesis, although between these stages cilia appeared highly dynamic regarding both presence and position. Disruption of cilia caused a decrease in the proliferation of retinal progenitors and a reduction of neural retina volume. In addition, retinal histogenesis was globally delayed albeit RGC layer formation was preferentially reduced with respect to the amacrine and photoreceptor cell layers. Conclusions: These results indicate that primary cilia exhibit a highly dynamic behavior during early retinal differentiation, and that they are required for the proliferation and survival of retinal progenitors, as well as for neuronal generation, particularly of RGCs.
eu_rights_str_mv openAccess
format article
id COLIBRI_72c499421d3167025bfabc4751fab33a
identifier_str_mv Lepanto, P., et al.Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish. Neural Development, 2016, 11 (1), art. no. 10. doi: 10.1186/s13064-016-0064-z
1749-8104
10.1186/s13064-016-0064-z
instacron_str Universidad de la República
institution Universidad de la República
instname_str Universidad de la República
language eng
language_invalid_str_mv en
network_acronym_str COLIBRI
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oai_identifier_str oai:colibri.udelar.edu.uy:20.500.12008/22083
publishDate 2016
reponame_str COLIBRI
repository.mail.fl_str_mv mabel.seroubian@seciu.edu.uy
repository.name.fl_str_mv COLIBRI - Universidad de la República
repository_id_str 4771
rights_invalid_str_mv Licencia Creative Commons Atribución (CC –BY 4.0)
spelling Lepanto, Paola. Instituto Pasteur (Montevideo)Davison, Camila. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de BiologíaCasanova, Gabriela. Universidad de la República (Uruguay). Facultad de Ciencias. Unidad de Microscopía ElectrónicaBadano, José L. Instituto Pasteur (Montevideo)Zolessi, Flavio R. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología2019-10-02T22:14:47Z2019-10-02T22:14:47Z201620191001Lepanto, P., et al.Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish. Neural Development, 2016, 11 (1), art. no. 10. doi: 10.1186/s13064-016-0064-z1749-8104https://hdl.handle.net/20.500.12008/2208310.1186/s13064-016-0064-zBackground: Retinal ganglion cell (RGC) differentiation in vivo is a highly stereotyped process, likely resulting from the interaction of cell type-specific transcription factors and tissue-derived signaling factors. The primary cilium, as a signaling hub in the cell, may have a role during this process but its presence and localization during RGC generation, and its contribution to the process of cell differentiation, have not been previously assessed in vivo. Methods: In this work we analyzed the distribution of primary cilia in vivo using laser scanning confocal microscopy, as well as their main ultrastructural features by transmission electron microscopy, in the early stages of retinal histogenesis in the zebrafish, around the time of RGC generation and initial differentiation. In addition, we knocked-down ift88 and elipsa, two genes with an essential role in cilia generation and maintenance, a treatment that caused a general reduction in organelle size. The effect on retinal development and RGC differentiation was assessed by confocal microscopy of transgenic or immunolabeled embryos. Results: Our results show that retinal neuroepithelial cells have an apically-localized primary cilium usually protruding from the apical membrane. We also found a small proportion of sub-apical cilia, before and during the neurogenic period. This organelle was also present in an apical position in neuroblasts during apical process retraction and dendritogenesis, although between these stages cilia appeared highly dynamic regarding both presence and position. Disruption of cilia caused a decrease in the proliferation of retinal progenitors and a reduction of neural retina volume. In addition, retinal histogenesis was globally delayed albeit RGC layer formation was preferentially reduced with respect to the amacrine and photoreceptor cell layers. Conclusions: These results indicate that primary cilia exhibit a highly dynamic behavior during early retinal differentiation, and that they are required for the proliferation and survival of retinal progenitors, as well as for neuronal generation, particularly of RGCs.Made available in DSpace on 2019-10-02T22:14:47Z (GMT). No. of bitstreams: 5 101186s130640160064z.pdf: 11152695 bytes, checksum: b3adf1305e7e0cc4c819356bcc97efc3 (MD5) license_text: 38297 bytes, checksum: 4fe6ac477f5a2df0424a5ff1a9bf000c (MD5) license_url: 44 bytes, checksum: a0ebbeafb9d2ec7cbb19d7137ebc392c (MD5) license_rdf: 8067 bytes, checksum: bc1bc9659a4a06e9516479a5adfd8b0e (MD5) license.txt: 4194 bytes, checksum: 7f2e2c17ef6585de66da58d1bfa8b5e1 (MD5) Previous issue date: 2016application/pdfenengBioMed Central Ltd.Neural Development, 2016, 11 (1), art. no. 10Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad De La República. (Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. 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- Universidad de la Repúblicafalse
spellingShingle Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
Lepanto, Paola
Cilia
Neurogenesis
Retina
Retinal ganglion cell
status_str publishedVersion
title Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
title_full Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
title_fullStr Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
title_full_unstemmed Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
title_short Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
title_sort Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish
topic Cilia
Neurogenesis
Retina
Retinal ganglion cell
url https://hdl.handle.net/20.500.12008/22083