Development of interspecies testicular germ-cell transplantation in flatfish
Interspecific testicular germ cell (TGC) transplantation was investigated in two commercial flatfish species. Testes from donor species (Senegalese sole) were evaluated using classical histological techniques (haematoxylin-eosin staining and haematoxylin-light green-orange G-acid fuchsine staining),...
Published in: | Reproduction, Fertility and Development |
---|---|
Main Authors: | , , |
Format: | Article in Journal/Newspaper |
Language: | unknown |
Published: |
CSIRO Publishing
2014
|
Subjects: | |
Online Access: | http://hdl.handle.net/10261/102072 https://doi.org/10.1071/RD13103 |
id |
ftcsic:oai:digital.csic.es:10261/102072 |
---|---|
record_format |
openpolar |
spelling |
ftcsic:oai:digital.csic.es:10261/102072 2024-02-11T10:09:16+01:00 Development of interspecies testicular germ-cell transplantation in flatfish Pacchiarini, Tiziana Sarasquete, Carmen Cabrita, Elsa 2014-06-05 http://hdl.handle.net/10261/102072 https://doi.org/10.1071/RD13103 unknown CSIRO Publishing http://doi.org/10.1071/RD13103 doi:10.1071/RD13103 issn: 1031-3613 e-issn: 1448-5990 Reproduction, Fertility and Development 26(5): 690-702 (2014) http://hdl.handle.net/10261/102072 none Turbot Spermatogonia Vasa Xenotransplantation Senegalese sole Microinjection artículo http://purl.org/coar/resource_type/c_6501 2014 ftcsic https://doi.org/10.1071/RD13103 2024-01-16T10:01:11Z Interspecific testicular germ cell (TGC) transplantation was investigated in two commercial flatfish species. Testes from donor species (Senegalese sole) were evaluated using classical histological techniques (haematoxylin-eosin staining and haematoxylin-light green-orange G-acid fuchsine staining), in situ hybridisation and immunohistochemical analysis. Both Ssvasa1-2 mRNAs and SsVasa protein allowed the characterisation of TGCs, confirming the usefulness of the vasa gene in the detection of Senegalese sole TGCs. Xenogenic transplants were carried out using TGCs from oneyear- old Senegalese sole into turbot larvae. Propidium iodide-SYBR-14 and 40,60-diamidino-2-phenylindole (DAPI) staining showed that 87.98% of the extracted testicular cells were viable for microinjection and that 15.63% of the total recovered cells were spermatogonia. The vasa gene was characterised in turbot recipients using cDNA cloning. Smvasa mRNA was confirmed as a germ cell-specific molecular marker in this species. Smvasa expression analysis during turbot ontogeny was carried out before Senegalese soleTGCtransplants into turbot larvae. Turbot larvae at 18 days after hatching (DAH) proved to be susceptible to manipulation procedures. High survival rates (83.75±15.90-100%) were obtained for turbot larvae at 27, 34 and 42 DAH. These data highlight the huge potential of this species for transplantation studies. Quantitative PCR was employed to detect Senegalese sole vasa mRNAs (Ssvasa1-2) in the recipient turbot larvae. The Ssvasa mRNAs showed a significant increase in relative expression in 42-DAH microinjected larvae three weeks after treatment, showing the proliferation of Senegalese sole spermatogonia in transplanted turbot larvae. © CSIRO 2014. Peer Reviewed Article in Journal/Newspaper Turbot Digital.CSIC (Spanish National Research Council) Vasa ENVELOPE(25.177,25.177,67.587,67.587) Reproduction, Fertility and Development 26 5 690 |
institution |
Open Polar |
collection |
Digital.CSIC (Spanish National Research Council) |
op_collection_id |
ftcsic |
language |
unknown |
topic |
Turbot Spermatogonia Vasa Xenotransplantation Senegalese sole Microinjection |
spellingShingle |
Turbot Spermatogonia Vasa Xenotransplantation Senegalese sole Microinjection Pacchiarini, Tiziana Sarasquete, Carmen Cabrita, Elsa Development of interspecies testicular germ-cell transplantation in flatfish |
topic_facet |
Turbot Spermatogonia Vasa Xenotransplantation Senegalese sole Microinjection |
description |
Interspecific testicular germ cell (TGC) transplantation was investigated in two commercial flatfish species. Testes from donor species (Senegalese sole) were evaluated using classical histological techniques (haematoxylin-eosin staining and haematoxylin-light green-orange G-acid fuchsine staining), in situ hybridisation and immunohistochemical analysis. Both Ssvasa1-2 mRNAs and SsVasa protein allowed the characterisation of TGCs, confirming the usefulness of the vasa gene in the detection of Senegalese sole TGCs. Xenogenic transplants were carried out using TGCs from oneyear- old Senegalese sole into turbot larvae. Propidium iodide-SYBR-14 and 40,60-diamidino-2-phenylindole (DAPI) staining showed that 87.98% of the extracted testicular cells were viable for microinjection and that 15.63% of the total recovered cells were spermatogonia. The vasa gene was characterised in turbot recipients using cDNA cloning. Smvasa mRNA was confirmed as a germ cell-specific molecular marker in this species. Smvasa expression analysis during turbot ontogeny was carried out before Senegalese soleTGCtransplants into turbot larvae. Turbot larvae at 18 days after hatching (DAH) proved to be susceptible to manipulation procedures. High survival rates (83.75±15.90-100%) were obtained for turbot larvae at 27, 34 and 42 DAH. These data highlight the huge potential of this species for transplantation studies. Quantitative PCR was employed to detect Senegalese sole vasa mRNAs (Ssvasa1-2) in the recipient turbot larvae. The Ssvasa mRNAs showed a significant increase in relative expression in 42-DAH microinjected larvae three weeks after treatment, showing the proliferation of Senegalese sole spermatogonia in transplanted turbot larvae. © CSIRO 2014. Peer Reviewed |
format |
Article in Journal/Newspaper |
author |
Pacchiarini, Tiziana Sarasquete, Carmen Cabrita, Elsa |
author_facet |
Pacchiarini, Tiziana Sarasquete, Carmen Cabrita, Elsa |
author_sort |
Pacchiarini, Tiziana |
title |
Development of interspecies testicular germ-cell transplantation in flatfish |
title_short |
Development of interspecies testicular germ-cell transplantation in flatfish |
title_full |
Development of interspecies testicular germ-cell transplantation in flatfish |
title_fullStr |
Development of interspecies testicular germ-cell transplantation in flatfish |
title_full_unstemmed |
Development of interspecies testicular germ-cell transplantation in flatfish |
title_sort |
development of interspecies testicular germ-cell transplantation in flatfish |
publisher |
CSIRO Publishing |
publishDate |
2014 |
url |
http://hdl.handle.net/10261/102072 https://doi.org/10.1071/RD13103 |
long_lat |
ENVELOPE(25.177,25.177,67.587,67.587) |
geographic |
Vasa |
geographic_facet |
Vasa |
genre |
Turbot |
genre_facet |
Turbot |
op_relation |
http://doi.org/10.1071/RD13103 doi:10.1071/RD13103 issn: 1031-3613 e-issn: 1448-5990 Reproduction, Fertility and Development 26(5): 690-702 (2014) http://hdl.handle.net/10261/102072 |
op_rights |
none |
op_doi |
https://doi.org/10.1071/RD13103 |
container_title |
Reproduction, Fertility and Development |
container_volume |
26 |
container_issue |
5 |
container_start_page |
690 |
_version_ |
1790609058658516992 |