Cargando…
In Vitro and In Vivo Determinations of The Anti-GDNF Family Receptor Alpha 1 Antibody in Mice by Immunochemistry and RT-PCR
BACKGROUND: The glial cell-derived neurotrophic factor (GDNF) family plays essential roles in the maintenance, growth, regulatory and signalling pathways of spermatogonial stem cells (SSCs). In this study, we analysed the ex- pression of anti-GDNF family receptor alpha 1 antibody (GFRa1) by immunohi...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royan Institute
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7604702/ https://www.ncbi.nlm.nih.gov/pubmed/33098391 http://dx.doi.org/10.22074/ijfs.2020.6051 |
_version_ | 1783604187413610496 |
---|---|
author | Azizi, Hossein Niazi Tabar, Amirreza Skutella, Thomas Govahi, Mostafa |
author_facet | Azizi, Hossein Niazi Tabar, Amirreza Skutella, Thomas Govahi, Mostafa |
author_sort | Azizi, Hossein |
collection | PubMed |
description | BACKGROUND: The glial cell-derived neurotrophic factor (GDNF) family plays essential roles in the maintenance, growth, regulatory and signalling pathways of spermatogonial stem cells (SSCs). In this study, we analysed the ex- pression of anti-GDNF family receptor alpha 1 antibody (GFRa1) by immunohistochemistry (IHC), immunocyto- chemistry (ICC), Fluidigm real-time polymerase chain reaction (RT-PCR) and flow cytometry analyses. MATERIALS AND METHODS: In this experiment study, ICC, IHC, Fluidigm RT-PCR and flow cytometry were used to analyse the expression of the germ cell marker GFRa1 in testis tissue and SSC culture. RESULTS: IHC analysis showed that there were two groups of GFRa1 positive cells in the seminiferous tubules based on their location and expression shape - a small round punctuated shape on the basal compartment donut shape and a C-shaped expression located between the basal and the luminal compartments of the seminiferous tubules. OCT4 and PLZF positive cells may have similar patterns of expression as the first group. Assessment of the seminiferous tubule sections demonstrated that about 27% of the SSCs were positive for GFRa1. Fluidigm RT-PCR confirmed the significant expression (P<0.001) of GFRa1 in the SSCs compared to testicular stromal cells (TSCs). Flow cytometry analysis demonstrated that about 75% of the isolated SSCs colonies were positive for GFRa1. CONCLUSION: The results indicated that GFRa1 had a specific expression pattern both in vivo and in vitro. This finding could be helpful for understanding the proliferation, maintenance and signalling pathways of SSCs, and differentiation of meiotic and haploid germ cells. |
format | Online Article Text |
id | pubmed-7604702 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Royan Institute |
record_format | MEDLINE/PubMed |
spelling | pubmed-76047022020-11-06 In Vitro and In Vivo Determinations of The Anti-GDNF Family Receptor Alpha 1 Antibody in Mice by Immunochemistry and RT-PCR Azizi, Hossein Niazi Tabar, Amirreza Skutella, Thomas Govahi, Mostafa Int J Fertil Steril Original Article BACKGROUND: The glial cell-derived neurotrophic factor (GDNF) family plays essential roles in the maintenance, growth, regulatory and signalling pathways of spermatogonial stem cells (SSCs). In this study, we analysed the ex- pression of anti-GDNF family receptor alpha 1 antibody (GFRa1) by immunohistochemistry (IHC), immunocyto- chemistry (ICC), Fluidigm real-time polymerase chain reaction (RT-PCR) and flow cytometry analyses. MATERIALS AND METHODS: In this experiment study, ICC, IHC, Fluidigm RT-PCR and flow cytometry were used to analyse the expression of the germ cell marker GFRa1 in testis tissue and SSC culture. RESULTS: IHC analysis showed that there were two groups of GFRa1 positive cells in the seminiferous tubules based on their location and expression shape - a small round punctuated shape on the basal compartment donut shape and a C-shaped expression located between the basal and the luminal compartments of the seminiferous tubules. OCT4 and PLZF positive cells may have similar patterns of expression as the first group. Assessment of the seminiferous tubule sections demonstrated that about 27% of the SSCs were positive for GFRa1. Fluidigm RT-PCR confirmed the significant expression (P<0.001) of GFRa1 in the SSCs compared to testicular stromal cells (TSCs). Flow cytometry analysis demonstrated that about 75% of the isolated SSCs colonies were positive for GFRa1. CONCLUSION: The results indicated that GFRa1 had a specific expression pattern both in vivo and in vitro. This finding could be helpful for understanding the proliferation, maintenance and signalling pathways of SSCs, and differentiation of meiotic and haploid germ cells. Royan Institute 2020 2020-10-12 /pmc/articles/PMC7604702/ /pubmed/33098391 http://dx.doi.org/10.22074/ijfs.2020.6051 Text en Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Azizi, Hossein Niazi Tabar, Amirreza Skutella, Thomas Govahi, Mostafa In Vitro and In Vivo Determinations of The Anti-GDNF Family Receptor Alpha 1 Antibody in Mice by Immunochemistry and RT-PCR |
title | In Vitro and In Vivo Determinations of The Anti-GDNF
Family Receptor Alpha 1 Antibody in Mice by Immunochemistry
and RT-PCR |
title_full | In Vitro and In Vivo Determinations of The Anti-GDNF
Family Receptor Alpha 1 Antibody in Mice by Immunochemistry
and RT-PCR |
title_fullStr | In Vitro and In Vivo Determinations of The Anti-GDNF
Family Receptor Alpha 1 Antibody in Mice by Immunochemistry
and RT-PCR |
title_full_unstemmed | In Vitro and In Vivo Determinations of The Anti-GDNF
Family Receptor Alpha 1 Antibody in Mice by Immunochemistry
and RT-PCR |
title_short | In Vitro and In Vivo Determinations of The Anti-GDNF
Family Receptor Alpha 1 Antibody in Mice by Immunochemistry
and RT-PCR |
title_sort | in vitro and in vivo determinations of the anti-gdnf
family receptor alpha 1 antibody in mice by immunochemistry
and rt-pcr |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7604702/ https://www.ncbi.nlm.nih.gov/pubmed/33098391 http://dx.doi.org/10.22074/ijfs.2020.6051 |
work_keys_str_mv | AT azizihossein invitroandinvivodeterminationsoftheantigdnffamilyreceptoralpha1antibodyinmicebyimmunochemistryandrtpcr AT niazitabaramirreza invitroandinvivodeterminationsoftheantigdnffamilyreceptoralpha1antibodyinmicebyimmunochemistryandrtpcr AT skutellathomas invitroandinvivodeterminationsoftheantigdnffamilyreceptoralpha1antibodyinmicebyimmunochemistryandrtpcr AT govahimostafa invitroandinvivodeterminationsoftheantigdnffamilyreceptoralpha1antibodyinmicebyimmunochemistryandrtpcr |