Validation of commonly used reference genes for sleep-related gene expression studies
Autor(a) principal: | |
---|---|
Data de Publicação: | 2009 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/31521 http://dx.doi.org/10.1186/1471-2199-10-45 |
Resumo: | Background: Sleep is a restorative process and is essential for maintenance of mental and physical health. in an attempt to understand the complexity of sleep, multidisciplinary strategies, including genetic approaches, have been applied to sleep research. Although quantitative real time PCR has been used in previous sleep-related gene expression studies, proper validation of reference genes is currently lacking. Thus, we examined the effect of total or paradoxical sleep deprivation (TSD or PSD) on the expression stability of the following frequently used reference genes in brain and blood: beta-actin (b-actin), beta-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and hypoxanthine guanine phosphoribosyl transferase (HPRT).Results: Neither TSD nor PSD affected the expression stability of all tested genes in both tissues indicating that b-actin, B2M, GAPDH and HPRT are appropriate reference genes for the sleep-related gene expression studies. in order to further verify these results, the relative expression of brain derived neurotrophic factor (BDNF) and glycerol-3-phosphate dehydrogenase1 (GPD1) was evaluated in brain and blood, respectively. the normalization with each of four reference genes produced similar pattern of expression in control and sleep deprived rats, but subtle differences in the magnitude of expression fold change were observed which might affect the statistical significance.Conclusion: This study demonstrated that sleep deprivation does not alter the expression stability of commonly used reference genes in brain and blood. Nonetheless, the use of multiple reference genes in quantitative RT-PCR is required for the accurate results. |
id |
UFSP_16281e3401da57a0b6f6f6daf2289b3d |
---|---|
oai_identifier_str |
oai:repositorio.unifesp.br:11600/31521 |
network_acronym_str |
UFSP |
network_name_str |
Repositório Institucional da UNIFESP |
repository_id_str |
3465 |
spelling |
Lee, Kil Sun [UNIFESP]Alvarenga, Tathiana Aparecida [UNIFESP]Guindalini, Camila [UNIFESP]Andersen, Monica Levy [UNIFESP]Castro, Rosa Maria Rodrigues Pinto Santos [UNIFESP]Tufik, Sergio [UNIFESP]AFIPUniversidade Federal de São Paulo (UNIFESP)2016-01-24T13:52:33Z2016-01-24T13:52:33Z2009-05-15Bmc Molecular Biology. London: Biomed Central Ltd, v. 10, 8 p., 2009.1471-2199http://repositorio.unifesp.br/handle/11600/31521http://dx.doi.org/10.1186/1471-2199-10-45WOS000266807500001.pdf10.1186/1471-2199-10-45WOS:000266807500001Background: Sleep is a restorative process and is essential for maintenance of mental and physical health. in an attempt to understand the complexity of sleep, multidisciplinary strategies, including genetic approaches, have been applied to sleep research. Although quantitative real time PCR has been used in previous sleep-related gene expression studies, proper validation of reference genes is currently lacking. Thus, we examined the effect of total or paradoxical sleep deprivation (TSD or PSD) on the expression stability of the following frequently used reference genes in brain and blood: beta-actin (b-actin), beta-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and hypoxanthine guanine phosphoribosyl transferase (HPRT).Results: Neither TSD nor PSD affected the expression stability of all tested genes in both tissues indicating that b-actin, B2M, GAPDH and HPRT are appropriate reference genes for the sleep-related gene expression studies. in order to further verify these results, the relative expression of brain derived neurotrophic factor (BDNF) and glycerol-3-phosphate dehydrogenase1 (GPD1) was evaluated in brain and blood, respectively. the normalization with each of four reference genes produced similar pattern of expression in control and sleep deprived rats, but subtle differences in the magnitude of expression fold change were observed which might affect the statistical significance.Conclusion: This study demonstrated that sleep deprivation does not alter the expression stability of commonly used reference genes in brain and blood. Nonetheless, the use of multiple reference genes in quantitative RT-PCR is required for the accurate results.Associacao Fundo de Incentivo a Psicofarmacologia (AFIP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)AFIPAFIP, BR-04024002 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, BR-04024002 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, BR-04024002 São Paulo, BrazilFAPESP: 98/14303-3FAPESP: 06/58274-5Web of Science8engBiomed Central LtdBmc Molecular BiologyValidation of commonly used reference genes for sleep-related gene expression studiesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000266807500001.pdfapplication/pdf337507${dspace.ui.url}/bitstream/11600/31521/1/WOS000266807500001.pdf278b62d3c73337e72914265c6ef5c44cMD51open accessTEXTWOS000266807500001.pdf.txtWOS000266807500001.pdf.txtExtracted texttext/plain33374${dspace.ui.url}/bitstream/11600/31521/2/WOS000266807500001.pdf.txt18dc0987d686336f528f344038f1fb80MD52open access11600/315212022-06-02 09:35:06.162open accessoai:repositorio.unifesp.br:11600/31521Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:16:22.269733Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Validation of commonly used reference genes for sleep-related gene expression studies |
title |
Validation of commonly used reference genes for sleep-related gene expression studies |
spellingShingle |
Validation of commonly used reference genes for sleep-related gene expression studies Lee, Kil Sun [UNIFESP] |
title_short |
Validation of commonly used reference genes for sleep-related gene expression studies |
title_full |
Validation of commonly used reference genes for sleep-related gene expression studies |
title_fullStr |
Validation of commonly used reference genes for sleep-related gene expression studies |
title_full_unstemmed |
Validation of commonly used reference genes for sleep-related gene expression studies |
title_sort |
Validation of commonly used reference genes for sleep-related gene expression studies |
author |
Lee, Kil Sun [UNIFESP] |
author_facet |
Lee, Kil Sun [UNIFESP] Alvarenga, Tathiana Aparecida [UNIFESP] Guindalini, Camila [UNIFESP] Andersen, Monica Levy [UNIFESP] Castro, Rosa Maria Rodrigues Pinto Santos [UNIFESP] Tufik, Sergio [UNIFESP] |
author_role |
author |
author2 |
Alvarenga, Tathiana Aparecida [UNIFESP] Guindalini, Camila [UNIFESP] Andersen, Monica Levy [UNIFESP] Castro, Rosa Maria Rodrigues Pinto Santos [UNIFESP] Tufik, Sergio [UNIFESP] |
author2_role |
author author author author author |
dc.contributor.institution.none.fl_str_mv |
AFIP Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Lee, Kil Sun [UNIFESP] Alvarenga, Tathiana Aparecida [UNIFESP] Guindalini, Camila [UNIFESP] Andersen, Monica Levy [UNIFESP] Castro, Rosa Maria Rodrigues Pinto Santos [UNIFESP] Tufik, Sergio [UNIFESP] |
description |
Background: Sleep is a restorative process and is essential for maintenance of mental and physical health. in an attempt to understand the complexity of sleep, multidisciplinary strategies, including genetic approaches, have been applied to sleep research. Although quantitative real time PCR has been used in previous sleep-related gene expression studies, proper validation of reference genes is currently lacking. Thus, we examined the effect of total or paradoxical sleep deprivation (TSD or PSD) on the expression stability of the following frequently used reference genes in brain and blood: beta-actin (b-actin), beta-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and hypoxanthine guanine phosphoribosyl transferase (HPRT).Results: Neither TSD nor PSD affected the expression stability of all tested genes in both tissues indicating that b-actin, B2M, GAPDH and HPRT are appropriate reference genes for the sleep-related gene expression studies. in order to further verify these results, the relative expression of brain derived neurotrophic factor (BDNF) and glycerol-3-phosphate dehydrogenase1 (GPD1) was evaluated in brain and blood, respectively. the normalization with each of four reference genes produced similar pattern of expression in control and sleep deprived rats, but subtle differences in the magnitude of expression fold change were observed which might affect the statistical significance.Conclusion: This study demonstrated that sleep deprivation does not alter the expression stability of commonly used reference genes in brain and blood. Nonetheless, the use of multiple reference genes in quantitative RT-PCR is required for the accurate results. |
publishDate |
2009 |
dc.date.issued.fl_str_mv |
2009-05-15 |
dc.date.accessioned.fl_str_mv |
2016-01-24T13:52:33Z |
dc.date.available.fl_str_mv |
2016-01-24T13:52:33Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Bmc Molecular Biology. London: Biomed Central Ltd, v. 10, 8 p., 2009. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/31521 http://dx.doi.org/10.1186/1471-2199-10-45 |
dc.identifier.issn.none.fl_str_mv |
1471-2199 |
dc.identifier.file.none.fl_str_mv |
WOS000266807500001.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1186/1471-2199-10-45 |
dc.identifier.wos.none.fl_str_mv |
WOS:000266807500001 |
identifier_str_mv |
Bmc Molecular Biology. London: Biomed Central Ltd, v. 10, 8 p., 2009. 1471-2199 WOS000266807500001.pdf 10.1186/1471-2199-10-45 WOS:000266807500001 |
url |
http://repositorio.unifesp.br/handle/11600/31521 http://dx.doi.org/10.1186/1471-2199-10-45 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Bmc Molecular Biology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
8 |
dc.publisher.none.fl_str_mv |
Biomed Central Ltd |
publisher.none.fl_str_mv |
Biomed Central Ltd |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
bitstream.url.fl_str_mv |
${dspace.ui.url}/bitstream/11600/31521/1/WOS000266807500001.pdf ${dspace.ui.url}/bitstream/11600/31521/2/WOS000266807500001.pdf.txt |
bitstream.checksum.fl_str_mv |
278b62d3c73337e72914265c6ef5c44c 18dc0987d686336f528f344038f1fb80 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
_version_ |
1783460270281064448 |