Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization

Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Cappelli, Katia [verfasserIn] Felicetti, Michela Capomaccio, Stefano Spinsanti, Giacomo Silvestrelli, Maurizio Supplizi, Andrea Verini

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2008

Schlagwörter:	Reference Gene Candidate Reference Gene Stable Reference Gene Hypoxanthine Phosphoribosyltransferase Good Reference Gene

Anmerkung:	© Cappelli et al; licensee BioMed Central Ltd. 2008

Übergeordnetes Werk:	Enthalten in: BMC molecular biology - London : BioMed Central, 2000, 9(2008), 1 vom: 19. Mai
Übergeordnetes Werk:	volume:9 ; year:2008 ; number:1 ; day:19 ; month:05

Links:	Volltext

DOI / URN:	10.1186/1471-2199-9-49

Katalog-ID:	SPR027216411

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245	1	0	\|a Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization
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520			\|a Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool.
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700	1		\|a Supplizi, Andrea Verini \|4 aut
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matchkey_str	article:14712199:2008----::xrienuesrsihreslcinfhmssalrfrneeefrun
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publishDate	2008
allfields	10.1186/1471-2199-9-49 doi (DE-627)SPR027216411 (SPR)1471-2199-9-49-e DE-627 ger DE-627 rakwb eng Cappelli, Katia verfasserin aut Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization 2008 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Cappelli et al; licensee BioMed Central Ltd. 2008 Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. Reference Gene (dpeaa)DE-He213 Candidate Reference Gene (dpeaa)DE-He213 Stable Reference Gene (dpeaa)DE-He213 Hypoxanthine Phosphoribosyltransferase (dpeaa)DE-He213 Good Reference Gene (dpeaa)DE-He213 Felicetti, Michela aut Capomaccio, Stefano aut Spinsanti, Giacomo aut Silvestrelli, Maurizio aut Supplizi, Andrea Verini aut Enthalten in BMC molecular biology London : BioMed Central, 2000 9(2008), 1 vom: 19. Mai (DE-627)326645004 (DE-600)2041506-0 1471-2199 nnns volume:9 year:2008 number:1 day:19 month:05 https://dx.doi.org/10.1186/1471-2199-9-49 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2008 1 19 05
spelling	10.1186/1471-2199-9-49 doi (DE-627)SPR027216411 (SPR)1471-2199-9-49-e DE-627 ger DE-627 rakwb eng Cappelli, Katia verfasserin aut Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization 2008 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Cappelli et al; licensee BioMed Central Ltd. 2008 Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. Reference Gene (dpeaa)DE-He213 Candidate Reference Gene (dpeaa)DE-He213 Stable Reference Gene (dpeaa)DE-He213 Hypoxanthine Phosphoribosyltransferase (dpeaa)DE-He213 Good Reference Gene (dpeaa)DE-He213 Felicetti, Michela aut Capomaccio, Stefano aut Spinsanti, Giacomo aut Silvestrelli, Maurizio aut Supplizi, Andrea Verini aut Enthalten in BMC molecular biology London : BioMed Central, 2000 9(2008), 1 vom: 19. Mai (DE-627)326645004 (DE-600)2041506-0 1471-2199 nnns volume:9 year:2008 number:1 day:19 month:05 https://dx.doi.org/10.1186/1471-2199-9-49 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2008 1 19 05
allfields_unstemmed	10.1186/1471-2199-9-49 doi (DE-627)SPR027216411 (SPR)1471-2199-9-49-e DE-627 ger DE-627 rakwb eng Cappelli, Katia verfasserin aut Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization 2008 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Cappelli et al; licensee BioMed Central Ltd. 2008 Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. Reference Gene (dpeaa)DE-He213 Candidate Reference Gene (dpeaa)DE-He213 Stable Reference Gene (dpeaa)DE-He213 Hypoxanthine Phosphoribosyltransferase (dpeaa)DE-He213 Good Reference Gene (dpeaa)DE-He213 Felicetti, Michela aut Capomaccio, Stefano aut Spinsanti, Giacomo aut Silvestrelli, Maurizio aut Supplizi, Andrea Verini aut Enthalten in BMC molecular biology London : BioMed Central, 2000 9(2008), 1 vom: 19. Mai (DE-627)326645004 (DE-600)2041506-0 1471-2199 nnns volume:9 year:2008 number:1 day:19 month:05 https://dx.doi.org/10.1186/1471-2199-9-49 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2008 1 19 05
allfieldsGer	10.1186/1471-2199-9-49 doi (DE-627)SPR027216411 (SPR)1471-2199-9-49-e DE-627 ger DE-627 rakwb eng Cappelli, Katia verfasserin aut Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization 2008 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Cappelli et al; licensee BioMed Central Ltd. 2008 Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. Reference Gene (dpeaa)DE-He213 Candidate Reference Gene (dpeaa)DE-He213 Stable Reference Gene (dpeaa)DE-He213 Hypoxanthine Phosphoribosyltransferase (dpeaa)DE-He213 Good Reference Gene (dpeaa)DE-He213 Felicetti, Michela aut Capomaccio, Stefano aut Spinsanti, Giacomo aut Silvestrelli, Maurizio aut Supplizi, Andrea Verini aut Enthalten in BMC molecular biology London : BioMed Central, 2000 9(2008), 1 vom: 19. Mai (DE-627)326645004 (DE-600)2041506-0 1471-2199 nnns volume:9 year:2008 number:1 day:19 month:05 https://dx.doi.org/10.1186/1471-2199-9-49 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2008 1 19 05
allfieldsSound	10.1186/1471-2199-9-49 doi (DE-627)SPR027216411 (SPR)1471-2199-9-49-e DE-627 ger DE-627 rakwb eng Cappelli, Katia verfasserin aut Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization 2008 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Cappelli et al; licensee BioMed Central Ltd. 2008 Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. Reference Gene (dpeaa)DE-He213 Candidate Reference Gene (dpeaa)DE-He213 Stable Reference Gene (dpeaa)DE-He213 Hypoxanthine Phosphoribosyltransferase (dpeaa)DE-He213 Good Reference Gene (dpeaa)DE-He213 Felicetti, Michela aut Capomaccio, Stefano aut Spinsanti, Giacomo aut Silvestrelli, Maurizio aut Supplizi, Andrea Verini aut Enthalten in BMC molecular biology London : BioMed Central, 2000 9(2008), 1 vom: 19. Mai (DE-627)326645004 (DE-600)2041506-0 1471-2199 nnns volume:9 year:2008 number:1 day:19 month:05 https://dx.doi.org/10.1186/1471-2199-9-49 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2008 1 19 05
language	English
source	Enthalten in BMC molecular biology 9(2008), 1 vom: 19. Mai volume:9 year:2008 number:1 day:19 month:05
sourceStr	Enthalten in BMC molecular biology 9(2008), 1 vom: 19. Mai volume:9 year:2008 number:1 day:19 month:05
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Reference Gene Candidate Reference Gene Stable Reference Gene Hypoxanthine Phosphoribosyltransferase Good Reference Gene
isfreeaccess_bool	true
container_title	BMC molecular biology
authorswithroles_txt_mv	Cappelli, Katia @@aut@@ Felicetti, Michela @@aut@@ Capomaccio, Stefano @@aut@@ Spinsanti, Giacomo @@aut@@ Silvestrelli, Maurizio @@aut@@ Supplizi, Andrea Verini @@aut@@
publishDateDaySort_date	2008-05-19T00:00:00Z
hierarchy_top_id	326645004
id	SPR027216411
language_de	englisch
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author	Cappelli, Katia
spellingShingle	Cappelli, Katia misc Reference Gene misc Candidate Reference Gene misc Stable Reference Gene misc Hypoxanthine Phosphoribosyltransferase misc Good Reference Gene Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization
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topic_title	Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization Reference Gene (dpeaa)DE-He213 Candidate Reference Gene (dpeaa)DE-He213 Stable Reference Gene (dpeaa)DE-He213 Hypoxanthine Phosphoribosyltransferase (dpeaa)DE-He213 Good Reference Gene (dpeaa)DE-He213
topic	misc Reference Gene misc Candidate Reference Gene misc Stable Reference Gene misc Hypoxanthine Phosphoribosyltransferase misc Good Reference Gene
topic_unstemmed	misc Reference Gene misc Candidate Reference Gene misc Stable Reference Gene misc Hypoxanthine Phosphoribosyltransferase misc Good Reference Gene
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title	Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization
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title_full	Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization
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author_browse	Cappelli, Katia Felicetti, Michela Capomaccio, Stefano Spinsanti, Giacomo Silvestrelli, Maurizio Supplizi, Andrea Verini
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author-letter	Cappelli, Katia
doi_str_mv	10.1186/1471-2199-9-49
title_sort	exercise induced stress in horses: selection of the most stable reference genes for quantitative rt-pcr normalization
title_auth	Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization
abstract	Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. © Cappelli et al; licensee BioMed Central Ltd. 2008
abstractGer	Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. © Cappelli et al; licensee BioMed Central Ltd. 2008
abstract_unstemmed	Background Adequate stress response is a critical factor during athlete horses' training and is central to our capacity to obtain better performances while safeguarding animal welfare. In order to investigate the molecular mechanisms underlying this process, several studies have been conducted that take advantage of microarray and quantitative real-time PCR (qRT-PCR) technologies to analyse the expression of candidate genes involved in the cellular stress response. Appropriate application of qRT-PCR, however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. Results The expression of nine potential reference genes was evaluated in lymphocytes of ten endurance horses during strenuous exercise. These genes were tested by qRT-PCR and ranked according to the stability of their expression using three different methods (implemented in geNorm, NormFinder and BestKeeper). Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyltransferase (HPRT) always ranked as the two most stably expressed genes. On the other hand, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transferrin receptor (TFRC) and ribosomal protein L32 (RPL32) were constantly classified as the less reliable controls. Conclusion This study underlines the importance of a careful selection of reference genes for qRT-PCR studies of exercise induced stress in horses. Our results, based on different algorithms and analytical procedures, clearly indicate SDHA and HPRT as the most stable reference genes of our pool. © Cappelli et al; licensee BioMed Central Ltd. 2008
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title_short	Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization
url	https://dx.doi.org/10.1186/1471-2199-9-49
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author2	Felicetti, Michela Capomaccio, Stefano Spinsanti, Giacomo Silvestrelli, Maurizio Supplizi, Andrea Verini
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Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization

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