Expression stability of putative reference genes in equine endometrial, testicular, and conceptus tissues.
Abstract: Quantitative RT-PCR data are commonly normalized using a reference gene. A reference gene is a transcript which expression does not differ in the tissue of interest independent of the experimental condition. The objective of this study was to evaluate the stability of mRNA expression levels of putative reference genes in three different types of equine tissue, endometrial, testicular, and conceptus tissue. Results: The expression stability of four (uterine tissue) and six (testicular and conceptus tissue) was assessed using descriptive data analysis and the software programs Normfinder and geNorm. In uterine samples, 18S showed the largest degree of variation in expression while GAPDH, B2M, and ACTB were stably expressed. B2M and GAPDH were identified as the most stably expressed genes in testicular samples, while 18S showed some extent of regulation between samples. Conceptus tissue overall was characterized by very low variability of the transcripts analyzed with GAPDH, YWHZ, and 18S being the most stably expressed genes. Conclusions: In equine endometrium, GAPDH, B2M, and ACTB transcript levels are equally stable, while 18S is less stably expressed. In testes and associated structures, B2M and GAPDH are the transcripts showing the least amount of variation, while in conceptus tissue GAPDH, YWHZ, and 18S were identified as the most suitable reference genes. Overall, transcripts analyzed in conceptus tissue were characterized by less variation than transcripts analyzed in uterine and testicular tissue.
Publication Date: 2011-04-12 PubMed ID: 21486450PubMed Central: PMC3083352DOI: 10.1186/1756-0500-4-120Google Scholar: Lookup
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- Journal Article
Summary
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The research is about finding the stability of mRNA expression levels of potential reference genes in three different types of horse tissue, specifically endometrial, testicular, and conceptus tissue. Their findings suggest that in equine endometrium, GAPDH, B2M, and ACTB transcript levels are equally stable, while 18S is less stably expressed. In testes and associated structures, B2M and GAPDH are the least variable, while in conceptus tissue GAPDH, YWHZ, and 18S were identified as the most suitable reference genes.
Objective of the Study
- The main objective of this study was to assess the stability of mRNA expression levels of potential reference genes in different equine tissues – endometrial, testicular, and conceptus. As reference genes are used to normalize quantitative RT-PCR data, their expression stability is critical for accurate results.
Methods of the Study
- The researchers evaluated the expression stability of four putative reference genes for uterine tissue and six for testicular and conceptus tissues.
- Descriptive data analysis was used for this evaluation along with software programs such as Normfinder and geNorm.
Results
- In uterine samples, the gene 18S showed the highest degree of variation while GAPDH, B2M, and ACTB were more stably expressed.
- For testicular samples, B2M and GAPDH showed the most stable expression.
- In conceptus tissue, the transcripts that were analyzed showed very low variability with GAPDH, YWHZ, and 18S being the genes with the most stable expression.
Conclusion
- The study concludes that the genes GAPDH, B2M, and ACTB have equally stable transcript levels in equine endometrium and the gene 18S is less stably expressed.
- In the case of testicular tissue and its associated parts, the genes B2M and GAPDH were found to be the most stable while 18S showed some variation between samples.
- In conceptus tissue, the most suitable reference genes were identified as GAPDH, YWHZ, and 18S.
- The transcripts analyzed in conceptus tissue showed less variation as compared to those in uterine and testicular tissues.
Cite This Article
APA
Klein C, Rutllant J, Troedsson MH.
(2011).
Expression stability of putative reference genes in equine endometrial, testicular, and conceptus tissues.
BMC Res Notes, 4, 120.
https://doi.org/10.1186/1756-0500-4-120 Publication
Researcher Affiliations
- University of Kentucky, Department of Veterinary Science, 108 Gluck Equine Research Center, Lexington, KY, 40546, USA. claudia.klein@uky.edu.
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