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- Title
Selective measurement of α smooth muscle actin: why β-actin can not be used as a housekeeping gene when tissue fibrosis occurs.
- Authors
Veres‑Székely, Apor; Pap, Domonkos; Sziksz, Erna; Jávorszky, Eszter; Rokonay, Réka; Lippai, Rita; Tory, Kálmán; Fekete, Andrea; Tulassay, Tivadar; Szabó, Attila J.; Vannay, Ádám
- Abstract
Background: Prevalence of fibroproliferative diseases, including chronic kidney disease is rapidly increasing and has become a major public health problem worldwide. Fibroproliferative diseases are characterized by increased expression of α smooth muscle actin (α-SMA) that belongs to the family of the six conserved actin isoforms showing high degree homology. The aim of the present study was to develop real-time PCRs that clearly discriminate α-SMA and β-actin from other actin isoforms. Results: Real-time PCRs using self-designed mouse, human and rat specific α-SMA or β-actin primer pairs resulted in the specific amplification of the artificial DNA templates corresponding to mouse, human or rat α-SMA or β-actin, however β-actin showed cross-reaction with the housekeeping γ-cyto-actin. We have shown that the use of improperly designed literary primer pairs significantly affects the results of PCRs measuring mRNA expression of α-SMA or β-actin in the kidney of mice underwent UUO. Conclusion: We developed a set of carefully designed primer pairs and PCR conditions to selectively determine the expression of mouse, human or rat α-SMA and β-actin isoforms. We demonstrated the importance of primer specificity in experiments where the results are normalized to the expression of β-actin especially when fibrosis and thus increased expression of α-SMA is occur.
- Subjects
SMOOTH muscle diseases; KIDNEY disease diagnosis; HOUSEKEEPERS; DISEASE prevalence; HOMOLOGY theory
- Publication
BMC Molecular Biology, 2017, Vol 18, p1
- ISSN
1471-2199
- Publication type
Article
- DOI
10.1186/s12867-017-0089-9