Role of FKBP51 in the modulation of the expression of the corticosteroid receptors in bovine thymus following glucocorticoid administration

Cucuzza, L. Starvaggi; Biolatti, B.; Scaglione, Frine Eleonora; Cannizzo, Francesca Tiziana

doi:10.1016/j.domaniend.2017.08.001

Cited by 6 publications

(5 citation statements)

References 35 publications

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“…FKBP5 gene expression profile detected in this study by ddPCR in thoracic thymus was comparable to previous results obtained using qPCR technology [14], as well as specificity and sensitivity parameters described in analogue qPCR screening tests [23,42].…”

Section: Plos Onesupporting

confidence: 89%

“…This study was conducted on thoracic thymus from bovines experimentally treated with GC and collected at a slaughterhouse. Three trials were conducted as previously described [14,[21][22][23].…”

Section: Experimental Design and Sample Collectionmentioning

confidence: 99%

“…Moreover, samples of thoracic thymus obtained from each animal at slaughterhouse were immediately frozen in liquid nitrogen and then stored at -80˚C for molecular analyses as previously described [14,26].…”

Section: Experimental Design and Sample Collectionmentioning

confidence: 99%

“…For this reason, a transcriptomic approach was previously proposed [13] and clear transcriptomic changes were observed in peripheral blood mononuclear cells of young bulls treated with prednisolone. Subsequently, aberrant FKBP prolyl isomerase 5 (FKBP5) gene expression was reported in the thymus and skeletal muscle of veal calves and young bulls following treatment with prednisolone, allowing the development of an indirect screening test based on FKBP5 expression regulation by GCs [14,15].…”

Section: Introductionmentioning

confidence: 99%

“…In previous studies [14,15], FKBP5 expression was evaluated using quantitative real-time PCR (qPCR) which currently represents the method of choice for this analysis. Transcript measurement can be assessed through an absolute or relative quantification, but they require a standard curve or a calibrator, respectively [16].…”

Section: Introductionmentioning

confidence: 99%

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Development of a droplet digital PCR assay to detect illicit glucocorticoid administration in bovine

et al. 2022

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Glucocorticoids are often used illegally in food-producing animals for the growth promotion of livestock animals. In accordance to official chemical methods for glucocorticoid detection, an animal is declared as non-compliant when a residue is identified in the sample. Neverthless, growth promoting molecules can often escape identification due to their rapid elimination or due to the use of non-detectable new generation drugs. Therefore, an indirect screening method able to detect the biological effect of long-term administration of low doses of dexamethasone and prednisolone on livestock has been developed to support official methods. As already described, FKBP5 (FKBP prolyl isomerase 5) expression in bovine thymus is regulated by glucocorticoids, and this specific regulation can be exploited in an indirect screening assay. In the present study, male veal calves and young bulls were considered in three different trials in which estradiol, dexamethasone, and prednisolone were administered alone or in combination with Revalor-200 subcutaneous pellets. Thoracic thymus was sampled from all animals and molecular analysis was performed. A duplex droplet digital PCR assay with EvaGreen® was employed to detect the target gene expression using absolute quantification. The developed droplet digital PCR assay was precise, showing intra- and inter-assay mean coefficient of variation values of about 6.16% and 3.17%, respectively. It was also highly specific (100%) with Youden’s index of 76.92% and 53.57% applied to veal calves and young bulls, respectively. The lowest detection limit in which the target gene expression level was kept constant, was 0.05 ng/μl of cDNA with 1 copies/μL and 0.5 copies/μL for target and reference gene, respectively. This study establishes the basis for using a digital PCR-based assay as an efficient test to identify animals illegally treated with glucocorticoids.

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Section: Plos Onesupporting

confidence: 89%