Quantification of telomerase activity, porphobilinogen deaminase and human telomerase reverse transcriptase mRNA in testicular tissue – new parameters for a molecular diagnostic classification of spermatogenesis disorders

Abstract

The aim of the study was to evaluate the quantitative detection of human telomerase reverse transcriptase (hTERT) mRNA and telomerase activity as new molecular diagnostic parameters for a subclassification of spermatogenesis disorders. Telomerase activity was detected by a quantitative telomerase PCR ELISA, and hTERT mRNA expression was quantified by fluorescence real-time RT-PCR in a LightCycler in cryopreserved testicular tissue specimens. This was paralleled by a histological workup. The discriminant analysis showed that detection of normalized hTERT expression was able to correctly classify 89.0% of the investigated tissue specimens into the subgroups of full spermatogenesis, maturation arrest or Sertoli-cell-only syndrome. In contrast, discriminant analysis revealed an only 58% accuracy of telomerase activity for the investigated tissue specimens. This study shows that the quantification of hTERT expression in testicular tissue by real-time fluorescence RT-PCR is well suited for correctly classifying spermatogenesis disorders and proved to be markedly superior to the determination of telomerase activity.

The differentiation status of primary gonadal germ cell tumors correlates inversely with telomerase activity and the expression level of the gene encoding the catalytic subunit of telomerase

“…Expression of hTERT mRNA ( N hTERT = 132.0 (± 48.0)) proved to be highest in tissue samples with normal gametogenesis but was significantly reduced in those with maturation arrest at the spermatocyte I° and II° level ( N hTERT = 53.0 (± 17.3)) and at the level of spermatogonia ( N hTERT = 23.2 (± 7.8)), as described previously [34]. In patients with germ cell aplasia, hTERT mRNA expression was absent in 6 of 8 cases and only minimal in two with marked granulocytic infiltration ( N hTERT = 7.2 and 4.1), indicating that, hTERT expression is germ-cell-specific in benign testicular tissue.…”

Quantification of survivin mRNA in testes of infertile patients and in testicular germ cell tumours: high levels of expression associated with normal spermatogenesis

“…The median (range) PBGD expression was 10 486 copies (3495–87 530) in tissue specimens with SCOS, 35 790 copies (7609–81 770) in those pre‐meiotic maturation arrest, 56 120 copies (3821–107 300) in those with post‐meiotic maturation arrest, 30 600 copies (5901–201 800) in those with normal spermatogenesis, and 21 780 copies (163–412 000) in tumour samples. Comparisons of individual groups disclosed significant differences between SCOS and normal spermatogenesis ( p = .031), so that the criteria for an ideal internal standard were only partially met by PBGD as shown previously (Schrader et al. , 2002a).…”

DMRTC2, PAX7, BRACHYURY/T and TERT Are Implicated in Male Germ Cell Development Following Curative Hormone Treatment for Cryptorchidism-Induced Infertility

“…Atrophic testes had lower TERT expression levels relative to normal testes, leading to the suggestion that telomerase plays a role in maintaining germ cell proliferation [ 50 ]. Furthermore, the TERT mRNA expression level was shown to be effective in both classifying spermatogenesis disorders in patients, and in predicting successful sperm recovery in azoospermia patients [ 51 , 52 ]. Androgens were reported to enhance TERT expression in human primary hematopoietic cells [ 53 ].…”