Quantification of the expression level of the gene encoding the catalytic subunit of telomerase in testicular tissue specimens predicts successful sperm recovery.
BACKGROUND
The objective of the present study was to evaluate the quantitative detection of human telomerase reverse transcriptase (hTERT) mRNA as a new molecular diagnostic parameter in the work-up of testicular tissue specimens from patients presenting with non-obstructive azoospermia. M ETHODS: hTERT mRNA expression was quantified in 49 cryopreserved testicular tissue specimens by fluorescence real-time RT-PCR in a LightCycler. This was paralleled by conventional histological work-up in all tissue specimens and additional semithin sectioning preparation in cases with maturation arrest (n = 20) and Sertoli cell-only syndrome (SCOS; n = 12).


RESULTS
The average normalized hTERT expression (N(hTERT)) was 136.1 +/- 41.7 copies (mean +/- standard deviation) in tissue specimens with presence of haploid germs cells, N(hTERT) = 48.2 +/- 21.0 copies in those with maturation arrest and N(hTERT) = 2.7 +/- 2.8 copies in those with SCOS. The discriminant analysis showed that detection of N(hTERT) was able correctly to classify 89.0% of the investigated tissue specimens.


CONCLUSIONS
Our results demonstrate that quantitative detection of hTERT mRNA expression in testicular tissue enables a molecular-diagnostic subclassification of spermatogenesis disorders. Quantitative detection of hTERT in testicular biopsies is thus well suited for predicting successful sperm recovery in patients with azoospermia and is a useful molecular diagnostic parameter for supplementing the histopathological evaluation.
Schrader
M
M
Department of Urology, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Hindenburgdamm 30, 12200 Berlin, Germany. schrader@medizin.fu-berlin.de
Müller
M
M
Schulze
W
W
Heicappell
R
R
Krause
H
H
Straub
B
B
Miller
K
K
D000328:Adult; D001706:Biopsy; D015925:Cryopreservation; D016002:Discriminant Analysis; D015870:Gene Expression; D006801:Humans; D007248:Infertility, Male / Q000201:enzymology / Q000473:pathology; D008297:Male; D009845:Oligospermia / Q000201:enzymology / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012708:Sertoli Cells / Q000473:pathology; D013091:Spermatogenesis; D013093:Spermatogonia / Q000473:pathology; D013577:Syndrome; D019098:Telomerase / Q000235:genetics*; D013737:Testis / Q000201:enzymology* / Q000473:pathology; D020858:Tissue and Organ Harvesting
D012333:RNA, Messenger; D019098:Telomerase