Quantification of cyclin A1 and glyceraldehyde-3-phosphate dehydrogenase expression in testicular biopsies of infertile patients by fluorescence real-time RT-PCR.
The objective of this study was to establish a classification of meiotic disorders by detection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and cyclin A1 mRNA in patients presenting with non-obstructive azoospermia. GAPDH and cyclin A1 expression levels were detected in 37 histologically classified testicular tissue specimens by one-line RT-PCR. Levels of cyclin A1 expression (NCyclinA1) were high in tissue specimens with full spermatogenesis [3.519 +/- 3.141 (mean +/- SD)] but only minimal in those without haploid germ cells. A lack of cyclin A1 expression was seen in most sertoli-cell-only syndrome (SCOS) specimens. The criteria for an ideal internal standard were only partially met by GAPDH, as we detected decreased expression in tissue samples with maturation arrest [(268.2 +/- 106.2 relative gene expression (mean +/- SD)] and SCOS (201.7 +/- 95.2) compared to those with normal histological findings (325.1 +/- 129.3). It was concluded that the level of cyclin A1 mRNA expression predicts meiotic disorders during spermatogenesis. GAPDH is not an ideal internal standard for specific gene expression in testicular tissue specimens.
Department of Urology, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Berlin, Germany. schrader@medizin.fu-berlin.de
Mark
M
Schrader
Stuart
S
Ravnik
Carsten
C
Müller-Tidow
Markus
M
Müller
Bernd
B
Straub
Sven
S
Diedrichs
Hubert
H
Serve
Kurt
K
Miller
D000328:Adult; D001706:Biopsy; D019925:Cyclin A / Q000096:biosynthesis* / Q000235:genetics; D056750:Cyclin A1; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis*; D006801:Humans; D007248:Infertility, Male / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000473:pathology; D013091:Spermatogenesis; D013737:Testis / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology
C503745:CCNA1 protein, human; D019925:Cyclin A; D056750:Cyclin A1; D012333:RNA, Messenger; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases