Expression pattern of germ cell-specific genes in the testis of patients with nonobstructive azoospermia: usefulness as a molecular marker to predict the presence of testicular sperm.
OBJECTIVE
To analyze the expression pattern of testis-specific genes of patients with various spermatogenic defects and their usefulness as a molecular marker to predict the presence of testicular spermatozoa in patients with nonobstructive azoospermia undergoing IVF.


METHODS
Prospective, controlled study.


METHODS
Hospital-based infertility research laboratory.


METHODS
Fifty-eight men with azoospermia or severe oligozoospermia.


METHODS
Testicular biopsy was done in the patients with obstructive or nonobstructive azoospermia, including Sertoli cell-only syndrome, maturation arrest, severe hypospermatogenesis, and normal spermatogenesis.


METHODS
Reverse transcriptase polymerase chain reaction was performed using 1 microgram of total RNA extracted from testicular tissues. Three pairs of primers were used for amplification of male germ cell-specific genes (DAZ, transcribed in male germ cells; PGK2, in late spermatocytes and spermatids; protamine-2, in spermatids) as molecular markers. Testicular sperm was obtained by multiple testicular sperm extraction.


RESULTS
The DAZ, PGK2, and protamine-2 genes were expressed in 38, 30, and 21 of the 43 patients with nonobstructive azoospermia, respectively. Testicular spermatozoa were successfully extracted in 4 of 43 patients with nonobstructive azoospermia with the use of multiple testicular sperm extraction. Detection of protamine-2 transcripts predicted the presence or absence of spermatozoa in the testicular tissue in 39 of 43 patients (91%).


CONCLUSIONS
Expression of the protamine-2 gene may be a useful molecular marker to predict the presence of testicular sperm in patients with nonobstructive azoospermia.
Song
G J
GJ
Samsung Cheil Hospital and Women's Health Care Center, Sungkyunkwan University School of Medicine, Seoul, Korea, South Korea. phdsong@samsung.co.kr
Lee
H
H
Park
Y
Y
Lee
H J
HJ
Lee
Y S
YS
Seo
J T
JT
Kang
I S
IS
D000328:Adult; D000075364:Deleted in Azoospermia 1 Protein; D015870:Gene Expression; D005819:Genetic Markers; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009845:Oligospermia / Q000235:genetics*; D010735:Phosphoglycerate Kinase / Q000235:genetics; D011446:Prospective Studies; D011479:Protamines / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D013091:Spermatogenesis; D013094:Spermatozoa / Q000502:physiology*; D013737:Testis / Q000503:physiopathology*
C095668:DAZ1 protein, human; D000075364:Deleted in Azoospermia 1 Protein; D005819:Genetic Markers; D007527:Isoenzymes; D011479:Protamines; D016601:RNA-Binding Proteins; C068621:protamine 2; D010735:Phosphoglycerate Kinase; C104317:phosphoglycerate kinase, testis specific