Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia.

Abstract

PURPOSE:

Non-obstructive azoospermia (NOA) is one the many causes of male infertility (10 %) resulting from testicular failure. Multiple testicular biopsies fail to find mature sperm in at least 50 % of cases Therefore; hunting for sensitive and specific biomarkers of spermatogenesis that could better determine the fertility status in NOA can lead to improved management of male infertility. Therefore, we evaluated sperm production through analyses of germ cell-specific transcripts (DAZ, TSPY1, SPTRX3 and SPTRX1) in semen and testicular biopsies of men with azoospermia.

METHODS:

We collected semen (N=83) and testis biopsies (N=31) from men with non-obstructive azoospermia. We later extracted RNA and synthesized cDNA using washed semen precipitate and testicular tissues. We also performed semi-nested PCR with designed specific primers. Using H&E method, an expert pathologist performed the histopathological evaluation. Having categorized the patients into three groups based on histopathological results, we calculated the agreement between molecular results of semen and tissues with histopathological findings for each patient using Kappa statistical test.

RESULTS:

Molecular findings of precipitated semen and testicular tissues were in disagreement with histopathological results in most cases. Molecular analysis of testis biopsies showed significant difference (Kappa coefficient=0.009, P value=0.894) with histopathological results; TSPY1, DAZ, SPTRX3 and SPTRX1 were respectively detected in 94 %, 94 %, 17.6 % and 52.9 % of men diagnosed with germ cell aplasia.

CONCLUSIONS:

Molecular analysis of semen does not provide sufficient sensitivity and specificity to be used as a screening test at the present time, but it is a useful adjunct to histopathological methods in men with NOA. Spermatid/sperm specific transcripts indicated the possibility to find mature sperm following repeated multiple testicular sperm extraction (TESE) or microdisection TESE (mTESE).

• Authors: Maryam Eghbali, Mohammad Reza Sadeghi, Niknam Lakpour, Hale Edalatkhah, Hojjat Zeraati, Haleh Soltanghoraee, Mohammad Mehdi Akhondi, S Behnam Hashemi, Mohammad Hossein Modarressi
• Journal: Journal of assisted reproduction and genetics (J Assist Reprod Genet) Vol. 31 Issue 6 Pg. 707-15 (Jun 2014) ISSN: 1573-7330 [Electronic] Netherlands
• PMID: 24728569 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Cell Cycle Proteins
• DAZ1 protein, human
• Deleted in Azoospermia 1 Protein
• Membrane Proteins
• RNA-Binding Proteins
• TSPY1 protein, human
• TXNDC2 protein, human
• TXNDC8 protein, human
• Thioredoxins

Topics

• Adult
• Azoospermia (genetics, pathology)
• Biopsy
• Cell Cycle Proteins (biosynthesis)
• Deleted in Azoospermia 1 Protein
• Gene Expression Regulation, Developmental
• Humans
• Infertility, Male (genetics, pathology)
• Male
• Membrane Proteins (biosynthesis)
• RNA-Binding Proteins (biosynthesis)
• Semen (cytology)
• Spermatogenesis (genetics)
• Spermatozoa (pathology)
• Testis (metabolism, pathology)
• Thioredoxins (biosynthesis)