Potential biomarkers of nonobstructive azoospermia identified in microarray gene expression analysis
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Published on: November 2, 2013
Agnieszka Malcher1, Natalia Rozwadowska, Tomasz Stokowy
1Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.
Objective:
To identify potential biomarkers of azoospermia to determine a particular stage of spermatogenetic differentiation.
Design:
GeneChip Human Gene 1.0 ST microarray with validation at mRNA and protein levels.
Setting:
Basic research laboratory.
Patient(S):
Men with various types of nonobstructive azoospermia (n = 18) and with normal spermatogenesis (n = 4).
Intervention(S):
Obtaining 31 testicular biopsy samples.
Main Outcome Measure(S):
Gene expression analysis using the Affymetrix Human Gene 1.0 ST microarrays on 14 selected genes according to the highest fold change, verified with quantitative polymerase chain reaction and on independent set of microarray samples. Western blot and immunohistochemistry were additionally performed.
Result(S):
The comparative analysis of gene expression profiles in the infertile and control groups resulted in the selection of 4,946 differentially expressed genes. AKAP4, UBQLN3, CAPN11, GGN, SPACA4, SPATA3, and FAM71F1 were the most significantly down-regulated genes in infertile patients. Global analysis also led to identification of up-regulated genes-WBSCR28, ADCY10, TMEM225, SPATS1, FSCN3, GTSF1L, and GSG1-in men with late maturation arrest. Moreover, the results from quantitative polymerase chain reaction and Western blot largely confirmed the microarray data.
Conclusion(S):
The set of selected genes can be used to create a molecular diagnostic tool to determine the degree of spermatogenic impairment for men with idiopathic nonobstructive azoospermia.
