A biopsy sample reduction approach to identify significant alterations of the testicular transcriptome in the presence of Y-chromosomal microdeletions that are independent of germ cell composition

TY - JOUR

T1 - A biopsy sample reduction approach to identify significant alterations of the testicular transcriptome in the presence of Y-chromosomal microdeletions that are independent of germ cell composition

AU - Cappallo-Obermann, Heike

AU - von Kopylow, Kathrein

AU - Schulze, Wolfgang

AU - Spiess, Andrej-Nikolai

PY - 2010/10/1

Y1 - 2010/10/1

N2 - Y-chromosomal microdeletions (YCMD) are the major genetic cause of male infertility. To date, it is not known which global changes are induced by the presence of AZFc or AZFb+c deletions in the human testicular transcriptome. We investigated this question by microarray analysis in which we had to eliminate the 'germ cell effect', i.e., the dominating effect of germ cell transcripts due to the quantitative difference in germ cell composition in samples with/without YCMD. This problem was tackled by selecting 26 samples from an initial cohort of 34 samples by their homogeneity in respect to cellular composition as obtained from gene expression clustering. This way, the 'germ cell effect' was minimized, and a distinct 'deletion effect' became more apparent. Several hundred genes are influenced by YCMD as shown on the three different phenotypes hypospermatogenesis, meiotic arrest, and Sertoli-cell only syndrome. We validated on an independent cohort of samples five genes by quantitative real-time PCR that are expressed in germ cells or the somatic compartment and which are exclusively altered by the presence of YCMD. We conclude that the deletion of Y-chromosomal genes has a significant effect on spermatogenesis by modulating the transcriptional network of the germ cell and somatic compartment.

AB - Y-chromosomal microdeletions (YCMD) are the major genetic cause of male infertility. To date, it is not known which global changes are induced by the presence of AZFc or AZFb+c deletions in the human testicular transcriptome. We investigated this question by microarray analysis in which we had to eliminate the 'germ cell effect', i.e., the dominating effect of germ cell transcripts due to the quantitative difference in germ cell composition in samples with/without YCMD. This problem was tackled by selecting 26 samples from an initial cohort of 34 samples by their homogeneity in respect to cellular composition as obtained from gene expression clustering. This way, the 'germ cell effect' was minimized, and a distinct 'deletion effect' became more apparent. Several hundred genes are influenced by YCMD as shown on the three different phenotypes hypospermatogenesis, meiotic arrest, and Sertoli-cell only syndrome. We validated on an independent cohort of samples five genes by quantitative real-time PCR that are expressed in germ cells or the somatic compartment and which are exclusively altered by the presence of YCMD. We conclude that the deletion of Y-chromosomal genes has a significant effect on spermatogenesis by modulating the transcriptional network of the germ cell and somatic compartment.

KW - Biopsy

KW - Chromosome Deletion

KW - Chromosomes, Human, Y

KW - Cluster Analysis

KW - Cohort Studies

KW - Gene Expression Profiling

KW - Genetic Loci

KW - Germ Cells

KW - Humans

KW - Infertility, Male

KW - Male

KW - Meiosis

KW - Oligonucleotide Array Sequence Analysis

KW - Seminal Plasma Proteins

KW - Sertoli Cell-Only Syndrome

KW - Spermatogenesis

KW - Testis

U2 - 10.1007/s00439-010-0865-9

DO - 10.1007/s00439-010-0865-9

M3 - SCORING: Journal article

C2 - 20668881

VL - 128

SP - 421

EP - 431

JO - HUM MOL GENET

JF - HUM MOL GENET

SN - 0964-6906

IS - 4

ER -