10.1021/acs.jproteome.5b00520.s001
Diba Ahmadi Rastegar
Diba Ahmadi
Rastegar
Mehdi Sharifi Tabar
Mehdi Sharifi
Tabar
Mehdi Alikhani
Mehdi
Alikhani
Pouria Parsamatin
Pouria
Parsamatin
Fazel Sahraneshin
Samani
Fazel
Sahraneshin
Samani
Marjan Sabbaghian
Marjan
Sabbaghian
Mohammad
Ali Sadighi Gilani
Mohammad
Ali Sadighi
Gilani
Ali Mohammad Ahadi
Ali Mohammad
Ahadi
Anahita Mohseni
Meybodi
Anahita
Mohseni
Meybodi
Abbas Piryaei
Abbas
Piryaei
Naser Ansari-Pour
Naser
Ansari-Pour
Hamid Gourabi
Hamid
Gourabi
Hossein Baharvand
Hossein
Baharvand
Ghasem Hosseini Salekdeh
Ghasem Hosseini
Salekdeh
Isoform-Level Gene Expression Profiles of Human Y
Chromosome Azoospermia Factor Genes and Their X Chromosome Paralogs
in the Testicular Tissue of Non-Obstructive Azoospermia Patients
American Chemical Society
2015
WB
5D
5C
41 alternative transcripts
PRY
CDY
USP 9Y DDX 3Y XKRY
HSFY
SCOS
IHC
PCR
Human Y Chromosome Azoospermia Factor Genes
NOA
sperm retrieval
28 premiotic maturation arrest
expression
X Chromosome Paralogs
AZFc region genes
1A
RBMX
KDM
X chromosome homologue transcripts
CYORF
12 MA patients
Y chromosome genes
TESE
EIF
BPY
isoform level signature
testicular sperm extraction
RPS
15B
4Y
spermatogenesi
protein level
X chromosome counterparts
DAZL 1 transcripts
1AY
testis cell types
RBMY
azoospermic men
15A
2015-09-04 00:00:00
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Isoform_Level_Gene_Expression_Profiles_of_Human_Y_Chromosome_Azoospermia_Factor_Genes_and_Their_X_Chromosome_Paralogs_in_the_Testicular_Tissue_of_Non_Obstructive_Azoospermia_Patients/2135281
The
human Y chromosome has an inevitable role in male fertility
because it contains many genes critical for spermatogenesis and the
development of the male gonads. Any genetic variation or epigenetic
modification affecting the expression pattern of Y chromosome genes
may thus lead to male infertility. In this study, we performed isoform-level
gene expression profiling of Y chromosome genes within the azoospermia
factor (AZF) regions, their X chromosome counterparts, and few autosomal
paralogues in testicular biopsies of 12 men with preserved spermatogenesis
and 68 men with nonobstructive azoospermia (NOA) (40 Sertoli-cell-only
syndrome (SCOS) and 28 premiotic maturation arrest (MA)). This was
undertaken using quantitative real-time PCR (qPCR) at the transcript
level and Western blotting (WB) and immunohistochemistry (IHC) at
the protein level. We profiled the expression of 41 alternative transcripts
encoded by 14 AZFa, AZFb, and AZFc region genes <i>(USP9Y</i>, <i>DDX3Y</i>, <i>XKRY</i>, <i>HSFY1</i>, <i>CYORF15A, CYORF15B, KDM5D, EIF1AY</i>, <i>RPS4Y2</i>, <i>RBMY1A1</i>, <i>PRY, BPY2, DAZ1</i>, and <i>CDY1</i>) as well as their X chromosome homologue transcripts
and a few autosomal homologues. Of the 41 transcripts, 18 were significantly
down-regulated in men with NOA when compared with those of men with
complete spermatogenesis. In contrast, the expression of five transcripts
increased significantly in NOA patients. Furthermore, to confirm the
qPCR results at the protein level, we performed immunoblotting and
IHC experiments (based on 24 commercial and homemade antibodies) that
detected 10 AZF-encoded proteins. In addition, their localization
in testis cell types and organelles was determined. Interestingly,
the two missing proteins, XKRY and CYORF15A, were detected for the
first time. Finally, we focused on the expression patterns of the
significantly altered genes in 12 MA patients with successful sperm
retrieval compared to those of 12 MA patients with failed sperm retrieval to predict the success of sperm retrieval in azoospermic men. We showed that <i>HSFY1-1</i>, <i>HSFY1-3</i>, <i>BPY2-1</i>, <i>KDM5C2</i>, <i>RBMX2, </i>and <i>DAZL1</i> transcripts could be used as potential molecular markers to predict the presence of spermatozoa in MA patients. In this study, we have identified isoform level signature that can be used to discriminate effectively between MA, SCOS, and normal testicular tissues and suggests the possibility of diagnosing the presence of mature sperm cell in azoospermic men to prevent additional testicular sperm extraction (TESE) surgery.