the relationship between the AZF microdeletion regions and the azoospermia phenotype.

1. INSIGHT INTO AZF REGION OF Y
CHROMOSOME AND MALE
INFERTILITY
Presented By: Shalaka Chitale
Presented On: 3rd December, 2016
Trainee
GeneXplore Diagnostic and Research Centre Pvt.
Ltd.
1

2. Y - Chromosome
2
source: humupd.oxfordjournals.org
PAR: pseudoautosomal
region
SRY: Sex Determining
Region
MSY: Male Specific Region
AZF: Azoospermia Factor
Yq
11.1
to
11.223
MSY

3. Hormones
3
 FSH – Follicle Stimulating Hormone
 LH/ISCH – Interstitial Cell Stimulating Hormone
 ABP – Androgen Binding Protein
 GnRH – Gonadtropin Releasing Hormone
 GnIH – Gonadotropin Inhibiting Hormone
 Inhibin
 Testosterone

4. Mechanism of LH(Luteinizing Hormone) and
FSH (Follicle Stimulating Hormone) for
Spermatogenesis
source:
http://www.austincc.edu/
4

5. Clinical Characteristics and Treatment of
Azoospermia and Severe Oligospermia Patients
With Y-Chromosome Microdeletions
5
FENGBIN ZHANG, LEJUN LI, LIQUAN WANG, LIMING YANG, ZHONGYAN
LIANG, JINGPING LI, FAN JIN, AND YONGHONG TIAN
Department of Reproductive Endocrinology, Women’s Hospital, School of Medicine,
Reproductive Medicine Center, Zhejiang University, Hangzhou, Zhejiang, China
Journal : Molecular Reproduction & Development, 2013 (2.67)
D.O.I : 10.1002/mrd.22226

6. Overview
6
 Introduction
 Methods
 Results
 Discussion

7. Introduction
 A comparative analysis of males to evaluate the
relationship between the AZF microdeletion regions and
the azoospermia phenotype.
 In this study, analysis of the relationships between the
AZF microdeletion region and its clinical manifestations,
testicular pathological features, serum levels of
reproductive hormones, and the subsequent assisted
reproductive treatment.
 The AZF loci have three non-overlapping regions: AZFa,
AZFb, and AZFc. Deletions within each region are
associated with clinically distinct phenotypes.
7

8. 8
 The AZFa deletion is rare (0.28% among non-
obstructive azoospermia), and is associated with
azoospermia and an absence of sperm cells in testicular
tissue.
 Deletion of the entire AZFb interval usually results in
maturation arrest during meiosis.
 Complete deletion of the AZFc region, the most common
genotype, resulted in a variable phenotype ranging from
hypospermatogenesis, to spermatogenic arrest, to
Sertoli cell-only syndrome (SCOS).

9. Methods
 120 patients with azoospermia and severe oligospermia
(AZF microdeletion) and 50 control patients
 Clinical examinations like Hormone analysis, semen
analysis, detection of AZF microdeletions, testicular
biopsy and statistical analysis.
 Healthy fertile men and women were used for positive
and negative controls, respectively.
9

10. 10
 Genomic DNA samples were extracted from peripheral blood
lymphocytes using a DNA-isolation kit (TaKaRa Co., Dalian,
China)
 Sequence-tagged site (STS) markers in the azoospermic AZF
region detected by PCR
 STS was absent after three amplification failures against
successful internal control amplification.
AZFa sY84, sY86
AZFb sY127, sY134
AZFc sY254, sY255
Preheating
(95°C for 5 min)
95°C for 1
min
35x
56°C for 30
sec
35x
72°C for 1 min
35x
Extension
(72°C for 7
min)

11. Results
11

12. Johnsen
Score
12

13. 13
Pathological Characteristics and Johnsen Score of
Testicular Specimen in 34 Patients Undergoing Biopsy

14. A: AZFa+b+c: germ-cell aplasia (100x
magnification); B: AZFa+b+c: Sertoli cell-only
syndrome (200x magnification); C: AZFb:
spermatid arrest (400x magnification); D: AZFc:
hypospermatogenesis (400x magnification);
E: AZFc: primary spermatocyte arrest (400x
magnification);
F: AZFc: secondary spermatocyte arrest (400x
magnification). Scale bars represent lengths
shown.
14
Histological section of testis biopsy with some
AZF deletion patients.

15. Reproductive Hormone Levels in Patients With
AZF Microdeletions
15

16. Discussion
16
 120 patients with Y-chromosome microdeletion
revealed that 64% of the microdeletions were located
in the AZFc locus.
 The AZFb+c joint microdeletion in 20% of the patients
in this study, and the AZFb deletion in 13% of these
patients; other types of microdeletion were rare.
 Here, only 2 (1.7%) patients out of 120 carried AZFa
deletion, AZFa deletion results in spermatogenic failure
and altered reproductive hormone production and so
testicular sperm retrieval technologies such as TESA or
m-TESE are not advised. Instead, AID is
recommended.

17. 17
 Deletions in the AZFb region manifest
spermatogenic arrest, mainly at the
spermatocyte or sperm-cell development
stages, and these patients present clinically with
azoospermia or severe oligospermia.

18. Clinical Treatment and Outcome of the Azoospermia
Cases With an AZFb or AZFc Microdeletion
18

19. 19
 Reproductive hormone testing is of clinical significance for the
diagnosis of oligospermia and azoospermia patients and for
planning therapeutic treatment strategies.
 The average level of serum LH and FSH levels in patients
with a Y-chromosome microdeletion were significantly higher
than fertile-subject levels, but 73% (11/15) of patients with
AZFb microdeletion and39% (30/77) of patients with AZFc
microdeletion had normal serum FSH levels. So, reproductive
hormone level cannot be regarded as the sole indicator of
AZF microdeletion.
 AZFb- or AZFcmicrodeletion azoospermia patients with
relatively normal testicular volumes, residual sperm should be
retrieved byTESE or m-TESE, followed by ICSI where male
embryos should be removed by PGD to avoid genetic risks.

20. 20