Hypospermatogenesis Research Articles

Association of Two Polymorphisms in H2B.W Gene with Azoospermia and Severe Oligozoospermia in An Iranian Population.

During spermatogenesis, the H2B family, member W (H2B.W) gene, en- codes a testis specific histone that is co-localized with telomeric sequences and has the potential role to mediate the sperm-specific chromatin remodeling. Previously H2B.W genetic variants were reported to be involved in susceptibility to spermatogenesis im- pairment. In the present study, two single nucleotide polymorphisms (SNPs) in 5΄UTR and exon 1 of H2B.W gene were examined to investigate possible association of these polymorphisms with male infertility in Iranian population. This case control study was conducted in Royan institute during four-year period (2010-2013). Genetic alteration of two SNPs loci, -9C>T and 368A>G, in H2B.W gene were indicated in 92 infertile men who were divided into two main groups includ- ing azoospermia (n=46) and sever oligozoospermia (n=46), while there was 60 fertile men as control group. Azoosperima was also divided into three sub-groups including sertoli cell only syndrome (SCOS, n=21), complete maturation arrest (CMA, n=17) and hypo spermatogenesis (n=8) according to testicular biopsy. For analysis, polymerase chain reaction-restriction frag- ment length polymorphism (PCR-RFLP) technique was applied. The frequency of allele -9T was significantly higher in CMA group than in patients with SCOS (P<0.05). The haplotype TA (corresponding to simultaneous occur- rence of -9T and 368A) compared with haplotype CA (corresponding to simultaneous occurrence of -9C and 368A) in patients suffering from CMA significantly increased, compared with patients had SCOS (P<0.05). However, statistical studies indicated that in general, the distribution frequencies of -9C>T and 368A>G had no significant difference between the infertile groups and control (P=0.859 and P=0.812, respectively). This investigation showed that SNP -9C>T might be contribute to CMA in azoo- spermic patients and SNP 368A>G had no correlation with male infertility in Iranian population.

Primary Testicular Failure: An Overview

Primary testicular failure (PTF) or hypergonadotropic hypogonadism refers to conditions where testes fail to produce sperm despite adequate hormonal support. Primary testicular failure is major cause of non-obstructive azoospermia and oligospermia. It affects approximately 1% of all men and 10% of those seeking fertility evaluations [1]. Non-obstructive azoospermia i.e. defect in spermatogenesis can be primary (testicular pathology) or secondary (pathology outside testes; mostly hypothalamo-pituitary axis or pretesticular). The pre-testicular (hypogonadotropic hypogonadism) causes of azoospermia may be defined as extragonadal endocrine disorder originating in the pineal, hypothalamus, pituitary, or adrenals, which have an adverse effect on spermatogenesis through aberrant hormonal action. The testicular (hypergonadotropic hypogonadism/ primary testicular failure) causes of azoospermia are primary defects of the testes. Primary testicular failure is classified into four distinct subtypes viz., sertoli cell only syndrome (SCOS), germ cell/maturation arrest (GCA/MA), hypospermatogenesis (HS) and tubular fibrosis (TF) according to histopathology/cytology findings although accurate categorization is only possible by multiple testicular biopsies/fine needle aspiration cytology (FNAC). However, often one may find differences in subtypes between testes viz., SCOS in one side and tubular fibrosis in other side or any other combinations and these cases are usually labeled as mixed group. In addition many patients may present as complete germinal cell aplasia in some tubules whereas complete spermatogenesis in adjacent tubules (focal germinal cell aplasia) or some tubules with sertoli cells only/hyaline sclerosis and other tubules with complete spermatogenesis or any combinations. This categorization should be considered as a description of histopathologic phenotypes of spermatogenic failure, and not as manifestations of disease entities. Sometimes, patients may present as hypospermatogenesis/maturation arrest initially and later as sertoli cell only syndrome/testicular fibrosis over a period of few years hence diagnosis may change with time. Sertoli cell only syndrome or germ cell aplasia applies to a testis, in which germ cells at any stage are absent, but the tubular architecture is not affected by fibrosis, and supporting cells continue to be present. Sertoli cells play a central role in development of a functional testis, and hence in the expression of a male phenotype. Sertoli cell only syndrome is a common finding of non-obstructive azoospermia. It is a histopathologic phenotype of spermatogenic failure and described first by Castillo et al. [2], in complete germ cell aplasia, the tubules are reduced in diameter, contain only sertoli cells and no other cells involved in spermatogenesis are present. The primordial germ cells either do not migrate from the yolk sac into the future gonads or do not survive in the seminiferous tubules. Germ cell aplasia can also be focal with a variable percentage of tubules containing germ cells, but even in these tubules, spermatogenesis is often have limited activity [3] and hence, should be categorized as hypospermatogenesis. Maturation/Germ Cell arrest is the interruption of normal germ cell maturation at a specific cell stage including spermatogonia, spermatocyte or spermatid level. In hypospermatogenesis all the stages of spermatogenesis are present in some or all tubules, but they are reduced in number. Tubular fibrosis presents as thickening of peritubular membrane and hyaline deposition on basement membrane with absence of germ cells and sertoli cells.

The assessment of fertilization and embryonic development using testicular spermatozoa among presumed histopathology in testicular biopsy from non-obstructive azoospermia (NOA)

Fertilization and pregnancies can be obtained with spermatozoa recovered from the seminiferous tubules. Sertoli cell only syndrome (SCOS), maturation arrest (MA), and hypospermatogenesis (HS), and, with or without focal spermatogenesis are the commonest histological patterns of patients with non-obstructive azoospermia (NOA). To our knowledge, no study has specifically examined the results of intracytoplasmic sperm injection (ICSI) in SCOS, MA, and HS patients. The aim of this study is to assess sperm retrieval rates (SRR) by microdissection testicular sperm extraction (micro-TESE), fertilization rate, and embryonic development among patients with presumed SCOS, MA, and HS in testicular biopsy in those couples whom spermatozoa were obtained.

Testicular FNAC in Azoospermia

Aim: To describe the technique and findings of fine needle aspiration cytology (FNAC) of the testes for the assessment of azoospermic male infertility.Methods: Fifty four azoospermic patients were included in this study. A single dose of diclofenac sodium (50mg) Supository was given per rectally half an hour before aspiration. The FNA was done on both testes under surface anesthesia with 23G needle attached to a 10ml disposable syringe. Aspirated smears were stained with H&amp;E.Results: Normal spermatogenesis in both testes was found in 6(11.11%), only in right testis 03(5.55%) and only in left testis 03(5.55%) cases. Hypo spermatogenesis in 07(12.96%) cases . Sertoli cell syndrome in 05(9.26%) and maturation arrest in 30 (55.60%) cases. No significant complications were noted.Conclusion: FNAC of testis is a simple, well tolerated and informative procedure in assesing azoospermia.DOI: http://dx.doi.org/10.3329/cmoshmcj.v13i1.19419

Expression analysis of MND1/GAJ, SPATA22, GAPDHS and ACR genes in testicular biopsies from non-obstructive azoospermia (NOA) patients

BackgroundHigh-throughput studies provide a wide spectrum of genes for use as predictive markers during testicular sperm extraction (TESE) in combination with ICSI. In this work, we used the specimens from testicular biopsies of men with non-obstructive azoospermia who underwent TESE to investigate the expression of spermatogenesis-related genes MND1, SPATA22, GAPDHS and ACR.MethodsTesticular biopsy specimens were subdivided into three groups: hypospermatogenesis (HS); maturation arrest (MA); and Sertoli cell-only syndrome (SCO). The levels of expression of the spermatogenesis-related genes MND1, SPATA22, GAPDHS and ACR in the testes were compared among these three groups using the reverse transcription polymerase chain reaction (RT-PCR) technique.ResultsAnalysis of the expression of spermatogenic genes in human testes with abnormal spermatogenesis showed different expression patterns in patients from different groups. Fertilization rate for studied set of patients was 66% and pregnancy rate 29%. For HS group fertilization rate was 72% and pregnancy rate 32%, while for MA group fertilization and pregnancy rates were 54% and 26%, respectively. Fertilization rates in relation to the studied genes were uniformly around 70%, pregnancy rates for ACR and GAPDHS genes were surprisingly low at 6% and 8% correspondingly.ConclusionsAnalysis of the expression of genes involved in spermatogenesis can be a fast additional test for the level of spermatogenesis in testicular samples.

Microsurgical versus conventional single-biopsy testicular sperm extraction in nonobstructive azoospermia: a prospective controlled study

To compare the success rates of sperm retrieval using either microsurgical testicular sperm extraction (micro-TESE) or conventional testicular sperm extraction (TESE) in a group of men with nonobstructive azoospermia (NOA). Prospective and controlled study. Sixty men with NOA were enrolled in this study and each patient served as his own control. For conventional TESE, two large (6x6x6 mm) biopsies of random testicular parenchyma were obtained. One biopsy specimen was sent for pathology examination and classified according to the predominant histological pattern, i.e., Sertoli-cell only (SCO), maturation arrest (MA) and hypospermatogenesis (HYPO), while the other was dissected and examined for the presence of sperm in the embryology laboratory (EL). After that, the testicular microdissection was carried out as described by Schlegel (1998), with minor modifications. Tubules were sent to the EL for sperm search. Successful sperm retrieval (SSR) rates, both overall and stratified by histology category, were compared between conventional and microsurgical TESE. Overall, the SSR rate was significantly higher using micro-TESE (27/60; 45%) as compared to TESE (15/60; 25%; P=0.02). In successful retrievals, a positive retrieval with TESE was always coupled with a positive one by micro-TESE. SSR rates according to testicular histology results were 92.9% (13/14), 63.6% (7/11) and 20.0% (7/35) in the subgroups of HYPO, MA and SCO, respectively. In addition, micro-TESE success rates were significantly higher than TESE in all testicular histology categories of HYPO (92.9% micro-TESE; 64.3% TESE), MA (63.6% micro-TESE; 9.1% TESE) and SCO (20% micro-TESE; 5.7% TESE) (P<0.01). Micro-TESE is a better method to retrieve spermatozoa from men with NOA than conventional single-biopsy TESE. SSR rates in NOA, either by micro-TESE or TESE, are correlated to the testicular histopathology results. Regardless of testicular histology category, micro-TESE yields higher SSR rates than TESE.

Sperm retrieval and the predictive parameter of non-obstructive azoospermia: a meta-analysis of literatures 1990 to 2008

To investigate which sperm retrieval technique is suitable for the non-obstructive azoospermia (NOA) patient, and to identify the relevant predictive parameters. Literatures on NOA patients who had undergone sperm retrieval and pathological examination of testis were identified from Cochrane Library, CNKI and Medline (1990 to 2008) and analyzed. Twenty-five articles were enrolled. When testicular fine needle aspiration (TEFNA) was compared with testicular sperm extraction (TESE), the sperm retrieval rate of the former was 23.0%, significantly lower than that of the latter (52.2%, RR: 0.49, 95%CI: 0.41 - 0.60, P < 0.05); and when TESE was compared with micro-surgical testicular sperm extraction (mTESE), the sperm retrieval rate of the former was 35.7%, significantly lower than that of the latter (54.6%, RR: 0.70, 95%CI: 0.50 - 0.98, P < 0.05). Sperm retrieval rate was closely correlated with the testicular pathological category of the NOA patients. The sperm retrieval rates of the patients with hypospermatogenesis (HS), maturation arrest (MA), and Sertoli cell only syndrome (SCOS) were 76.7%, 46.2%, and 32.8% respectively (RR: 1.65, 2.40, 1.50; 95%CI: 1.21 - 2.91, 1.85 - 6.90, 1.02 - 2.26, P < 0.05). mTESE is the best sperm retrieval technique. A better to choice before deciding the treatment program of NOA patients is to identify the testicular pathological category in the NOA patients, and then to predict the outcome of TESE before assisted reproduction technology.

Recovery of spermatogenesis after microsurgical subinguinal varicocele repair in azoospermic men based on testicular histology

Analyze whether testicular histologic patterns from a group of azoospermic men with varicocele is predictive of treatment outcome after subinguinal microsurgical varicocele repair. Seventeen azoospermic men underwent bilateral open single testis biopsy and microsurgical subinguinal repair of clinical varicoceles. Histopathology of testicular biopsies revealed hypospermatogenesis (HYPO) in 6 men, maturation arrest (MA) in 5, and Sertoli cell-only (SCO) in 6. Overall, presence of spermatozoa in the ejaculates was achieved in 47% (8/17) of men after varicocele repair, but only 35% (6/17) of them had motile sperm in their ejaculates. Only men with testicular histology revealing HYPO (5/6) or maturation arrest (3/5) had improvement after surgery. Median (25%-75% percentile) postoperative motile sperm count for both groups were 0.9 x 10(6)/mL (0.1-1.8 x 10(6)/mL) and 0.7 x 10(6)/mL (0.1-1.1), respectively (p = 0.87). The mean time for appearance of spermatozoa in the ejaculates was 5 months (3 to 9 months). One (HYPO) of 8 (12.5%) men who improved after surgery contributed to an unassisted pregnancy. Postoperative testicular biopsies obtained from patients who had no improvement after surgery revealed that testicular histology diagnosis remained unchanged. Successful testicular sperm retrieval for intracytoplasmic sperm injection (ICSI) was achieved in 4 of 9 (44.4%) individuals who did not improve after surgery, including 1 man with testicular histology exhibiting SCO. Microsurgical varicocele repair in nonobstructive azoospermic men with clinical varicoceles can result in sperm appearance in the ejaculate when hypospermatogenesis or maturation arrest is found on testicular histology diagnosis.

Efficacy of Cryopreservation of Testicular Sperm in Hypo Spermatogenesis

Testicular sperm using ICSI could achieve optimal fertilization and pregnancy. Hypospermatogenesis is defined as a decrease in the number of spermatogenic cells, so that few mature spermatozoa are formed. This study was performed to assess the fertilizing ability and its embryonic developmental capacity using fresh- and frozen-thawed testicular sperm in hypospermatogenesis patients.