Sperm Chromosome Research Articles

Association, prevalence, and clearance of human papillomavirus and antisperm antibodies in infected semen samples from infertile patients

To evaluate prevalence, association, and clearance of human papillomavirus (HPV) and antisperm antibodies (ASAs) in infected semen samples from infertile patients. Cross-sectional clinical study. Andrology and microbiology sections at a university hospital. Three groups of subjects: 61 infertile patients with HPV semen infection, 104 noninfected infertile subjects, and 92 control subjects. Semen analysis, spermMar test, fluorescence in situ hybridization for sperm aneuploidy and for HPV, and immunofluorescence for HPV 16-L1 and immunoglobulins (IgA, IgG, and IgM) determination. Association of sperm procedures, HPV sperm infection, sperm aneuploidies, and sperm ASAs. Infertile patients with HPV semen infection showed high percentages of ASAs. In these patients HPV sperm infection was associated with lower sperm motility, which was worse in subjects with ASAs. No alterations of sperm chromosomes were observed. To obtain a significant clearance of both HPV sperm infection and ASAs at least 24 months of follow-up were needed. Human papillomavirus has been recently suggested to have an important role in male infertility. This study demonstrated that HPV sperm infection can be long lasting and frequently associated with ASAs that may further reduce male fertility. Infertile patients with positive spermMar test results should be considered for investigation for HPV, especially if they are candidates for assisted reproduction.

Hierarchical radial and polar organisation of chromosomes in human sperm

It is well established that chromosomes occupy distinct positions within the interphase nuclei, conferring a potential functional implication to the genome. In addition, alterations in the nuclear organisation patterns have been associated with disease phenotypes (e.g. cancer or laminopathies). The human sperm is the smallest cell in the body with specific DNA packaging and the mission of delivering the paternal genome to the oocyte during fertilisation. Studies of nuclear organisation in the sperm have postulated nonrandom chromosome position and have proposed a chromocentre model with the centromeres facing toward the interior and the telomeres toward the periphery of the nucleus. Most studies have assessed the nuclear address in the sperm longitudinally predominantly using centromeric or telomeric probes and to a lesser extent with whole chromosome paints. To date, studies investigating the radial organisation of human sperm have been limited. The purpose of this study was to utilise whole chromosome paints for six clinically important chromosomes (18, 19, 21, 22, X, and Y) to investigate nuclear address by assessing their radial and longitudinal nuclear organisation. A total of 10,800 sperm were analysed in nine normozoospermic individuals. The results have shown nonrandom chromosome position for all chromosomes using both methods of analysis. We present novel radial and polar analysis of chromosome territory localization within the human sperm nucleus. Specifically, a hierarchical organisation was observed radially with chromosomes organised from the interior to the periphery (chromosomes 22, 21, Y, X, 19, and 18 respectively) and polar organisation from the sperm head to tail (chromosomes X, 19, Y, 22, 21, and 18, respectively). We provide evidence of defined nuclear organisation in the human sperm and discuss the function of organisation and potential possible clinical ramifications of these results in regards to male infertility and early human development.

Risk evaluation and preimplantation genetic diagnosis in an infertile man with an unbalanced translocation t(10;15) resulting in a healthy baby

Balanced chromosome structural rearrangement, including reciprocal translocation, Robertsonian translocation and inversion, is an important cause of male infertility that results in azoospermia, oligozoospermia or asthenozoospermia [1]. Unbalanced translocation between two autosomes, which usually results in mental retardation or multiple congenital anomalies, is rare in infertile patients. To our knowledge, only the t(15;18) unbalanced translocation has been reported, in two healthy infertile brothers. Although fluorescence in situ hybridization (FISH) analysis showed that chromosomal normal sperms existed in their semen, these two brothers and their partners chose donor insemination rather than preimplantation genetic diagnosis (PGD) [2]. Whether patients with this kind of rare karyotype could deliver a healthy baby is unclear. Here, we report a man with severe oligoasthenoteratozoospermia carrying a derivative chromosome 10 resulting from the rare unbalanced translocation t(10;15). Based on risk evaluation by sperm FISH, we performed PGD for the couple and successfully established a normal pregnancy. Our results show that patients with such an abnormal karyotype can produce their own offspring, and emphasize the importance of risk evaluation using sperm FISH for infertile male patients. Patient data A couple presented to our reproduction center due to primary infertility that had persisted for 5 years. The woman showed no obvious cause of infertility on examinations, which included: (1) clinical findings: physical examination and medical history evaluation,such as menstrual history, obstetrical history, sexual history, family history, personal and lifestyle history; (2) biochemical tests: liver and renal function; blood cholesterol, blood triglycerides and blood glucose; and immunologic function; and (3) hormonal assay: estradiol, testosterone, progesterone, follicle stimulating hormone (FSH), luteinizing hormone, prolactin, 17-hydroxypregnenolone and thyroid stimulating hormone (TSH); (4) other diagnosis test such as hysterosalpingography. Her karyotype was also normal. However, the man was found to have severe oligoasthenoteratozoospermia after three routine sperm analyses and one sperm morphology examination following the WHO guidelines (Table 1). Conventional G-banding (G-bands after trypsin and Giemsa, GTG) analysis showed that he had only 45 chromosomes, with one derivative chromosome comprising chromosomes 10 and 15 (Fig. 1). Table 1 Semen analysis Fig. 1 a Partial karyotype of the patient showing normal chromosome 10,15 and der(10). b ideogram of the normal chromosome 10,15 and rearranged chromosome. Red, green and aqua bands show the position and color of the FISH probes used for preimplantation genetic ... The couple then accepted a genetic counseling and asked for further fertility evaluation. Oligoasthenoteratozoospermia might have been the cause of the primary infertility in this couple. The husband carried a rare unbalanced translocation comprising chromosome 10 and chromosome 15 that had not been reported previously. The genetic risk was unclear for this kind of abnormal karyotype. His chromosomal structure was similar to that of a Robertsonian translocation; therefore, the meiosis pattern might also be similar to a Robertsonian translocation. Sperm chromosome constitution can be obtained by sperm FISH analysis and might predict the probability of producing an embryo with normal karyotype. If chromosomally normal sperms are present in semen at a high prevalence, PGD can be performed to select embryos with balanced chromosomes for transfer and deliver a healthy baby. This study was approved by the Institutional Review Board of Central South University and Reproductive and Genetics Hospital of Citic-Xiangya, and the patients’ written informed consent was obtained.

Exploring the relationship between the severity of oligozoospermia and the frequencies of sperm chromosome aneuploidies

The study was aimed to investigate the association between the degree of oligozoospermia and sperm chromosome aneuploidy frequencies in male infertility and to determine whether chromosomal profiles of sperm nuclei would be used for a supportive test before additive reproduction technics. The meiotic segregation profiles of chromosomes X, Y, 13, 18 and 21 were compared by fluorescent in-situ hybridisation (FISH) on the spermatozoa of 30 normally karyotyped oligozoospermic (10 mild, 11 moderate, nine severe) cases without Y-microdeletions, and 10 normozoospermic cases. The results showed significantly higher frequencies of chromosomes 13, 18, 21 disomies (P<0.001) in the group of patients with moderate and severe oligozoospermia compared with the disomy frequencies of normozoospermic group. The statistically significant differences were also determined in disomy frequencies of sex chromosomes (XY, XX and YY) in between oligozoospermic and normozoospermic groups (P<0.001, P<0.001, P<0.040, respectively). Because oligozoospermic patients are the ones consulted the most for assisted reproductive techniques, identification of sperm aneuploidy rates in men could be considered as an appropriate supportive test before the reproductive implementations. Furthermore, the patients should be counselled with respect to genetic screening results for the potential risk of aneuploid embryo and pre-implantation genetic diagnosis or prenatal diagnosis.

Comparison of aneuploidies of chromosomes 21, X, and Y in the blood lymphocytes and sperm of workers exposed to benzene

Benzene is a primary industrial chemical and a ubiquitous environmental pollutant that causes human leukemia and maybe other malignancies. Occupational exposure to benzene has been associated with increased chromosomal aneuploidies in blood lymphocytes and, in separate studies, in sperm. However, aneuploidy detection in somatic and germ cells within the same benzene-exposed individuals has never been reported. To compare aneuploidies in blood lymphocytes and sperm within the same individuals exposed to benzene, a cross-sectional study was conducted in 33 benzene-exposed male workers and 33 unexposed workers from Chinese factories. Air benzene concentrations in the exposed workers ranged from below the detection limit to 24 ppm (median, 2.9 ppm) and were undetectable in the unexposed subjects. Aneuploidies of chromosomes 21, X, and Y in blood lymphocytes were examined by multicolor fluorescence in situ hybridization and were compared to the previously reported aneuploidies in sperm. The results showed that benzene exposure was positively associated with the gain of chromosome 21 but not sex chromosomes in blood lymphocytes. This was in contrast to analysis of sperm, where the gain of sex chromosomes, but not chromosome 21, was significantly increased in the exposed workers. Furthermore, a significant correlation in the gain of sex chromosomes between blood lymphocytes and sperm was observed among the unexposed subjects, but not among the exposed workers. The findings suggest that benzene exposure induces aneuploidies in both blood cells and sperm within the same individuals, but selectively affects chromosome 21 in blood lymphocytes and the sex chromosomes in sperm.

The Integrated HIV-1 Provirus in Patient Sperm Chromosome and Its Transfer into the Early Embryo by Fertilization

Complete understanding of the route of HIV-1 transmission is an important prerequisite for curbing the HIV/AIDS pandemic. So far, the known routes of HIV-1 transmission include sexual contact, needle sharing, puncture, transfusion and mother-to-child transmission. Whether HIV can be vertically transmitted from human sperm to embryo by fertilization is largely undetermined. Direct research on embryo derived from infected human sperm and healthy human ova have been difficult because of ethical issues and problems in the collection of ova. However, the use of inter-specific in vitro fertilization (IVF) between human sperm and hamster ova can avoid both of these problems. Combined with molecular, cytogenetical and immunological techniques such as the preparation of human sperm chromosomes, fluorescent in situ hybridization (FISH), and immunofluorescence assay (IFA), this study mainly explored whether any integrated HIV provirus were present in the chromosomes of infected patients' sperm, and whether that provirus could be transferred into early embryos by fertilization and maintain its function of replication and expression. Evidence showed that HIV-1 nucleic acid was present in the spermatozoa of HIV/AIDS patients, that HIV-1 provirus is present on the patient sperm chromosome, that the integrated provirus could be transferred into early embryo chromosomally integrated by fertilization, and that it could replicate alongside the embryonic genome and subsequently express its protein in the embryo. These findings indicate the possibility of vertical transmission of HIV-1 from the sperm genome to the embryonic genome by fertilization. This study also offers a platform for the research into this new mode of transmission for other viruses, especially sexually transmitted viruses.

Meiotic segregation and interchromosomal effects in a rare (1:2:10) complex chromosomal rearrangement

Complex chromosomal rearrangements (CCR) are rare rearrangements involving more than two chromosomes and more than two breakpoints. CCR are associated with male infertility as a result of the disruption of spermatogenesis due to complex meiotic configurations and the production of chromosomally abnormal sperm. We examined a carrier of a t(1:2:10) CCR in order to determine the patterns of segregation and any presence of an interchromosomal effect (ICE). Centromeric, locus specific and telomeric probes (Vysis, USA) were used for the study. On ~1,000 sperm nuclei from the reciprocal translocation carrier, dual color Fluorescence in situ hybridization (FISH) was performed on each of the involved chromosomes to determine the patterns of segregation. FISH was also performed on chromosome 13, 18, 21, X and Y to determine any ICE. We observed abnormal chromosome complements in 24.3%, 19.5% and 15.8% of sperm for chromosomes 2, 10 and 1, respectively. There was a significantly increased rate of ICEs for chromosomes 13 and 21 when compared with controls. CCR may present a lower risk for producing unbalanced chromosomes than other studies have indicated. CCRs may be at an increased risk for ICE especially among acrocentric chromosomes.

Mitotic chromosome size scaling in Xenopus

As rapid divisions without growth generate progressively smaller cells within an embryo, mitotic chromosomes must also decrease in size to permit their proper segregation, but this scaling phenomenon is poorly understood. We demonstrated previously that nuclear and spindle size scale between egg extracts of the related frog species Xenopus tropicalis and Xenopus laevis, but show here that dimensions of isolated mitotic sperm chromosomes do not differ. This is consistent with the hypothesis that chromosome scaling does not occur in early embryonic development when cell and spindles sizes are large and anaphase B segregates chromosomes long distances. To recapitulate chromosome scaling during development, we combined nuclei isolated from different stage Xenopus laevis embryos with metaphase-arrested egg extracts. Mitotic chromosomes derived from nuclei of cleaving embryos through the blastula stage were similar in size to replicated sperm chromosomes, but decreased in area approximately 50% by the neurula stage, reproducing the trend in size changes observed in fixed embryos. Allowing G2 nuclei to swell in interphase prior to mitotic condensation did not increase mitotic chromosome size, but progression through a full cell cycle in egg extract did, suggesting that epigenetic mechanisms determining chromosome size can be altered during DNA replication. Comparison of different sized mitotic chromosomes assembled in vitro provides a tractable system to elucidate underlying molecular mechanisms.

Phylogeny and classification of Pholcidae (Araneae): an update

For over 100 years, Eugene Simon's system of pholcid classification has been used with only minor modifications. Phylogenetic research over the last decade has shown that some fundamental changes are necessary if the formal system is to reflect putative evolutionary relationships. Based on cladistic analyses of morphological and molecular data and on qualitative character assessment, the family is here divided into five subfamilies: Ninetinae, Arteminae, Modisiminae, Smeringopinae, and Pholcinae. All currently valid genera are placed in a cladogram. Even though the evidence supporting some of the nodes and assignments is weak, the cladogram generates numerous testable hypotheses and provides an improved framework for the mapping of 'new' characters like those from sperm ultrastructure and chromosome analysis.

Aneuploidy frequency in sperm of fertile men

Analysis of sperm aneupoidy in 11 healthy men using two- or three-color FISH permitted to determine the average frequency of disomy in chromosomes 13 and 21 (0.11% and 0.2%, respectively), disomy in chromosome 18 (0.05%) and reveal gonosomal aneuploidy variants and their frequency. The frequency of XX disomy was 0.04%; XY, 0.17%; YY, 0.06%; and gonosomal nullisomy, 0.29%. We also assessed the frequency of meiotic nondisjunction of chromosomes 13, 21, 18, X, and Y and the frequency of XX, XY, and YY diploid chromosomes in sperm. The XY variant prevailed in gonosomal aneuploidy and diploidy and was associated with abnormal chromosomal segregation in meiotic anaphase I. The contribution of human sperm chromosomal imbalance to early embryonic lethality and to some chromosomal diseases of syndrome type in the offspring is discussed.

Semi‐automated scoring of triple‐probe FISH in human sperm: Methods and further validation

Although the frequency and consequence of sperm chromosomal abnormalities are considerable, few epidemiologic studies in large samples have been conducted to investigate etiologic risk factors. This is, in part, attributable to the labor intensive demands of manual sperm fluorescence in situ hybridization (FISH) scoring. As part of an epidemiologic study investigating environmental risk factors for aneuploidy among men attending a hospital-based fertility clinic, a semi-automated method of slide scoring was further validated and used to estimate sex chromosome sperm disomy frequency in a large number of samples. Multiprobe FISH for chromosomes X, Y, and 18 was used to determine sex chromosome disomy in sperm nuclei. Semi-automated scoring methods were used to quantify X disomy (sperm FISH genotype XX18), Y disomy (YY18), and XY disomy (XY18). The semi-automated results were compared with the results from manual scoring in 10 slides. The semi-automated method was then used to estimate sex chromosome disomy frequency in 60 men. Of 10 slides scored, significant differences between the manual and semi-automated results were seen primarily in one slide that was of poor quality because of over swollen nuclei. Among 60 men analyzed using the semi-automated method, median total sex chromosome disomy frequency was 1.65%, which is higher than seen among normal men but within range with reports from fertility clinic populations. These results further validate that semi-automated methods can be used to score sperm disomy with results comparable to manual methods. This is the largest study to date to provide estimates of sex chromosome disomy among men attending fertility clinics. These methods should be replicated in larger clinic populations to arrive at stable estimates of aneuploidy frequency in men who are members of subfertile couples. © 2011 International Society for Advancement of Cytometry.

An investigation of the potential effect of vacuoles in human sperm on DNA damage using a chromosome assay and the TUNEL assay

The aims of this study were to establish whether individual differences exist in the frequency and size of vacuoles found in human sperm and to ascertain whether such vacuoles are involved in causing DNA damage. Morphologically normal sperm were obtained from 15 IVF and 2 ICSI patients and 3 fertile donors. (i) Sperm heads were analyzed for the presence of vacuoles under a 1000× differential interference contrast microscope. (ii) In three patients and two donor samples, structural chromosomal damage was evaluated in normal sperm containing large vacuoles and selected at 1000× magnification for injection into mouse oocytes. (iii) In 10 patients and two donor samples, confocal laser microscopy detected DNA damage in sperm-exhibiting large vacuoles and stained with an in situ cell death detection kit. (i) Vacuoles were observed in almost all normal sperm from patient and donor ejaculates and were mainly located at the tip or middle area of the sperm heads. However, average incidence of normal sperm exhibiting large vacuoles was 4.6 and 4.2% in the patient and donor groups, respectively. (ii) Sperm chromosome assays did not reveal any differences in the incidence of structural chromosome aberrations between sperm exhibiting large vacuoles and those without them (9.1 versus 4.1%). (iii) No significant difference in frequency of TUNEL-positive cells was found between normal sperm with large vacuoles and those without them in the samples examined. Among 227 sperm exhibiting large vacuoles, only 7 cells were TUNEL positive. The results showed that large vacuoles were not responsible for DNA damage, suggesting that intra-cytoplasmic injection of morphologically selected sperm may not be required for patients who produce high-quality semen.

Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction

The objective of this study was to determine if a relationship exists between sperm parameters, measured by sperm chromatin structure assay (SCSA), and spontaneous abortion and multiple births in couples undergoing assisted reproduction treatment. Retrospective analysis of infertility treatment outcomes and occurrence of spontaneous abortion and multiple births was conducted in 233 couples who underwent treatment by intracytoplasmic sperm injection or intrauterine insemination at the Sher Institute for Reproductive Medicine, between 2001 and 2004. Sperm samples used for treatments were analysed for sperm concentration, sperm motility and two different parameters of SCSA (DNA fragmentation index, DFI, and high DNA stainability, HDS). Pregnancy, spontaneous abortion and multiple birth rates were recorded for all couples. A statistically significant relationship ( P < 0.001) was observed between DFI and spontaneous abortion. However, the correlation between HDS and spontaneous abortion was not statistically significant. Significantly lower levels of DFI were observed in men from couples having triplet pregnancies compared with those in the spontaneous abortion group ( P ⩽ 0.05). It is concluded that the parameters of SCSA correlate significantly with spontaneous abortion and multiple birth and may provide guidance for clinical decision making (number of embryos per transfer) and management of spontaneous abortion-prone cases. The goal of this study was to determine if a relationship exists between the sperm chromatin structure assay (SCSA) and spontaneous abortion and multiple births in couples treated via assisted reproductive technologies such as intra-cytoplasmic sperm injection or intrauterine insemination. SCSA is a test used by fertility clinics to determine if the sperm chromosomes or DNA are properly packaged in the sperm head and are therefore protected from damage that could occur during travel in the female reproductive system. A retrospective analysis of the results of SCSA, infertility treatment outcomes and the occurrence of spontaneous abortions and multiple births was conducted for 233 couples attending an infertility clinic. A statistically significant correlation was found between one of the SCSA parameters and spontaneous abortion. In addition, lower levels of this SCSA parameter were observed in men from couples having triplet pregnancies. We conclude that the parameters of SCSA correlate significantly with spontaneous abortions and multiple births and may provide guidance for clinical decision making, such as number of embryos per transfer or intensive management of spontaneous abortion-prone cases.

Spermatogenesis in Klinefelter syndrome

Klinefelter syndrome (KS) (47,XXY) is the most common sex chromosomal disorder, and it is a frequent form of male hypogonadism and infertility. Although the majority of these patients are azoospermic, they might have severe oligozoospermia or residual single-residual foci with spermatogenesis in the testis. We report our experience on sperm retrieval in the ejaculate and testis, and evaluate the frequency of chromosome abnormalities in sperm of KS. Eighty-four 47,XXY KS were evaluated with seminal analysis, body hair distribution, reproductive hormones, ultrasonographic scanning of the testis and prostate, bilateral testicular sperm extraction (TESE), sperm or testicular cells sex chromosomes aneuploidies. Out of 84 patients, 7 (7/84; 8.3%) had sperm in the ejaculate. Out of the 77 azoospermic patients, 24 underwent TESE and 9 (9/24; 37.5%) had successful sperm recovery. The comparison of reproductive hormones, age and testicular volume did not show significant differences between patients with and without successful sperm recovery in semen or TESE . Patients without successful sperm recovery in semen analysis or TESE had signs of hypoandrogenism more evident than patients with successful sperm recovery. Patients with KS produced a higher number of sperm aneuploidy with respect to normozoospermic fertile controls and non-genetic severely oligozoospermic men. Men with KS are not always sterile. In some of these patients sperm can be found in semen or in the testis, but the proportion of sperm aneuploidy is high. Signs of hypoandrogenism seem to be associated with low sperm recovery rate.

Sperm DNA in Grasshoppers: Structural Features and Fertility Implications

Abstract This contribution briefly reviews the importance of grasshopper chromosomes in evolutionary studies and in achieving a comprehensive view of cytogenetics, emphasizing that meiotic chromosomes have been the traditional means for analyzing the nature of such problems. Structural changes occurring in the sperm, as a direct consequence of some chromosomal changes, have received less attention. We propose that grasshopper sperm could be a good model for the study of: 1) aspects of genome territoriality, and 2) consequences in the sperm DNA molecule, derived from a compromised meiotic process, linked with naturally forced genome hybridization. The results presented here, by using appropriate DNA probes for genome regionalization, allow the proposal of a grasshopper sperm chromosome organization. Centromere regions are distributed and concentrated at the proximal end of the sperm head where the flagellum is inserted. Also, chromosome arms are longitudinally organized from the proximal to the distal end,...

Higher Proportion of Haploid Round Spermatids and Spermatogenic Disomy Rate in Relation to Idiopathic Male Infertility

To investigate the possible association between the spermatogenic chromosomal aberrations and idiopathic male infertility with normal semen volume, sperm concentration, sperm number per ejaculum, and sperm motility. Male infertility can be considered as a syndrome that results from many congenital or acquired illness. Currently, there is an increasing awareness of the potential role of chromosomal factors in the idiopathic forms of male infertility. A total of 202 idiopathic infertile men and 41 normal fertile donors were recruited into this study, through eligibility screening procedures. Proportions and chromatin status of sperm were assessed by flow cytometry after staining the DNA with propidium iodide. Sperm chromosome aneuploidy rates for chromosome 13, 18, 21, X, and Y were evaluated by fluorescence in situ hybridization using five-color probes. Infertile males showed a significantly lower percentage of mature haploid sperm cells and higher percentage of haploid round spermatids than the controls. A significantly higher frequency of disomy on spermatozoa was also detected in infertile males compared with the controls. These findings suggest that higher frequencies of haploid round spermatids and spermatogenic disomy rate may contribute to elevated risk of idiopathic male infertility.

Advanced age increases chromosome structural abnormalities in human spermatozoa

This study explores the relationship between sperm structural aberrations and age by using a multicolor multichromosome FISH strategy that provides information on the incidence of duplications and deletions on all the autosomes. ToTelvysion kit (Abbott Molecular, Abbott Park, IL, USA) with telomere-specific probes was used. We investigated the sperm of 10 male donors aged from 23 to 74 years old. The donors were divided into two groups according to age, a cohort of five individuals younger than 40 and a cohort of five individuals older than 60 years. The goal of this study was to determine (1) the relationship between donor age and frequency and type of chromosome structural abnormalities and (2) chromosomes more frequently involved in sperm structural aberrations. We found that the older patients had a higher rate of structural abnormalities (6.6%) compared with the younger cohort (4.9%). Although both duplications and deletions were seen more frequently in older men, our findings demonstrate the presence of an excess of duplications versus deletions in both groups at a ratio of 2 to 1. We demonstrate that the distribution of duplications and deletions was not linear along the chromosomes, although a trend toward a higher rate of abnormalities in larger chromosomes was observed. This work is the first study addressing the frequencies of sperm chromosome structural aberrations of all autosomes in a single assay thus making a contribution to the clarification of the amount and origin of damage present in human spermatozoa and in relation to age.

Sperm chromosome analysis and preimplantation genetic diagnos is in an infertile male with mosaic trisomy 18

To analyze the numerical aberration rate of X, Y and chromosome 18 in sperms from an oligozoospermic male with mosaic trisomy 18 and to perform preimplantation genetic diagnosis (PGD) for the couple. G-banding and fluorescence in situ hybridization (FISH) were performed on metaphase chromosome. Sperm was analyzed in three-color FISH with a probe mixture containing CEP18, CEPY and Tel Xq/Yq. A healthy man with normal semen parameters was used as control. Significant difference in the rates of disomy for chromosome 18 (0.63% vs. 0.16%) and the gonosomes (0.945% vs. 0.35%) and diploidy (0.87% vs. 0.31%) was found in the spermatozoa between the patient and the control. After four embryos were biopsied in one PGD cycle, two embryos with XY1818 and XX1818 were selected for implanting and clinical pregnancy was ongoing. Sperm-FISH allows further understanding of aneuploidy rate and accurate genetic counseling. FISHPGD was effective for patient with mosaic trisomy 18.

Effects of anti-neoplastic treatment on sperm aneuploidy rate in patients with testicular tumor: A longitudinal study

The adjuvant radio/chemotherapy, usually employed after orchidectomy in patients with testicular tumors, allows a long-term survival with a consequent increased request for fertility. However, little is known about the effects of the anti-neoplastic treatment on sperm cytogenetic asset. Therefore, this prospective, longitudinal study was designed to evaluate the effects of radio- and/or chemotherapy on sperm chromosome. Eleven patients with testicular tumor were enrolled and underwent sperm aneuploidy rate evaluation before and after 3, 6, 9, 12, 18, 24, and 36 months from radio- and/or chemo-therapy ending. A double and triple multicolor fluorescence in-situ hybridizations for chromosomes 8, 12, 18, X and Y were used to evaluate the sperm aneuploidy rate. To define normal sperm aneuploidy rate, 18 healthy, normozoospermic men were selected as controls. Before treatment, testicular tumor patients had a higher total sperm aneuploidy rate compared with normal men. Total sperm aneuploidy rate showed a slight, but statistically significant increase 6 months after anti-neoplastic treatment. This increase was mainly related to the high sperm aneuploidy rate found in 2 patients which remained elevated up to 12 months in both of them. These results showed that anti-neoplastic treatment caused only slight and transient sperm malsegregation events in patients with testicular tumor. However, since a subset of them had an elevated sperm aneuploidy rate for about 1 yr, we suggest to counsel them to refrain from fatherhood for this length of time.

The use of fluorescent in situ hybridization in male infertility

Male factors are implicated in up to 50% of couples being evaluated and treated for infertility with advanced assisted reproductive technologies. Genetic abnormalities, including sperm chromosome aneuploidy as well as structural aberrations, are one of the major causes of infertility. The use of chromosome-specific DNA probes labeled with fluorochromes, particularly the combination with multiple probes, has been used to indirectly study the sperm chromosome by fluorescent in situ hybridization (FISH). Clinically, this technique is also used to assess the sperm of men recovering from gonadotoxic treatment. Recent advances in this technology facilitate the evaluation of sperm aneuploidy. Sperm FISH is a widely used screening tool to aid in counseling couples with severe male factor infertility, especially in cases of prior repeated in vitro fertilization/intracytoplasmic sperm injection failure or recurrent pregnancy loss. Automation of FISH imaging and analysis, as well as the development of emerging techniques such as comparative genomic hybridization, will all contribute to the promise of future diagnostic approaches aimed at improving the quality, ease, and efficiency of aneuploidy analysis.