Nunziata Barone

Sperm aneuploidy in infertile men

A recent line of research has shown that infertile male patients produce cytogenetically abnormal spermatozoa, despite a normal somatic karyotype, as a result of an altered intra-testicular environment that affects negatively the mechanisms controlling chromosome segregation during cell division. The rate of aneuploid spermatozoa production is significantly higher in patients with abnormal sperm parameters compared with those of normozoospermic subjects or infertile patients with normal sperm parameters. All chromosomes are subject to aneuploidy, although at a different rate; the heterochromosomes are more often altered than are the autosomes. A negative correlation has been reported to exist between aneuploidy and the main sperm parameters, suggesting that greater testicular damage is associated with a greater chance of chromosome malsegregation events. Abnormally-shaped spermatozoa are more likely to have chromosome abnormalities, particularly those with an enlarged head. More studies are necessary, however, to evaluate whether other types of sperm head abnormalities are also associated with an abnormal sperm chromosome complement. The possibility of retrieving testicular or epididymal spermatozoa in patients with azoospermia and using them in assisted reproduction techniques has prompted the evaluation of their chromosomal status. Studies have shown that testicular and epididymal spermatozoa have a greater rate of aneuploidy compared with that of ejaculated spermatozoa. Some authors have also shown that patients with non-obstructive azoospermia have a significantly higher sperm aneuploidy rate compared with that of patients with obstructive azoospermia. Sperm aneuploidy seems to have a negative impact on assisted reproduction technique outcome. Although it does not affect the fertilization rate, an elevated sperm aneuploidy rate is associated with a greater rate of pregnancy failure. Nevertheless, some patients with elevated sperm aneuploidy rate can still achieve a pregnancy, but with an increased risk of generating an aneuploid offspring. Thus, sperm aneuploidy evaluation is recommended in infertile patients with abnormal semen parameters, particularly if they undergo IVF programmes.

Spontaneous transmission from a father to his son of a Y chromosome microdeletion involving the deleted in azoospermia (DAZ) gene

Microdeletions of the so-called azoospermia factor (AZF) locus of the Y chromosome long arm (Yq) have been recognized as an etiological factor of severe oligozoospermia or azoospermia. Because of this, patients affected are generally infertile unless assisted reproductive techniques are used. We report the case of an oligozoospermic patient (proband) who inherited an extensive Yq microdeletion from his father through a spontaneous pregnancy. Leukocyte DNA was extracted using a commercially available kit. A total of 22 pairs of sequence-tagged site (STSs) based primers, spanning the entire AZF region, were used for the screening. Both the proband and his father carried a Yq microdeletion of the most distal AZF subregion (AZFc) where the deleted in azoospermia (DAZ) gene is located. The proband’s father was a sixty-nine-yr-old man who had 3 other children, 2 females and 1 male. This case adds further evidence that the deletion of the DAZ gene is associated with different phenotypic expressions, including normal fertility.

Chromosome analysis of epididymal and testicular spermatozoa in patients with azoospermia.

Azoospermic patients can now father children once spermatozoa have been retrieved from the epididymis or the testis. However, there are concerns about the risk of chromosomal abnormalities since an increase in sperm aneuploidy rate has been reported in samples from patients with abnormal sperm parameters. The purpose of this study was therefore to evaluate the sperm aneuploidy and diploidy rates for chromosomes 8, 12, 18, X and Y in spermatozoa extracted from the epididymes (n=10) or the testes (n=6) of patients with azoospermia. Ejaculated spermatozoa of healthy men (n=14) served as control. Epididymal and testicular spermatozoa had an aneuploidy rate significantly higher than that found in ejaculated spermatozoa. The aneuploidy and diploidy rates of testicular spermatozoa were higher, but not significantly different, than those found in epididymal spermatozoa. This study has shown that azoospermic patients have an increased sperm aneuploidy rate. They should therefore be given appropriate genetic counselling before entering in-vitro fertilisation programs.

Structure of spermatodesms of Orthoptera Tettigonioidea

Examination of spermatodesms collected from the male and female genital tracts of numerous Orthoptera Tettigonioidea revealed an overall morphological and ultrastructural organization that is generally similar in individuals of the same sex but considerably different between males and females of even the same species. In the male genital tracts each spermatodesm is composed of a limited number of spermatozoa whose nuclei and acrosomes are covered by a mucous cap. The spermatozoa inside each bundle are mainly arranged in parallel rows and are always distinctly separate. The number of spermatozoa per spermatodesm may vary within the same individual although it does not seem to exceed a maximum value that we could only determine exactly in Tettigoniidae species. The most characteristic feature of spermatozoa of all the species examined is a conspicuous elongation of the plasma membrane in the acrosomal region that is not present in the female genital tracts. In addition, spermatodesms from females are composed of highly numerous tightly packed spermatozoa that are linked together via the acrosomal region. This characteristic of spermatodesms, never previously reported in other insect species, would involve their reorganization during transfer from the male to the female genital tracts and would seem to take place in the spermatophore. The probable role of spermatodesms in the reproductive physiology of Tettigonioidea might be related to the degree of maturity of the sex cells transferred to the female; the reorganization of the spermatozoa out of the male genital tracts seems to support this hypothesis.

Follicle-stimulating hormone treatment in normogonadotropic infertile men

Several empirical treatments have been proposed to treat idiopathic infertility in men, including follicle-stimulating hormone (FSH). FSH administration is effective in patients with hypogonadotropic hypogonadism, which suggests it might be useful in patients with oligozoospermia who have normal FSH levels. Indeed, many studies have evaluated the efficacy of FSH administration in these patients, several of which have shown improvements in sperm parameters. By contrast, other studies have not reported any significant effect of FSH administration on conventional sperm parameters, although some of have reported the normalization of spermatozoon ultrastructural morphology, as well as reductions in DNA fragmentation, production of reactive oxygen species and aneuploidy. Contemporary studies suggest that the response to FSH treatment in oligozoospermic patients might, at least partially, reflect polymorphisms of the FSH receptor gene. Thus, FSH administration in oligozoospermic men with normal serum FSH levels might be efficacious only in selected patients. For this reason, additional studies are needed to determine the predictive factors and clinical conditions that can be used to identify patients who could benefit from FSH treatment.

Spermiolytic activity of the epithelium of the spermathecal duct of Rhacocleis annulata fieber (Orthoptera : Tettigoniidae)

Abstract This study reports, for the first time, lytic activity in the distal and median regions of the spermathecal duct of Rhacocleis annulata Fieber (Orthoptera: Tettigoniidae). Lysis is more pronounced in recently mated females, and appears to take place in a progressive sequence originating in the cells underlying the cuticular intima. These cells exhibit large heterogeneous vesicles resembling lysosomes, which would participate in the partial dismantling of the cuticular intima. Consequently, the sperm found in the lumen of the latter organ penetrate the cells, probably pushed by the pressure exerted on the luminal walls, and then, together with the cells themselves, undergo lysis. Degradation of spermatozoa seems to follow a strict sequence of events, which ends with the dismantling of the microtubules of the axoneme, as reported previously for Locusta (Cantacuzene, 1971) and for some pulmonate gastropods (Bayne, 1970). We propose that lysis is part of the mechanism dealing with capture and subsequent elimination of surplus sperm transferred to the female during copulation. Accordingly, fertilization activity would occur exclusively within the seminal receptacle. No evidence of a similar phenomenon in the latter organ has previously been reported for Rhacocleis annulata.

PARP-1 and CASP3 genes are up-regulated in LNCaP and PC-3 prostate cancer cell lines

Prostate cancers (PCa) are slow growing; however, some are more aggressive [1]. Men who have a first-degree relative (father or brother) with PCa have twice the risk of developing PCa, and those with two first-degree relatives affected have a fivefold greater risk compared with men with no family history [2, 3]. Many factors, including genetics, habit and diet, have been implicated in the development of PCa [4]. The genetic background may contribute to prostate cancer risk, as suggested by associations with race, family, and specific gene variants. In the United States, PCa more commonly affects black men than white or Hispanic men; in addition, it is also more deadly in black men [5]. In contrast, the incidence and mortality rates for Hispanic men are one-third lower than for non-Hispanic whites. Studies of twins in Scandinavia suggest that 40 % of PCa risk can be explained by inherited factors [6]. Cancer cells may metastasize from the prostate to other parts of the body, particularly the bones and lymph nodes. PCa may cause pain, difficulty in urinating, problems during sexual intercourse, or erectile dysfunction [7]. Moreover, there is a significant number of men who require systemic therapy and ongoing surveillance for advanced PCa [8]. High throughput technologies, such as DNA and protein microarrays, have enabled the identification of genes (some associated with apoptosis) and their corresponding proteins that are differentially regulated in malignant conditions [9]. Inhibition of apoptosis is a critical pathophysiological factor that contributes to the onset and progression of PCa, but the molecular mechanisms are not entirely understood. Therefore, insight into the mechanism(s) of the misregulation of apoptosis could be the basis for developing more effective therapeutic approaches to destroy apoptosisresistant tumor cells, as found in PCa [10]. Apoptosis is a programmed cell death process that takes place under normal physiological and pathological conditions. Caspases, a family of cysteine-dependent aspartatespecific proteases, are important mediators of the apoptotic process [11], Among them, caspase 3 (CASP3; OMIM 600636), which maps to human chromosome 4q35, is involved in both extrinsic and intrinsic apoptotic pathways. CASP3 is a member of the caspase family of cysteine proteases which is central to the cell death pathway, as the extrinsic and intrinsic cell death pathways converge to activate this protein for the final execution of apoptosis [12]. Activation of PARP-1 gene (OMIM 173870), located to 1q42, is required for translocation of the apoptosisinducing factor (AIF) from the mitochondria to the nucleus, and it is proteolytically cleaved at the onset of apoptosis by CASP3 [13]. Furthermore, PARP-1 activity and poly (ADP-ribose) (PAR) polymer mediate PARP1-induced cell death such as apoptosis or macroautophagocytotic cell death [13]. Overall, the most striking differences in PARP-1 gene expression have been observed in breast, ovarian, endometrial, lung, skin cancers, and non-Hodgkin’s lymphoma [14]. M. Salemi (&) IRCCS Associazione Oasi Maria SS Institute for Research on Mental Retardation and Brain Aging, Troina, Italy e-mail: micezia@tiscali.it