Study of apoptotic DNA fragmentation in human spermatozoa

Purpose The present study was undertaken to evaluate the effects of morphokinetic abnormalities of human spermatozoa on chromatin packing and DNA integrity and possible beneficial effects of sperm selection in ICSI. Methods Semen samples from 1002 patients were analysed for morphology and motility using CASA. Protamine status and DNA fragmentation were analysed by chromomycin A3 staining and sperm chromatin dispersion assay respectively. Results Sperms with elongated, thin, round, pyri, amorphous, micro and macro forms were significantly higher in teratozoospermic and oligoasthenoteratozoospermic groups. Significant difference in chromatin packing and DNA fragmentation index was observed in these abnormal groups compared with normal. Similarly significant correlation was also seen between abnormal motility parameters and DNA fragmentation index in asthenozoospermic group compared with normal. Conclusions Specific abnormal morphological forms have higher incidence of chromatin packing abnormalities and DNA fragmentation. Using these sperms in ICSI might have an impact on fertilization, embryo development and abortion rates. These can be selectively avoided during ICSI procedure to improve ART outcome.

Section: Discussionsupporting

confidence: 58%

CASA derived human sperm abnormalities: correlation with chromatin packing and DNA fragmentation

Sivanarayana

Krishna

Prakash³

et al. 2012

“…Apoptotic markers like active caspase-3 [1,20,32], phosphatidylserine (PS) externalization [3,31] and DNA fragmentation [6,9,20,21] were described to be present in ejaculated sperm from men with normal semen parameters. However, increased rates of ejaculated sperm apoptosis were described in conditions of abnormal sperm output, mainly related to abnormalities of sperm concentration, morphology or rapid progressive motility values [1,3,6,9,20,21,24,[31][32][33][34].…”

Section: Introductionmentioning

confidence: 99%

Caspase signalling pathways in human spermatogenesis

Almeida

Correia

Rocha

et al. 2013

Purpose Little is known about the apoptotic mechanisms involved in abnormal spermatogenesis. In order to describe the significance of apoptosis in azoospermia, testicular tissue from abnormal spermatogenesis was analysed. Methods Testicular treatment biopsies were obtained from 27 men. Five presented oligozoospermia, 9 obstructive azoospermia (4 congenital bilateral absence of the vas deferens; 5 secondary azoospermia) and 13 nonobstructive azoospermia (5 hypospermatogenis; 3 maturation arrest; 5 Sertoli-cell-only syndrome). Immunohistochemical staining was performed for active caspases-3, −8 and −9. The presence of active caspases in Sertoli cells and germ cells was analyzed using stereological tools. Results Increased active caspase-3 was found in Sertoli-cellonly syndrome. No significant differences were found in maturation arrest. In hypospermatogenesis, primary spermatocytes were the germ cells with higher active caspases. Oligozoospermia and secondary obstruction showed significant differences among germ cells for the presence of all active caspases. In oligozoospermia, spermatogonia presented significant increased active caspase-9 in relation to active caspase-8. In primary obstruction and hypospermatogenesis, germ cells presented significant increased active caspases-3 and −9. Conclusions Results suggest that increased active caspase-3 might be involved in Sertoli-cell-only syndrome etiology. In cases of hypospermatogenesis, intrinsic lesions at the meiotic stage seem to be related to the pathology. In secondary obstruction apoptosis is suggested to be initiated due to extrinsic and intrinsic lesions, whereas in primary obstruction only the intrinsic apoptotic pathway seems to be present. Finally, in oligozoospermic patients spermatogonia death by mitochondrial damage additionally to meiosis malfunctioning, might be on the origin of the decreased sperm output.

“…Normally, breaks are physiologically reassembled before spermatozoa are released from germinal epithelium [27], but, as a consequence of testicular injuries, including heat, exposure to toxicant [26,31,34] radiation [37], withdrawal of hormonal support [22,55,61], other testicular pathologies [18] or life habits (smoke, drug use), the repair process could be impaired, so that spermatozoa show high levels of DNA damage [8,49].…”

Section: Discussionmentioning

confidence: 99%

Lower sperm DNA fragmentation after r-FSH administration in functional hypogonadotropic hypogonadism

Ruvolo

Roccheri

Brucculeri

et al. 2013

Purpose An observational clinical and molecular study was designed to evaluate the effects of the administration of recombinant human FSH on sperm DNA fragmentation in men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. Methods In the study were included 53 men with a nonclassical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. In all patients, sperm DNA fragmentation index (DFI), assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) in situ DNA nick end-labelling (TUNEL) assay, was evaluated before starting the treatment with 150 IU of recombinant human FSH, given three times a week for at least 3 months. Patients' semen analysis and DNA fragmentation index were re-evaluated after the 3-month treatment period. Results After recombinant human FSH therapy, we did not find any differences in terms of sperm count, motility and morphology. The average DNA fragmentation index was significantly reduced (21.15 vs 15.2, p<0.05), but we found a significant reduction in patients with high basal DFI values (>15 %), while no significant variation occurred in the patients with DFI values ≤15 %. Conclusions Recombinant human FSH administration improves sperm DNA integrity in hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia men with DNA fragmentation index value >15 % .