Laser-assisted selection of immotile spermatozoa has no effect on obstetric and neonatal outcomes of TESA-ICSI pregnancies

Background Azoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable, immotile spermatozoa before the procedure. There are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI. Methods A retrospective comparative study was conducted on outcomes of ICSI cycles with testicular spermatozoa from June 2014 to June 2018. Of 132 cycles, 33 were allocated to the test group and oocytes were injected with immotile spermatozoa selected by laser, 99 cycles were allocated as control group. Results Compared with the control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the test group in either fresh or frozen transfer cycles. The cumulative live birth rate in the test group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight, and the malformation rate between the test and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05). Conclusions No negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.

Successful birth after ICSI with testicular immotile spermatozoa from a patient with total MMAF in the ejaculates: a case report

Summary There has been no report on the outcome of vitrified blastocyst transfer from a vitrified oocyte injected with immotile testicular spermatozoa with only multiple morphological abnormalities of the sperm flagella (MMAF). A couple diagnosed with MMAF returned to the clinic to attempt pregnancy using their vitrified oocytes. Testicular spermatozoa were injected intracytoplasmically, and the following intracytoplasmic sperm injection results were observed. In the second cycle, surplus vitrified oocytes and testicular retrieved sperm were used, but no pregnancy ensued. In the third cycle, a surplus vitrified blastocyst was transferred, and a healthy female child was delivered, with a birth weight of 3050 g and a birth length of 53 cm. In this report we describe a successful pregnancy achieved in a patient presenting MMAF. The successful pregnancy was obtained from vitrified oocytes microinjected with testicular retrieved sperm in a vitrified blastocyst transfer.

Laser-assisted selection of immotile spermatozoa has no effect on obstetric and neonatal outcomes of TESA-ICSI pregnancies

Background Azoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable but immotile spermatozoa before the procedure. However, there are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI. Methods A retrospective comparative study was conducted on patients who underwent ICSI treatment with testicular spermatozoa in our Reproductive Medicine Unit from June 2014 to June 2018. The 132 cycles were divided into two groups according to whether laser-assisted selection of spermatozoa was used. Results Compared with control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the laser group in either fresh or frozen transfer cycles. The cumulative live birth rate in the laser group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight and the malformation rate between the laser and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05). Conclusions No negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.

ATP activation of peritubular cells drives testicular sperm transport

Spermatogenesis, the complex process of male germ cell proliferation, differentiation, and maturation, is the basis of male fertility. In the seminiferous tubules of the testes, spermatozoa are constantly generated from spermatogonial stem cells through a stereotyped sequence of mitotic and meiotic divisions. The basic physiological principles, however, that control both maturation and luminal transport of the still immotile spermatozoa within the seminiferous tubules remain poorly, if at all, defined. Here, we show that coordinated contractions of smooth muscle-like testicular peritubular cells provide the propulsive force for luminal sperm transport toward the rete testis. Using a mouse model for in vivo imaging, we describe and quantify spontaneous tubular contractions and show a causal relationship between peritubular Ca2+ waves and peristaltic transport. Moreover, we identify P2 receptor-dependent purinergic signaling pathways as physiological triggers of tubular contractions both in vitro and in vivo. When challenged with extracellular ATP, transport of luminal content inside the seminiferous tubules displays stage-dependent directionality. We thus suggest that paracrine purinergic signaling coordinates peristaltic recurrent contractions of the mouse seminiferous tubules to propel immotile spermatozoa to the rete testis.

ATP activation of peritubular cells drives testicular sperm transport

Spermatogenesis, the complex developmental process of male germ cell proliferation, differentiation, and maturation, is the basis of male fertility and reproductive fitness. In the seminiferous tubules of the testes, spermatozoa are constantly generated from spermatogonial stem cells through a stereotyped sequence of mitotic and meiotic divisions. The basic physiological principles, however, that control both maturation and luminal transport of the still immotile spermatozoa within the seminiferous tubules remain poorly, if at all, defined. Here, we show that coordinated contractions of smooth muscle-like testicular peritubular cells provide the propulsive force for luminal sperm transport towards the rete testis and epididymis. Using a mouse model for in vivo imaging, we describe and quantify spontaneous tubular contractions and show a causal relationship between peritubular Ca2+ waves and peristaltic transport. Moreover, we identify P2 receptor-dependent purinergic signaling pathways as physiological triggers of tubular contractions both in vitro and in vivo. When challenged with extracellular ATP, transport of luminal content inside the seminiferous tubules displays stage-dependent directionality. We thus suggest that paracrine purinergic signaling coordinates peristaltic recurrent contractions of the mouse seminiferous tubules to propel immotile spermatozoa to the rete testis. Consequently, our findings could have substantial pharmaceutical implications for both infertility treatment and / or male contraception.

The SEPT12 complex is required for the establishment of a functional sperm head–tail junction

The connecting pieces of the sperm neck link the flagellum and the sperm head, and they are important for initiating flagellar beating. The connecting pieces are important building blocks for the sperm neck; however, the mechanism of connecting piece assembly is poorly understood. In the present study, we explored the role of septins in sperm motility and found that Sept12D197N knock-in (KI) mice produce acephalic and immotile spermatozoa. Electron microscopy analysis showed defective connecting pieces in sperm from KI mice, indicating that SEPT12 is required for the establishment of connecting pieces. We also found that SEPT12 formed a complex with SEPT1, SEPT2, SEPT10 and SEPT11 at the sperm neck and that the D197N mutation disrupted the complex, suggesting that the SEPT12 complex is involved in the assembly of connecting pieces. Additionally, we found that SEPT12 interacted and colocalized with γ-tubulin in elongating spermatids, implying that SEPT12 and pericentriolar materials jointly contribute to the formation of connecting pieces. Collectively, our findings suggest that SEPT12 is required for the formation of striated columns, and the capitulum and for maintaining the stability of the sperm head–tail junction.

Frozen semen quality and fertility of imported pure Holstein Friesian and Sahiwal breeding bulls in Bangladesh

The present study was conducted to evaluate the frozen semen quality in terms of different motility patterns and fertility of imported pure Holstein Friesian and Sahiwal breeding bulls maintained under farming condition of Bangladesh. 100% pure Holstein Friesian and Sahiwal breeding bulls were imported and thereafter reared in the American dairy limited (ADL). Semen samples were collected once a week, processed and frozen in the laboratory of ADL. On the day of collection, semen volume and sperm concentration measured immediately after collection. Different motility patterns of spermatozoa (progressive, fast, slow, local, immotile) were measured by Computer Assisted Semen Analyzer (CASA). Effect of breed was found significant (p<0.05) on semen volume and sperm concentration. The semen volume (ml/ejaculate) and sperm concentration (×106) were found to be 6.13±0.28 and 1664.28±62.41 in Holstein Friesian breeding bulls whereas 5.26±0.17 and 2036.00±43.99 in Sahiwal bulls, respectively. The progressive motility, fast motility, slow motility and local motility were significantly (p<0.05) higher in Holstein Friesian bulls than that of Sahiwal bulls both before and after freezing. Different motility patterns such as progressive, fast, slow, local and immotile motility were found to be 89.22±0.45%, 75.97±0.67%, 9.10±0.36%, 4.25±0.15% and 10.05±0.41% in Holstein Friesian bulls whereas 84.98±0.89%, 65.99±0.92%, 11.88±0.37%, 5.65±0.19% and 15.69±0.54% in Sahiwal breeding bulls, respectively before freezing. On the other hand, after freezing, the progressive, fast, slow, local and immotile spermatozoa were found to be 68.19±0.46%, 53.39±2.01%, 9.21±0.46%, 8.47±0.72% and 26.76±2.01% in Holstein Friesian bulls whereas 56.54±0.25%, 42.52±0.67%, 11.11±0.55%, 13.04±1.24% and 30.45±1.58% in Sahiwal bulls, respectively. Non-return rate was found insignificant between the breeds and it was 64.19% in Holstein Friesian and 64.71% in Sahiwal bulls. In conclusion, frozen semen quality of Holstein Friesian pure breed is better than that of Sahiwal breeding bulls but fertility of both breeds are similar. Therefore, frozen semen of both breeds can be used in the field level for the genetic improvement of cattle. Asian Australas. J. Biosci. Biotechnol. 2020, 5 (1), 33-41

Human Sperm Motility, Viability, and Morphology Decreased after Cryopreservation

The aim of this study was to analyze human sperm motility, viability, and morphology before and after cryopreservation. This true laboratory experimental study had pre and post randomized one group design. The study was conducted at the Embryology, Andrology, and Genetics Laboratory, Department of Medical Biology, Faculty of Medicine, Universitas Airlangga from August to November 2017. The eighteen samples of fresh semen were collected from male volunteers who agreed and signed the informed consent of the study. Samples were analyzed their motility, viability, and morphology before and after cryopreservation. Results of this study indicated differentiation between motility before and after cryopreservation. Cryopreservation process decreased progressive motility (42.22 + 9.46%; 17.83 + 6.24%; p< 0.0001) and increased the number of immotile spermatozoa (35.44 + 10.15%; 60.11 + 12.53%; p< 0.0001). Cryopreservation also decreased human sperm viability (73.78 + 8.91%; 40.83 + 12.89%; p< 0.0001) and morphology (10.94 + 4.96%; 7.39 + 3.90%; p< 0.0001). Cryopreservation of human spermatozoa caused the decreased of motility, viability, and morphology.