Abstract:SummaryThe principal aim of this retrospective study was to examine the relationship between sperm apoptotic biomarkers and the patient's biclinical characteristics, the conven-
“…To detect apoptosis in ejaculated spermatozoa, we used Annexin V‐Cy3 in conjunction with a vital dye (CFDA), allowing to detect early and late apoptosis (or necrosis) 12,46,47 . The percentages of both necrotic and early apoptotic spermatozoa were significantly associated with the rate of atypical sperm forms and the rate of various forms of sperm abnormalities including head and tail.…”
Section: Discussionmentioning
confidence: 99%
“…To be treated as morphologically normal, a spermatozoon would have a normal acrosome, an oval head between 2.5 and 3.5 μm width and 5 and 6 μm long, a midpiece of 4.0 to 5.0 μm, and a tail about 50 μm long 8 . Several reports have found higher rates of DNA damage, sperm aneuploidy, and protamination defects in infertile men with impaired semen parameters in comparison with controls and their association with morphological abnormalities 2,9‐12 …”
Section: Introductionmentioning
confidence: 99%
“…On the other hand, many data indicate that poor sperm morphology is associated with higher rates of caspases’ expression, 24,25 suggesting an association between teratozoospermia and apoptosis. Many reports have indicated the occurrence of apoptosis‐like characteristics, including DNA breakage, presence of activated caspases, and externalization of the phosphatidylserine (PS) on the sperm plasma membrane in ejaculated human spermatozoa of infertile men with different potential of infertility, 12,26‐28 however, the link between apoptosis and teratozoospermia has been poorly investigated so far.…”
This study was carried out to evaluate the level of nuclear sperm DNA damage in men with isolated polymorphic teratozoospermia and examining its relationship with apoptosis and oxidative stress. A total of 89 subjects were divided into two groups: men with isolated teratozoospermia (n = 69) and men with normal semen parameters (n = 20) as controls. Sperm DNA breaks were determined by using acridine orange staining. The proportion of viable spermatozoa with mitochondrial transmembrane depolarization was detected by fluorescence microscopy through the use of MitoPTJC‐1 staining method. Bivariate Annexin V/6‐CFDA analysis was then set out in order to measure the percentage of both viable and dead spermatozoa with phosphatidylserine (PS) externalization. Seminal antioxidant profile (reduced glutathione (GSHr); oxidized glutathione (GSSG); glutathione S‐transferase (GST)) and total protein sulfhydryl (P‐SH) concentrations were measured spectrophotometrically. Men with isolated teratozoospermia, when compared to controls, showed significantly increased level of single sperm DNA breaks and higher proportions of spermatozoa with phosphatidylserine externalization and mitochondrial depolarization. Among the differently studied oxidative stress seminal parameters, the rates of seminal GSHr, GST, and P‐SH were significantly decreased in the teratozoospermic group. However, the seminal rates of GSSG and GST have decreased, but only GST did not show a significant difference. Interestingly, significant correlations were found between the studied apoptotic markers and the rate of atypical sperm forms with the incidences of head abnormalities. Furthermore, positive inter‐correlations were found between sperm DNA defects, impaired seminal antioxidant status, and the apoptotic sperm markers. Such data provide clear evidence that the apoptotic alterations are closely correlated to abnormal sperm morphology and DNA damage. Moreover, decreased seminal antioxidant profile may be a crucial factor involved in the mechanism of sperm cell death‐mediated DNA breaks in teratozoospermic semen.
“…To detect apoptosis in ejaculated spermatozoa, we used Annexin V‐Cy3 in conjunction with a vital dye (CFDA), allowing to detect early and late apoptosis (or necrosis) 12,46,47 . The percentages of both necrotic and early apoptotic spermatozoa were significantly associated with the rate of atypical sperm forms and the rate of various forms of sperm abnormalities including head and tail.…”
Section: Discussionmentioning
confidence: 99%
“…To be treated as morphologically normal, a spermatozoon would have a normal acrosome, an oval head between 2.5 and 3.5 μm width and 5 and 6 μm long, a midpiece of 4.0 to 5.0 μm, and a tail about 50 μm long 8 . Several reports have found higher rates of DNA damage, sperm aneuploidy, and protamination defects in infertile men with impaired semen parameters in comparison with controls and their association with morphological abnormalities 2,9‐12 …”
Section: Introductionmentioning
confidence: 99%
“…On the other hand, many data indicate that poor sperm morphology is associated with higher rates of caspases’ expression, 24,25 suggesting an association between teratozoospermia and apoptosis. Many reports have indicated the occurrence of apoptosis‐like characteristics, including DNA breakage, presence of activated caspases, and externalization of the phosphatidylserine (PS) on the sperm plasma membrane in ejaculated human spermatozoa of infertile men with different potential of infertility, 12,26‐28 however, the link between apoptosis and teratozoospermia has been poorly investigated so far.…”
This study was carried out to evaluate the level of nuclear sperm DNA damage in men with isolated polymorphic teratozoospermia and examining its relationship with apoptosis and oxidative stress. A total of 89 subjects were divided into two groups: men with isolated teratozoospermia (n = 69) and men with normal semen parameters (n = 20) as controls. Sperm DNA breaks were determined by using acridine orange staining. The proportion of viable spermatozoa with mitochondrial transmembrane depolarization was detected by fluorescence microscopy through the use of MitoPTJC‐1 staining method. Bivariate Annexin V/6‐CFDA analysis was then set out in order to measure the percentage of both viable and dead spermatozoa with phosphatidylserine (PS) externalization. Seminal antioxidant profile (reduced glutathione (GSHr); oxidized glutathione (GSSG); glutathione S‐transferase (GST)) and total protein sulfhydryl (P‐SH) concentrations were measured spectrophotometrically. Men with isolated teratozoospermia, when compared to controls, showed significantly increased level of single sperm DNA breaks and higher proportions of spermatozoa with phosphatidylserine externalization and mitochondrial depolarization. Among the differently studied oxidative stress seminal parameters, the rates of seminal GSHr, GST, and P‐SH were significantly decreased in the teratozoospermic group. However, the seminal rates of GSSG and GST have decreased, but only GST did not show a significant difference. Interestingly, significant correlations were found between the studied apoptotic markers and the rate of atypical sperm forms with the incidences of head abnormalities. Furthermore, positive inter‐correlations were found between sperm DNA defects, impaired seminal antioxidant status, and the apoptotic sperm markers. Such data provide clear evidence that the apoptotic alterations are closely correlated to abnormal sperm morphology and DNA damage. Moreover, decreased seminal antioxidant profile may be a crucial factor involved in the mechanism of sperm cell death‐mediated DNA breaks in teratozoospermic semen.
“…1c, d). Although PtdSer has been noted on sperm previously 12–14 , it was considered to mark dead or non-viable sperm because PtdSer is a key eat-me signal on cells undergoing apoptosis, which facilitates recognition and uptake by phagocytes 15 . Interestingly, PtdSer can also be transiently exposed on viable cells in certain conditions (such as activated B and T cells, myoblasts, or macrophages 16–19 ), and therefore, we further investigated the relevance of PtdSer exposure on the sperm.Fig.…”
Fertilization is essential for species survival. Although Izumo1 and Juno are critical for initial interaction between gametes, additional molecules necessary for sperm:egg fusion on both the sperm and the oocyte remain to be defined. Here, we show that phosphatidylserine (PtdSer) is exposed on the head region of viable and motile sperm, with PtdSer exposure progressively increasing during sperm transit through the epididymis. Functionally, masking phosphatidylserine on sperm via three different approaches inhibits fertilization. On the oocyte, phosphatidylserine recognition receptors BAI1, CD36, Tim-4, and Mer-TK contribute to fertilization. Further, oocytes lacking the cytoplasmic ELMO1, or functional disruption of RAC1 (both of which signal downstream of BAI1/BAI3), also affect sperm entry into oocytes. Intriguingly, mammalian sperm could fuse with skeletal myoblasts, requiring PtdSer on sperm and BAI1/3, ELMO2, RAC1 in myoblasts. Collectively, these data identify phosphatidylserine on viable sperm and PtdSer recognition receptors on oocytes as key players in sperm:egg fusion.
“…Among the physiological properties involved in sperm function, apoptosis has been proposed as one of the more detrimental to spermatozoa’s fertilization potential. An increased presence of apoptotic markers activated caspase-3, externalized phosphatidylserine, or fragmented spermatic DNA has been linked to abnormal sperm motility or morphology [ 3 , 4 , 5 , 6 , 7 , 8 ], a decrease in fertilization rate and optimal quality embryos in couples with normozoospermic men [ 9 ], a decrease in fertilization potential, and a reduced ability to trigger acrosome reaction [ 10 ]. Therefore, the separation of sperm with initiated apoptosis is interesting to ensure the selection of the most physiologically competent sperm.…”
The application of MACS non-apoptotic sperm selection in infertility clinics is controversial since the published literature does not agree on its effect on reproductive outcomes. Therefore, it is not part of the routine clinical practice. Classical measures of reproductive success (pregnancy or live birth rates per ovarian stimulation) introduce a bias in the evaluation of a technique’s effect, since only the best embryo is transferred. This retrospective, multicenter, observational study evaluated the impact of MACS on reproductive outcomes, measuring results in classical parameters and cumulative live birth rates (CLBR). Data from ICSI cycles using autologous oocyte in Spanish IVIRMA fertility clinics from January 2008 to February 2020 were divided into two groups according to their semen processing: standard practice (reference: 46,807 patients) versus an added MACS sperm selection (1779 patients). Only when measured as CLBR per embryo transferred and per MII oocyte used was the difference between groups statistically significant. There were no significant differences between MACS and reference groups on pregnancy and live birth rates. In conclusion, results suggest that non-apoptotic sperm selection by MACS on unselected males prior to ICSI with autologous oocytes has limited clinical impact, showing a subtle increase in CLBR per embryo transferred.
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