Effects on glycocalyx structures of frozen-thawed bovine sperm induced by flow cytometry and artificial capacitation

Umezu, Kohei; Hiradate, Yuuki; Numabe, Takashi; Hara, Kenshiro; Tanemura, Kentaro

doi:10.1262/jrd.2017-065

Cited by 12 publications

(12 citation statements)

References 43 publications

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“…It appears to be the case because these patterns were differentially present in the spermatozoa from HF and LF bulls, although the differences were non-significant ( Supplementary Results: Figure 10 ). Multiple patterns of lectin staining have earlier been reported in the spermatozoa of many species including bovine ( 74 ), bats ( 13 ), human and mouse ( 56 , 75 , 89 , 90 ) providing evidence for the existence of sub-populations in the ejaculated semen. Moreover, the pattern of lectin binding observed in this study differs from what has been observed in cattle and humans ( 3 , 74 , 75 ) highlighting species-specific glycan distribution on sperm-surface.…”

Section: Discussionmentioning

confidence: 88%

“…At the time of capacitation, the sperm surface undergoes extensive remodeling/reorganization especially with respect to the modifications and alterations of the GCs ( 74 ). To detect the changes in abundance of sperm glycans after induction of in vitro capacitation, the fluorescence signal produced after lectin/s binding were quantified, for the non-capacitated (in NCM) and capacitated (in CM) buffalo bull spermatozoa, in the same manner as already explained.…”

Section: Resultsmentioning

confidence: 99%

“…The dramatic biochemical and extensive modifications including the re-organization and alteration of the GCs that occurs during capacitation prepare the sperm for successful fertilization ( 7 , 36 , 74 , 95 ). In the present study, a significant reduction in the MFI upon JAC and ABL, MAL II and LEL binding was observed in the in vitro capacitated spermatozoa vis-à-vis the non-capacitated spermatozoa of the HF buffalo bulls.…”

Section: Discussionmentioning

confidence: 99%

“…These results underpinned the proposition of progressive removal/re-organization of their cognate receptor ligands of the lectins (glycans) from the sperm plasma membrane after capacitation. The changes in the structures of the GCs i.e., the changes in the molecular arrangements of the glycocalyx (SpAGT) associated with perturbations such as capacitation ( 11 , 74 ), cryopreservation ( 8 , 34 ), flow cytometry ( 35 , 74 ), or removal of glycans by using enzymes ( 8 , 53 ) have successfully been monitored and well documented using lectins as reporters. Overall, our data reveal that both the O-linked and N-linked glycans were either removed or extensively re-organized during the in vitro induced capacitation of the buffalo spermatozoa, thus possibly affecting the overall topography of its glycocalyx.…”

Section: Discussionmentioning

confidence: 99%

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A Higher Abundance of O-Linked Glycans Confers a Selective Advantage to High Fertile Buffalo Spermatozoa for Immune-Evasion From Neutrophils

et al. 2020

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The glycans on the plasma membrane of cells manifest as the glycocalyx, which serves as an information-rich frontier that is directly in contact with its immediate milieu. The glycoconjugates (GCs) that adorn most of the mammalian cells are also abundant in gametes, especially the spermatozoa where they perform unique reproduction-specific functions e.g., inter-cellular recognition and communication. This study aimed to implicate the sperm glycosylation pattern as one of the factors responsible for low conception rates observed in buffalo bulls. We hypothesized that a differential abundance of glycans exists on the spermatozoa from bulls of contrasting fertilizing abilities endowing them with differential immune evasion abilities. Therefore, we investigated the role of glycan abundance in the phagocytosis and NETosis rates exhibited by female neutrophils (PMNs) upon exposure to such spermatozoa. Our results indicated that the spermatozoa from high fertile (HF) bulls possessed a higher abundance of O-linked glycans e.g., galactosyl (β-1,3)N-acetylgalactosamine and N-linked glycans like [GlcNAc]1-3, N-acetylglucosamine than the low fertile (LF) bull spermatozoa. This differential glycomic endowment appeared to affect the spermiophagy and NETosis rates exhibited by the female neutrophil cells (PMNs). The mean percentage of phagocytizing PMNs was significantly different ( P < 0.0001) for HF and LF bulls, 28.44 and 59.59%, respectively. Furthermore, any introduced perturbations in the inherent sperm glycan arrangements promoted phagocytosis by PMNs. For example, after in vitro capacitation the mean phagocytosis rate (MPR) rate in spermatozoa from HF bulls significantly increased to 66.49% ( P < 0.01). Likewise, the MPR increased to 70.63% ( p < 0.01) after O-glycosidase & α2-3,6,8,9 Neuraminidase A treatment of spermatozoa from HF bulls. Moreover, the percentage of PMNs forming neutrophil extracellular traps (NETs) was significantly higher, 41.47% when exposed to spermatozoa from LF bulls vis-à-vis the spermatozoa from HF bulls, 15.46% ( P < 0.0001). This is a pioneer report specifically demonstrating the role of O-linked glycans in the immune responses mounted against spermatozoa. Nevertheless, further studies are warranted to provide the measures to diagnose the sub-fertile phenotype thus preventing the losses incurred by incorrect selection of morphologically normal sperm in the AI/IVF reproduction techniques.

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Section: Discussionmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

A Higher Abundance of O-Linked Glycans Confers a Selective Advantage to High Fertile Buffalo Spermatozoa for Immune-Evasion From Neutrophils

et al. 2020

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“…Later on, the applications of flow cytometry expanded to the study of the integrity of the membrane, acrosome and mitochondrial function (Engh, Clausen, & Purvis, ; Evenson, Darzynkiewicz, & Melamed, ; Graham, Kunze, & Hammerstedt, ; Miyazaki, Fukuda, Takeuchi, Itoh, & Takada, ; Ronot & Auger, ). The possibility to detect differences in DNA content allowed the development of the sexed semen market in the bovine industry (Johnson, Flook, & Look, ; Morrell, Keeler, Noakes, Mackenzie, & Dresser, ; Sharpe & Evans, ; Tubman, Brink, Suh, & Seidel, ; Umezu, Hiradate, Numabe, Hara, & Tanemura, ; Vazquez et al., ). While the costs and complexity of initial flow cytometers limited their use to specialized laboratories and mainly to research applications, nowadays more affordable, reliable and user friendly systems allow to implement flow cytometry in the clinical setting in many andrology laboratories.…”

Section: Introductionmentioning

confidence: 99%

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Peña

Rodriguez

Gil

et al. 2018

Reprod Domestic Animals

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Contents Flow cytometry is increasingly used in research and also in clinical andrology. Recent developments in instrumentation, availability of probes and bioinformatics expand the possibilities of flow cytometry well beyond the classical two parametric analyses in use. In this paper, an overview of recent developments in flow cytometry will be presented under the perspective of the authors; aspects such a multicolor assays and computational cytometry will be discussed as well.

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Developing glycoproteomics reveals the role of posttranslational glycosylation in the physiological and pathological processes of male reproduction

Cheng,

Luo,

et al. 2024

iMetaOmics

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Glycosylation plays a pivotal role in the physiological and pathological processes of male reproduction. It impacts thousands of proteins and is actively ongoing through all stages of reproduction, including spermatogenesis, maturation, capacitation, and fertilization. However, our grasp on glycosylation within male reproductive processes remains limited, largely due to the technical hurdles. Recent advancements have seen the mapping of the glycoproteome of human semen, utilizing cutting‐edge glycoproteomic technologies. This breakthrough lays the groundwork for in‐depth research into the influence of glycosylation on male reproductive system and related disorders. Nevertheless, the field faces numerous challenges that necessitate further advancements in glycoproteomic methodologies. In this analysis, we evaluated the potential applications of advanced glycoproteomic techniques in the study of male reproduction and summarized the detailed profiling of the human semen glycome and glycoproteome. Our current understanding of glycosylation's role within the male reproductive system alongside recent progress in glycoproteomics may equip biologists with a comprehensive insight. Furthermore, this analysis brought together findings on abnormal glycosylation and its link to male reproductive disorders in the view of glycomics and glycoproteomics. It can facilitate the clinical application of glyco‐related biomarkers and targets in the treatment of infertility.

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Effects on glycocalyx structures of frozen-thawed bovine sperm induced by flow cytometry and artificial capacitation

Cited by 12 publications

References 43 publications

A Higher Abundance of O-Linked Glycans Confers a Selective Advantage to High Fertile Buffalo Spermatozoa for Immune-Evasion From Neutrophils

A Higher Abundance of O-Linked Glycans Confers a Selective Advantage to High Fertile Buffalo Spermatozoa for Immune-Evasion From Neutrophils

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Developing glycoproteomics reveals the role of posttranslational glycosylation in the physiological and pathological processes of male reproduction

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