FE-SEM Characterization of <i>α</i>-Mannose Density and Surface Mapping Changes in Human Sperm Head During <i>In Vitro</i> Capacitation

Gómez‐Torres, María José; Robles-Gómez, Laura; Huerta-Retamal, Natalia; Sáez‐Espinosa, Paula; Sánchez, Manuel A. Bueno; Aizpurúa, Jon; Romero, Alejandro

doi:10.1017/s1431927620024630

Cited by 5 publications

(6 citation statements)

References 32 publications

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“…These outcomes suggest the differentiated response of the glycocalyx components to capacitation time. In this context, the authors of this current paper previously described a decrease in D-mannose residues during capacitation using the paired FE-SEM methodology [ 14 ]. Specifically, the density of D-mannose decreased significantly between the in vitro capacitated sperm during 1 (CAP1) and 4 h (CAP4), showing a clear time-dependent change.…”

Section: Discussionmentioning

confidence: 99%

“…The AAA lectin was used due to the preference for fucose and terminal fucose residues on complex oligosaccharides and glycoconjugates [ 27 , 28 ]. The lectin labelling protocol was adapted from [ 14 ]. A total volume of 5 µL from each glutaraldehyde-fixed condition was placed on a round coverslip with a diameter of 12 mm.…”

Section: Methodsmentioning

confidence: 99%

“…A total of 105 spermatozoa head digital micrographs (1024 × 768 pixels) were obtained at 18,000× magnification for each physiological condition. The calibration and recording of the density and topographical distribution of nanoparticles were previously described [ 14 ]. Briefly, calibrated micrographs were edited using Adobe Photoshop ® CS5 (Adobe Inc., San Jose, CA, USA) to obtain oriented images and the SigmaScan ® Pro (SPSSTM, Chicago, IL, USA) digital imaging software to count gold nanoparticles and to consider their topographic spatial distribution in both acrosomal and postacrosomal regions.…”

Section: Methodsmentioning

confidence: 99%

“…In this context, the authors of this paper previously used field emission scanning electron microscopy (FE-SEM) to accurately localize and quantify mannose residues in human sperm surface during in vitro capacitation. Moreover, a significant and time-dependent decrease of mannose was registered during in vitro capacitation and a relocation mostly affecting acrosomal domain apical areas [ 14 ]. These findings showed the efficacy of FE-SEM and gold labelling as a useful tool for studying sperm surface components.…”

Section: Introductionmentioning

confidence: 99%

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Quantification and Topographical Distribution of Terminal and Linked Fucose Residues in Human Spermatozoa by Using Field Emission Scanning Electron Microscopy (FE-SEM)

Robles-Gómez

Sáez‐Espinosa

López-Viloria

et al. 2021

IJMS

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The modification of sperm glycocalyx is an essential process during sperm capacitation. The presence and redistribution of terminal and linked fucose have been described during in vitro capacitation in humans. However, the influence of the capacitation time on the quantification and localization of terminal and linked fucose is still unknown. In this study, the quantitative and qualitative changes in fucosyl residues during different in vitro capacitation times (1 and 4 h), are simultaneously characterized by using Aleuria aurantia (AAA) lectin–gold labelling and high-resolution field emission scanning electron microscopy (FE-SEM) in human sperm. A significant decrease was found in the number of terminal fucose registered in the whole sperm head during the in vitro capacitation. Nevertheless, the quantification of fucose residues after 1 h of in vitro capacitation was very similar to those found after 4 h. Therefore, the changes observed in terminal and linked fucose during capacitation were not time-dependent. Furthermore, the comprehensive analysis of the topographic distribution showed the preferential fucosyl location in the acrosomal region and the presence of distinct clusters distributed over the head in all the studied conditions. Overall, these findings corroborate the validity of FE-SEM combined with gold labelling to register changes in surface molecules during in vitro sperm capacitation.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Quantification and Topographical Distribution of Terminal and Linked Fucose Residues in Human Spermatozoa by Using Field Emission Scanning Electron Microscopy (FE-SEM)

Robles-Gómez

Sáez‐Espinosa

López-Viloria

et al. 2021

IJMS

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“…A comprehensive study of morphological features was performed using FE-SEM. For FE-SEM analysis, a total of 5 µL of glutaraldehyde-fixed sperm suspension was placed on a round coverslip with a diameter of 12 mm and air-dried [56]. At that time, the coverslip was rehydrated three times with PBS for 5 min.…”

Section: Morphological and Ultrastructural Characterizationmentioning

confidence: 99%

Immunofluorescence and High-Resolution Microscopy Reveal New Insights in Human Globozoospermia

Sáez‐Espinosa

Robles-Gómez

Ortega-López

et al. 2022

IJMS

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Globozoospermia is a rare and severe type of teratozoospermia characterized by the presence of round-headed, acrosomeless spermatozoa with cytoskeleton defects. Current data support a negative relationship between globozoospermia and intracytoplasmic sperm injection (ICSI) outcomes, revealing the need to perform exhaustive studies on this type of sperm disorder. The aim of this study was to evaluate different structural, functional and molecular sperm biomarkers in total globozoospermia with proper embryo development after ICSI. The combination of field-emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) allowed us to identify and correlate eight morphological patterns with both types of microscopy. Additionally, results reported a high percentage of coiled forms, with cytoplasmic retentions around the head and midpiece. By fluorescent microscopy, we detected that most of the sperm showed tubulin in the terminal piece of the flagellum and less than 1% displayed tyrosine phosphorylation in the flagellum. Moreover, we did not detect chaperone Heat shock-related 70 kDa protein 2 (HSPA2) in 85% of the cells. Overall, these findings provide new insights into globozoospermia, which could have potential implications in improving sperm selection methods for assisted reproductive techniques.

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A comprehensive N-glycome map of porcine sperm membrane before and after capacitation

Li,

Zhai,

et al. 2024

Carbohydrate Polymers

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FE-SEM Characterization of α-Mannose Density and Surface Mapping Changes in Human Sperm Head During In Vitro Capacitation

Abstract: Abstract

Cited by 5 publications

References 32 publications

Quantification and Topographical Distribution of Terminal and Linked Fucose Residues in Human Spermatozoa by Using Field Emission Scanning Electron Microscopy (FE-SEM)

Quantification and Topographical Distribution of Terminal and Linked Fucose Residues in Human Spermatozoa by Using Field Emission Scanning Electron Microscopy (FE-SEM)

Immunofluorescence and High-Resolution Microscopy Reveal New Insights in Human Globozoospermia

A comprehensive N-glycome map of porcine sperm membrane before and after capacitation

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