Separation of acrosome-reacted viable sperm is an important problem in the analysis of sperm function and the micromanipulation of gametes such as the microinjection of sperm into ooplasm and subzonal sperm insertion. In the present study, we collected acrosome-reacted sperm selectively using paramagnetic immunobeads, referred to as MH61-beads, which attach themselves to the head of the acrosome-reacted sperm and are recovered by a magnet. The maximum number of sperm was recovered by the MH61-beads when 3 x 10(5) sperm/100 microliters and 6 x 10(5) MH61-beads were coincubated for 1 hour. The recovery of sperm was significantly correlated with the percentage of the acrosome reaction in the capacitation medium. After incubation for 12 hours, 2.4-9.0 x 10(4) sperm could be collected, and the percentage of viable and acrosome-reacted sperm was 28-60%. To improve the recovery of the acrosome-reacted sperm, we utilized follicular fluid, which induces the acrosome reaction. Addition of follicular fluid to the capacitation medium at a final concentration of 20% induced the acrosome reaction and increased the recovery of acrosome-reacted sperm significantly. In conclusion, separation of acrosome-reacted sperm by the MH61-bead-binding method could become a useful technique in research of sperm function and gamete micromanipulation.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Role of Rab proteins in PFOA-induced changes in boar sperm motility and capacitation.
Reprod Toxicol
December 2024
Department of Animal Science and Biotechnology, Kyungpook National University, Sangju, Gyeongsangbuk-do 37224, Republic of Korea; Research Institute for Innovative Animal Science, Kyungpook National University, Sangju, Gyeongsangbuk-do 37224, Republic of Korea. Electronic address:
Article Synopsis
- - The study investigates how perfluorooctanoic acid (PFOA), an environmental contaminant, negatively impacts sperm functions like motility and capacitation in boar sperm by examining specific proteins called Rab proteins.
- - Findings show that exposure to PFOA significantly reduces sperm motility and is linked to changes in Rab proteins, particularly Rab14, Rab25, and Rab34, which are crucial for sperm development and the acrosome reaction.
- - The research suggests that the alterations in Rab protein activity could serve as biomarkers for measuring the reproductive toxicity of environmental toxins like PFOA, potentially leading to lower pregnancy rates.
Lectin Functionalised Iron Magnetic Nanoparticle-Based Sperm Selection: A Potential Technique to Improve Bull Sperm Quality In Vitro.
Reprod Domest Anim
October 2024
Theriogenology Laboratory, Southern Regional Station of ICAR-National Dairy Research Institute, Bengaluru, Karnataka, India.
Premature acrosomal exocytosis in cryopreserved semen is one of the reasons attributed to low fertility among livestock. In the present study, we attempted to enhance the cryopreserved semen quality by selective removal of acrosome-reacted spermatozoa using FITC-PNA conjugated iron magnetic nanoparticles (MNPs). Further, the effect of nano purification on other sperm functional attributes was also assessed.
View Article and Find Full Text PDFIn vitro versus cryo-induced capacitation of bovine spermatozoa, part 3: Compositional and molecular changes to the plasma membrane.
Cryobiology
December 2024
Institute of Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76, Nitra, Slovakia.
Article Synopsis
- The study investigated how cryopreservation affects bovine sperm by measuring important proteins related to capacitation and apoptosis alongside membrane composition.
- Results showed that cryopreserved sperm (CRYO) had significantly lower sperm movement and higher DNA damage compared to both capacitated (CAP) and non-capacitated (CRTL) sperm.
- Notable changes in membrane composition included reduced levels of docosahexaenoic acid and cholesterol in CRYO sperm, along with down-regulated expression of capacitation-related proteins and altered levels of apoptosis-related proteins.
Effect of tyrode albumin lactate pyruvate and modified Krebs Ringers broth media on in vitro capacitation of HD-K75 boar spermatozoa at different period of incubation.
Reprod Domest Anim
August 2024
College of Veterinary Science, Assam Agricultural University, Guwahati, Assam, India.
In vitro capacitation allows for a greater understanding of the mechanisms underlying fertilization and the development of improved reproductive techniques for improving fertility rates in porcine. Tyrodes albumin lactate pyruvate (TALP) and modified Krebs Ringers Broth (m-KRB) are two medias that are commonly used in research experiments to induce capacitation in boar spermatozoa (Cañón-Beltrán et al., Theriogenology, 198, 2023 and 231; Oberlender et al.
View Article and Find Full Text PDFInduction of acrosome reaction by 4-Br-A23187 alters the glycoproteomic profile of boar spermatozoa.
Theriogenology
July 2024
Departamento de Bioquímica y Biología Molecular y Genética, Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain. Electronic address:
Protein glycosylation is a post-translational modification involved in wide range of biological processes. In mammalian spermatozoa this modification has been identified in numerous proteins, and membrane glycoproteins are involved in the fertilization process. The objective of the present study was to identify changes in protein glycosylation after acrosome reaction (AR) induction using the 4-Br-A23187 ionophore.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!