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Eukaryotic cells have different motor proteins for transporting various cargo within the cell. These motor proteins differ based on the filament they associate with, the direction they move within the cell, and the type of cargo they transport. Motor proteins that associate with microtubules are known as microtubule-associated motor proteins. There are two families of microtubule-associated motor proteins —Kinesins and Dyneins. Both these proteins assist in the transport of cellular...
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The journey of sperm from its origin to the point of ejaculation begins within the seminiferous tubules of the testis. Here, Sertoli cells produce fluid that propels non-motile sperm through a series of conduits, starting with the straight tubules leading to the rete testis. This interconnected network of tubules acts as the initial pathway for sperm, guiding them into the efferent ductules and then into the epididymis for maturation.
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Microtubules are thick hollow cylindrical proteins that help form the cytoskeleton. Microtubules have varied roles in the cell. These filaments help form cellular appendages like cilia and flagella, which are responsible for locomotion. The cilia arise from basal bodies, separated from the main body by a membrane-like structure forming the transition zone. This zone is the gate for the entry of lipids and proteins, creating a unique composition of lipids and proteins in the ciliary membrane and...
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Related Experiment Video

Updated: Jan 7, 2026

Phosphopeptide Analysis of Rodent Epididymal Spermatozoa
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Autophagy-Related Proteins Influence Mouse Epididymal Sperm Motility.

Lorena Rodríguez-Páez1, Jonathan J Magaña2,3, Charmina Aguirre-Alvarado1,4

  • 1Laboratorio de Bioquímica Farmacológica, Departamento de Bioquímica, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prol. Carpio y Plan de Ayala, Col. Santo Tomás, Mexico City 11340, Mexico.

International Journal of Molecular Sciences
|December 30, 2025
PubMed
Summary

Mouse sperm may initiate autophagy, a cellular recycling process, during capacitation. Autophagy proteins LC3, p62/SQSTM1, and mTOR dynamically relocate, impacting sperm motility and acrosome reaction.

Keywords:
LC3acrosome reactionmTORmotilityp62/SQSTM1spermatozoa

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Using an Extracellular Flux Analyzer to Measure Changes in Glycolysis and Oxidative Phosphorylation during Mouse Sperm Capacitation
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Area of Science:

  • Cell Biology
  • Reproductive Biology
  • Autophagy Research

Background:

  • Autophagy is a vital cellular process for recycling components and maintaining homeostasis.
  • The role and mechanisms of autophagy in mammalian sperm, particularly mouse epididymal sperm, remain largely unexplored.
  • Key autophagy markers like LC3, p62/SQSTM1, and mTOR are crucial for understanding autophagic flux.

Purpose of the Study:

  • To investigate the presence and dynamic changes of autophagy markers (LC3, p62/SQSTM1, mTOR) in mouse epididymal sperm during capacitation.
  • To evaluate the functional significance of these autophagy-related proteins on sperm physiology, including viability, motility, and acrosome reaction.
  • To assess the impact of autophagy modulation using inhibitors (chloroquine, K67) and activators (rapamycin) on sperm function.

Main Methods:

  • Western blotting to detect LC3, p62/SQSTM1, and mTOR protein levels in epididymal mouse sperm.
  • Analysis of sperm parameters: viability, motility, intracellular pH, intracellular calcium, mitochondrial membrane potential, and acrosome reaction induction.
  • Pharmacological treatments using chloroquine (CQ), K67, and rapamycin to modulate autophagy pathways.

Main Results:

  • Autophagy markers LC3, p62/SQSTM1, and mTOR exhibited dynamic re-localization in mouse sperm under capacitation conditions.
  • Autophagy inhibitors (CQ, K67) decreased LC3-II and p62/SQSTM1 levels, while rapamycin did not increase mTOR levels.
  • Inhibitor treatment led to reduced sperm motility, decreased mitochondrial membrane potential, and impaired acrosome reaction induction, without affecting sperm viability.

Conclusions:

  • Mouse epididymal sperm possess the machinery for initial autophagy steps, evidenced by the dynamic presence and re-localization of key autophagy proteins during capacitation.
  • Autophagy-related proteins play a significant role in regulating sperm motility and acrosome reaction induction.
  • Pharmacological inhibition of autophagy negatively impacts critical sperm functions, highlighting autophagy's importance in male fertility.