Abstract

BackgroundMicrotubule organization is essential for bipolar spindle assembly and chromosome segregation, which contribute to genome stability. Kinesin-5 Eg5 is known to be a crucial regulator in centrosome separation and spindle assembly in mammalian somatic cells, however, the functions and mechanisms of Eg5 in male meiotic cell division remain largely unknown.ResultsIn this study, we have found that Eg5 proteins are expressed in mouse spermatogonia, spermatocytes and spermatids. After Eg5 inhibition by specific inhibitors Monastrol, STLC and Dimethylenastron, the meiotic spindles of dividing spermatocytes show spindle collapse and the defects in bipolar spindle formation. We demonstrate that Eg5 regulates spindle bipolarity and the maintenance of meiotic spindles in meiosis. Eg5 inhibition leads to monopolar spindles, spindle abnormalities and chromosome misalignment in cultured GC-2 spd cells. Furthermore, Eg5 inhibition results in the decrease of the spermatids and the abnormalities in mature sperms.ConclusionsOur results have revealed an important role of kinesin-5 Eg5 in male meiosis and the maintenance of male fertility. We demonstrate that Eg5 is crucial for bipolar spindle assembly and chromosome alignment in dividing spermatocytes. Our data provide insights into the functions of Eg5 in meiotic spindle assembly of dividing spermatocytes.

Highlights

  • Microtubule organization is essential for bipolar spindle assembly and chromosome segregation, which contribute to genome stability

  • The inhibition of Eg5 leads to the monopolar spindles, spindle abnormalities and chromosome misalignment

  • Eg5 proteins locate at the cytoplasm and spindle microtubules in mouse spermatogenic cells We first used the immunofluorescence assay to investigate the distribution of Eg5 proteins in mouse testes

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Summary

Introduction

Microtubule organization is essential for bipolar spindle assembly and chromosome segregation, which contribute to genome stability. Microtubule-associated proteins, such as kinesins and dynein, are required for spindle assembly and chromosome movements during cell division [2]. Kinesin-5 motors are evolutionarily conserved motor proteins with plus-end-directed motility [3, 4]. Kinesin-5 Eg5 crosslinks parallel microtubules and slides antiparallel microtubules to stabilize spindle microtubules in mitosis [5, 9, 10]. Antiparallel interpolar microtubules are pushed apart by Eg5 through its plus-end-directed motility [11]. Recent studies have shown that kinesin-5 acts as a molecular brake within antiparallel microtubules to regulate microtubule motions [8, 16, 17]

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