Clathrin is essential for meiotic spindle function in oocytes.
In the mammalian ovary, oocytes are arrested at prophase of meiosis I until a hormonal stimulus triggers resumption of meiosis. During the subsequent meiotic maturation process, which includes completion of the first meiotic division and formation of the second metaphase spindle, oocytes acquire competence for fertilization. Recently, it was shown that clathrin, a cytosolic protein complex originally defined for its role in intracellular membrane traffic, is also involved in the stabilization of kinetochore fibers in mitotic spindles of dividing somatic cells. However, whether clathrin has a similar function in meiotic spindles in oocytes has not been investigated previously. Our results show that endogenous clathrin associates with the meiotic spindles in oocytes. To study the function of clathrin during meiotic maturation, we microinjected green fluorescent protein-tagged C-terminal and N-terminal dominant-negative clathrin protein constructs into isolated porcine oocytes prior to in vitro maturation. Both protein constructs associated with meiotic spindles similar to endogenous clathrin, but induced misalignment and clumping of chromosomes, occurrence of cytoplasmic chromatin and failure of polar body extrusion. These data demonstrate that clathrin plays a crucial role in meiotic spindle function in maturing oocytes, possibly through spindle stabilization.
Departments of Farm Animal Health Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Jurriaan J
JJ
Hölzenspies
Bernard A J
BA
Roelen
Ben
B
Colenbrander
Roland A P
RA
Romijn
Wieger
W
Hemrika
Willem
W
Stoorvogel
Theo
T
van Haeften
D000818:Animals; D002966:Clathrin / Q000502:physiology*; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D049452:Green Fluorescent Proteins / Q000235:genetics; D008540:Meiosis / Q000502:physiology*; D008845:Microinjections / Q000379:methods; D018613:Microscopy, Confocal; D009865:Oocytes / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D012044:Regression Analysis; D008941:Spindle Apparatus / Q000502:physiology*; D013552:Swine
D002966:Clathrin; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins