Stage-specific appearance of cytoplasmic microtubules around the surviving nuclei during the third prezygotic division of Paramecium

There are six micronuclear divisions during conjugation of Paramecium caudatum:three prezygotic and three postzygotic divisions.Four haploid nuclei are formed during the first two meiotic prezygotic divisions.Usually only one meiotic product is located in the paroral cone(PC)region at the completion of meiosis,which survives and divides mitotically to complete the third prezygotic division to yield a stationary and a migratory pronucleus.The remaining three located outside of the PC degenerate.The migratory pronuclei are then exchanged between two conjugants and fuse with the stationary pronuclei to form synkarya,which undergo three successive divisions(postzygotic divisions).However,little is known about the surviving mechanism of the PC nuclei.In the current study,stage-specific appearance of cytoplasmic microtubules(cMTs)was indicated during the third prezygotic division by immunofluorescence labeling with anti-alpha tubulin antibodies surrounding the surviving nuclei,including the PC nuclei and the two types of prospective pronuclei.This suggested that cMTs were involved in the formation of a physical barrier,whose function may relate to sequestering and protecting the surviving nuclei from the major cytoplasm,where degeneration of extra-meiotic products occurs,another important nuclear event during the third prezygotic division.