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During early oogenesis in Dysdercus fasciatus, anteriorly positioned nurse cells supply each oocyte with mRNA, ribosomes, and proteins via a microtubule-rich nutritive tube that lengthens as the oocyte is displaced backwards down an ovariole. Nurse cell-dependent development of an oocyte continues until the latter reaches a particular stage of oogenesis after which the nutritive tube supplying it becomes redundant and breaks down. The signal for nutritive tube breakdown is believed to derive from the oocyte, and to be developmental stage-specific. To explore this, nutritive tube turnover has been investigated following the experimental inhibition of oocyte maturation both by the prevention of mating, and also the topical application of precocene II. In each case, the nutritive tubes with their component microtubules continued to extend and failed to show normal tube redundancy, typified by microtubule rearrangement and then depolymerisation. This provided an in vivo demonstration that the dynamics of a large microtubule aggregate are influenced by the developmental state of the cytoplasm., (Copyright 2002 Wiley-Liss, Inc.)

Nutritive tubes that link the developing oocytes to the nurse cells in ovarioles of hemipteran insects contain extensive arrays of microtubules. These are established, then later depolymerised, by developmentally regulated processes. Breakdown of the microtubules corresponds with the activation of M-phase promoting factor (MPF) and mitogen-activated protein kinase (MAP kinase), later in oogenesis, as the oocytes proceed to arrest at the first meiotic metaphase [Lane and Stebbings, Roux's Arch Dev Biol 205:150-159 (1995)]. The mechanisms that lead to the breakdown of nutritive tube microtubules are unknown. Here, we have investigated the possibility that the insect ovarian microtubules are regulated by MPF- or MAP kinase-dependent phosphorylation, focusing upon the prominent high molecular weight microtubule-associated protein (HMW MAP) enriched in this system, which is a potential target for protein kinase activity in vivo. We have purified the prominent HMW MAPs from the ovaries of two species of hemipterans, and have shown them to be substrates in vitro for the activities of MPF and MAP kinase. However, although the catalytic component of MPF (p34cdc2) is present within microtubule-rich portions of hemipteran ovarioles, we have found that neither this protein nor its regulatory partner (cyclin B) co-purify with microtubules during taxol-mediated microtubule isolation.