Oogenesis is known to be important for embryonic pattern formation. For this reason we have studied the early differentiation of the honeybee ovariole histologically, ultrastructurally, and by staining F-actin with rhodaminyl-phalloidin. At the anterior tip of the ovariole, stem cells are lined up in a single file; they are organelle-poor but contain characteristic electrondense bodies with lysosomal properties. The presence of these bodies in cystocytes as well as prefollicle cells indicates that both cell types may be derived from the apical stem cells. During later stages of oogenesis, the follicle cells differentiate cytologically in different regions of the follicle. The organization of the intercellular bridges between cystocytes derived from a single cystoblast has been studied in detail. The polyfusomes in the intercellular bridges of cystocyte clusters stain with rhodaminyl-phalloidin and hence contain F-actin. Later, when the polyfusomes begin to desintegrate, F-actin rings form which line the rims of the intercellular bridges. Actin might be recruited from conspicuous F-actin stores which were detected in the germ-line cells. The F-actin rings are dissembled some time before the onset of vitellogenesis when the nurse chamber has grown to a length of about 200 μm. At the basal side of the follicle cells (close to the basement membrane facing the haemocdele) parallel microfilament bundles encircle the ovariole. The microfilament bundles which are oriented mostly perpendicular to the long axis of the ovariole were first observed around the zone where the cystocyte divisions occur; after this phase the micro-filament bundles become organized differently in the follicle cells associated with the nurse cells and in the follicular epithelium of the oocyte.

Cytoplasmic streaming in follicles of Drosophila has been analysed in vitro by means of time-lapse films. Late vitellogenic follicles develop normally in vitro as judged by morphological criteria. Furthermore, follicles (stage 10 and younger) which were cultured in vitro for the same length of time as follicles which were filmed, developed normally in vivo after injection into a host fly. The recorded cytoplasmic movements are, therefore, unlikely to be an in vitro artefact. At early vitellogenic stages (up to stage 9; King, 1970) no cytoplasmic streaming can be detected, but at stage 10A cytoplasmic movements are initiated within the oocyte. At stage 10B, when the nurse cells start degenerating, nurse cell cytoplasm can be seen to flow into the growing oocyte. At stage 11 a central stream of nurse-cell cytoplasm reaches the oocyte within a minute. The ooplasmic streaming is most rapid at stage 10B and stage 11 and only an oocyte cortex up to 7 μm thick remains stationary. Once the bulk of the nurse-cell cytoplasm has poured into the oocyte (stage 12) the cytoplasmic movement ceases, first in the nurse cells and later in the ooplasm. In mature oocytes no cytoplasmic streaming can be detected.

During the last phase of oogenesis in Drosophila, nurse cell cytoplasm can be seen to be streaming into the growing oocyte when visualized in time-lapse films. This process can be reversibly inhibited by cytochalasins. The distribution of F-actin filaments in the nurse cells has been studied by staining with rhodamine-conjugated phalloidin. At the beginning of cytoplasmic streaming (stage 10B) increasingly thick bundles of microfilaments formed, many of which spanned the nurse cell cytoplasm from the cell membrane to the nuclear membrane. The association of F-actin with the nuclear membrane persisted when nurse cell nuclei were isolated mechanically. The experimental evidence suggests that microfilament contraction in the nurse cells leads to cytoplasmic streaming by pressure flow.

No abstract