Chimeric honeybees (Apis mellifera) produced by transplantation of embryonic cells into pre‐gastrula stage embryos and detection of chimerism by use of microsatellite markers

Abstract

The production of chimeras, by use of cell transplantation, has proved to be highly valuable in studies of development by providing insights into cell fate, differentiation, and developmental potential. So far, chimeric honeybees have been created by nuclear transfer technologies. We have developed protocols to produce chimeric honeybees by use of cell transplantation. Embryonic cells were transplanted between pre-gastrula stage embryos (32-34 hr after oviposition) and hatched larvae were reared in vitro for 4 days. Chimeric individuals were detected by use of microsatellite analysis and a conservative estimation approach. 4.8% of embryos, posteriorly injected with embryonic cells, developed into chimeric honeybee larvae. By injection of cells pre-stained with fluorescent cell tracer dye, we studied the integration of transplanted cells in the developing embryos. Number of injected cells varied from 0 to 50 and cells remained and multiplied mainly in the area of injection.

Proteome Analysis Unravels Mechanism Underling the Embryogenesis of the Honeybee Drone and Its Divergence with the Worker (Apis mellifera lingustica)

“…Embryogenesis is the first stage in honeybee life circle, during which the rudimentary organs of adult bees are formed. , Accumulating evidence demonstrates that the embryo is the ideal model for honeybee genetic modification. Several physiological characters of the honeybee embryo make it an ideal stage of development for study, such as a thin chorion, a relatively easy maintenance process of the ambient environment (34 °C temperature, 80% relative humidity) during development, and the ability to easily puncture a hole on the shell and still have it develop normally under lab conditions. , Because of these biological merits, genetic manipulations of the honeybee embryo have been documented in the areas of transplantation of nuclear materials and RNA interference. − Recently, the in vitro cultivation of the embryonic cell − has offered a potential venue to study one target gene or protein that regulates the embryonic development of the honeybee.…”

In-depth Proteomics Characterization of Embryogenesis of the Honey Bee Worker (Apis mellifera ligustica)

“…For example, embryonic cells in the pre-gastrula stage that have been transplanted with nuclear materials have developed into chimeric honey bee larvae (15). RNA interference (RNAi) has been used for honey bee embryos in vivo to characterize the functioning of specific genes (16) and for genetic effects on morphological differentiation (17,18).…”

Long-term maintenance of in vitro cultured honeybee (Apis mellifera) embryonic cells

“…Our initial motivation for studying embryonic cells and for developing protocols for their culture was mainly to create a system allowing maintenance and manipulation of cells for subsequent injections into honeybee embryos. We have recently produced chimeric honeybees by cell transplantation between embryos [ 23 ]. Establishment of protocols for long-term cultivation of embryonic cells proves new opportunities for study of chimerism in bees.…”

“…Another application of cells in culture is as donors in cell transplantations for cell-mediated gene transfer and production of chimeras. Our group has recently successfully produced chimeras by transplantation of cells between embryos [ 23 ]. Access to long-term cultures of highly potential embryonic cells will give new opportunities for studies and transplantation of labelled and modified cells.…”