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

BACKGROUND: In vitro cultivation of cells allows novel investigation of in vivo- mechanisms and is a helpful tool in developmental biology, biochemistry and functional genomics. Numerous cell lines of insect species, e.g., silkworm and mosquito, have been reported. However, this is not the case for successful long-term cultivation of cells in honeybees. RESULTS: Methods for cultivation of honeybee embryonic cells are discussed here. Pre-gastrula stage embryos were used to initiate cultures, and cells were reared on 96-wells microplates with Grace insect medium, supplemented with Fetal Bovine Serum. Cells proliferated in clusters, and maintained viable and mitotic for more than three months. CONCLUSION: We report here, for the first time, long-term cultivation of honeybee cells. Results represent a highly useful in vitro-system for studying a model organism of increasing importance in areas such as aging, sociality and neurobiology

Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA

BACKGROUND: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. Here we assess two methods for probing gene function by RNA interference (RNAi) in adult honeybees. RESULTS: The vitellogenin gene was chosen as target because its expression is unlikely to have a phenotypic effect until the adult stage in bees. This allowed us to introduce dsRNA in preblastoderm eggs without affecting gene function during development. Of workers reared from eggs injected with dsRNA derived from a 504 bp stretch of the vitellogenin coding sequence, 15% had strongly reduced levels of vitellogenin mRNA. When dsRNA was introduced by intra-abdominal injection in newly emerged bees, almost all individuals (96 %) showed the mutant phenotype. An RNA-fragment with an apparent size similar to the template dsRNA was still present in this group after 15 days. CONCLUSION: Injection of dsRNA in eggs at the preblastoderm stage seems to allow disruption of gene function in all developmental stages. To dissect gene function in the adult stage, the intra-abdominal injection technique seems superior to egg injection as it gives a much higher penetrance, it is much simpler, and it makes it possible to address genes that are also expressed in the embryonic, larval or pupal stages

Immunogold Localization of Vitellogenin in the Ovaries, Hypopharyngeal Glands and Head Fat Bodies of Honeybee Workers, Apis Mellifera

Vitellogenin is a yolk precursor protein in most oviparous females. In the advanced eusocial honeybee, Apis mellifera (Hymenoptera: Apidae), vitellogenin has recently attracted much interest as this protein, in addition to a classical function in oocyte development in the reproductive queen caste, has evolved functions in the facultatively sterile female worker caste not documented in other species. However, research on the spatial dynamics of vitellogenin in various tissues is not easily performed with available tools. Here we present an immunogold staining procedure that visualizes honeybee vitellogenin in resin embedded tissue. To establish the protocol, we used ovaries of worker bees from colonies with and without a queen. Under the first condition, vitellogenin is assumed not to be present in the workers' ovaries. Under the second condition, the ovaries of worker bees become vitellogenic, with abundant opportunities for detection of complex patterns of vitellogenin uptake and storage. By use of this experimental setup, the staining method is shown to be both sensitive and specific. To demonstrate the functional significance of the protocol, it was subsequently used to identify vitellogenin protein in the hypopharyngeal glands (brood food producing head glands) of nursing worker bees and in adjacent head fat body cells for the first time. Localization of vitellogenin in these tissues supports previously hypothesized roles of vitellogenin in social behavior. This protocol thus provides deeper insights into the functions of vitellogenin in the honeybee

Citizen Scientists Discover a New Auroral Form : Dunes Provide Insight Into the Upper Atmosphere

Auroral forms are like fingerprints linking optical features to physical phenomena in the near‐Earth space. While discovering new forms is rare, recently, scientists reported of citizens' observations of STEVE, a pinkish optical manifestation of subauroral ionospheric drifts that were not thought to be visible to the naked eye. Here, we present a new auroral form named “the dunes”. On 7 October 2018, citizen observers took multiple digital photographs of the same dunes simultaneously from different locations in Finland and Sweden. We develop a triangulation method to analyze the photographs and conclude that the dunes are a monochromatic wave field with a wavelength of about 45 km within a thin layer at 100 km altitude. Supporting data suggest that the dunes manifest atmospheric waves, possibly mesospheric bores, which are rarely detected, and have not previously been observed via diffuse aurora nor at auroral latitudes and altitudes. The dunes present a new opportunity to investigate the coupling of the lower/middle atmosphere to the thermosphere and ionosphere. Our paper adds to the growing body of work that illustrates the value of citizen scientist images in carrying out quantitative analysis of optical phenomena, especially at small scales at subauroral latitudes. Further, the dune project presents means to create general interest toward physics, emphasizing that citizens can take part in scientific work by helping to uncover new phenomena.Peer reviewe