Molecular underpinnings of the early brain developmental response to differential feeding in the honey bee Apis mellifera.

Brain differential morphogenesis in females is one of the major phenotypic manifestations of caste development in honey bees. Brain diphenism appears at the fourth larval phase as a result of the differential feeding regime developing females are submitted during early phases of larval development. Here, we used a forward genetics approach to test the early brain molecular response to differential feeding leading to the brain diphenism observed at later developmental phases. Using RNA sequencing analysis, we identified 53 differentially expressed genes (DEGs) between the brains of queens and workers at the third larval phase. Since miRNAs have been suggested to play a role in caste differentiation after horizontal and vertical transmission, we tested their potential participation in regulating the DEGs. The miRNA-mRNA interaction network, including the DEGs and the royal- and worker-jelly enriched miRNA populations, revealed a subset of miRNAs potentially involved in regulating the expression of DEGs. The interaction of miR-34, miR-210, and miR-317 with Takeout, Neurotrophin-1, Forked, and Masquerade genes was experimentally confirmed using a luciferase reporter system. Taken together, our results reconstruct the regulatory network that governs the development of the early brain diphenism in honey bees.

Effects of treatment of the fat body trophocytes of Melipona quadrifasciata anthidioides nurse workers and virgin queens in culture by juvenile hormone III and ecdysterone (20-HE).

The fat body (FB) consists of two types of cells: throphocytes and oenocytes. Throphocytes are related to intermediary metabolism storing lipids, carbohydrates, and proteins while oenocytes play role in the lipids and lipoproteins production. The vitellogenin is the precursor of egg yolk (vitelline) and is synthesized on FB. The aim of this work was to analyze the effects of hormones acting in bee reproduction, as juvenile hormone (JH) and ecdisteroids (20 HE) on FB cells, where vitellogenin is synthesized. For the study were chose nurse workers that in Melipona quadrifasciata anthidioides present activated ovaries and produce eggs, and virgin queens whose ovaries are not yet activated, presenting only previtellogenic follicles. FB trophocytes from these classes of bees were cultivated in media containing different amounts of JH and 20-HE. The effects on trophocytes cytoplasm reserves of lipids, proteins, and activity of acid phosphatase were compared by observing preparations from cultured FB, treated and control, by transmission electron microscopy (TEM). The results showed that the hormones effects are related to the bee's caste and functional ovary stage. The role of acid phosphatase on mobilization of the trophocyte reserves was also determined.

The stomatogastric nervous system of the honey bee (Apis mellifera) in a critical phase of caste development.

Progress in our understanding of polymorphic differentiation of female honey bee larvae into queens and workers required a re-evaluation of neuronal pathways potentially involved in transmitting information on food quality. This study presents new data on the anatomy of one of these pathways, the stomatogastric nervous system (SNS) of honey bee larvae and pupae. Scanning electron microscopy preparations demonstrated not only developmental changes in frontal ganglion structure, but also provided firm evidence for a hypocerebral ganglion in the honey SNS. In addition to previously described SNS nerves, the frontal, recurrent and esophageal nerves, and the frontal connectives, we observed three new nerves that connect the SNS to the central nervous system and the foregut. The first one is an unpaired connective nerve of the frontal ganglion to the anteromedial protocerebrum. The second consists of paired lateral branches of the recurrent nerve, and the third is a plexus of fine nervous branches associated with the pharynx. Lateral extensions of the newly described hypocerebral ganglion also make contact with the pharynx. Similar but smaller branches were also observed to originate from the esophageal nerves as they run along the foregut. The exact anatomical localization of the cardiostomatogastric nerves, which connect the SNS with the retrocerebral complex, could also be detected. The description of such new nervous connections will serve as a database for functional analyses on the role of the SNS in differential feeding responses of the honey bee larvae, representing the initial step in caste differentiation. J. Morphol. 236:139-149, 1998. © 1998 Wiley-Liss, Inc.