Standard methods for physiology and biochemistry research in Apis mellifera

Despite their tremendous economic importance, and apart from certain topics in the field of neurophysiology such as vision, olfaction, learning and memory, honey bees are not a typical model system for studying general questions of insect physiology. The reason is their social lifestyle, which sets them apart from a typical insect and, during social evolution, has resulted in the restructuring of certain physiological pathways and biochemical characteristics in this insect. Not surprisingly, the questions that have attracted most attention by researchers working on honey bee physiology and biochemistry in general are core topics specifically related to social organization, such as caste development, reproductive division of labour and polyethism within the worker caste. With certain proteins playing key roles in these processes, such as the major royal jelly proteins (MRJPs), including royalactin and hexamerins in caste development, and vitellogenin in reproductive division of labour and age polyethism, a major section herein will present and discuss basic laboratory protocols for protein analyses established and standardized to address such questions in bees. A second major topic concerns endocrine mechanisms underlying processes of queen and worker development, as well as reproduction and polyethism, especially the roles of juvenile hormone and ecdysteroids. Sensitive techniques for the quantification of juvenile hormone levels circulating in haemolymph, as well as its synthesis by the corpora allata are described. Although these require certain instrumentation and a considerable degree of sophistication in the analysis procedures, we considered that presenting these techniques would be of interest to laboratories planning to specialize in such analyses. Since biogenic amines are both neurotransmitters and regulators of endocrine glands, we also present a standard method for the detection and analysis of certain biogenic amines of interest. Further questions that cross borders between individual and social physiology are related to energy metabolism and thermoregulation. Thus a further three sections are dedicated to protocols on carbohydrate quantification in body fluid, body temperature measurement and respirometry.