Proteome And Phosphoproteome Comparison Of Honeybee Brain Sub-Region

Despite well-known cerebral functions of the brain sub-regions,knowledge on how proteome underpins specialized neural functions in different anatomical regions in honeybee and the differences between eastern and western honeybee species are still lacking.We characterized and compared the proteomes of mushroom bodies?MBs?,antennal lobes?ALs?and optical lobes?OLs?in the brain of both species,and biologically validated the functions related to learning and memory.Acc and Aml have evolved similar proteome signatures in MBs and OLs to drive the domain-specific neural activities.In MBs of both species,commonly enriched and enhanced functional groups related to protein metabolism and Ca²⁺transport relative to ALs and OLs,suggests that proteins and Ca²⁺are vital for consolidating learning and memory via modulation of synaptic structure and signal transduction.Furthermore,in OLs of both species,the mainly enriched ribonucleoside metabolism suggests its vital role as second messenger in promoting phototransduction.Notably,in ALs of both species,distinct proteome settings have shaped to prime olfactory learning and memory.In ALs of Acc,this is supported by the enriched cytoskeleton organization to sustain olfactory signaling through modulation of plasticity in glomeruli and intracellular transport.In ALs of Aml,however,the enriched functional groups implicated in hydrogen ion transport are indicative of their importance in supporting olfactory processes by regulation of synaptic transmission.The biological confirmation of enhanced activities of protein metabolism and signal transduction in ALs and MBs of Acc relative to in Aml demonstrates that a stronger sense of olfactory learning and memory has evolved in Acc.Region-specific physiology of honeybee brain requires phosphorylation-dependent intracellular signaling.To better understand the role of phosphorylation in maintenance of physiological difference among three main functional regions of honeybee brain,we performed phosphoprteomic characterizations of MBs,ALs and OLs using LC-MS/MS.Our data suggests that the‘‘typical''phosphoprotein is widely expressed yet displays variable phosphorylation site.The sub-region phosphorylation tunes protein activity to the specific functional of each region.In MBs,the phosphorylation promotes the learning and memory ability via enhance the phosphatidyl inositol signal transduction.In ALs,phosphoproteins mainly envolved in mTOR signaling pathway and phagosome facilitate the olfactory process.In OLs,the phosphorylation event mainly implicated in transport suggests that the phosphorylation play key roles in ion transport during phototransduction process.And STE/STE 20 kinase family may play important roles in phototransduction.Furthermore,when compared the phospohproteins expression of each sub-regions between two bee species,we found that the phosphorylaton event of MBs and OLs between two species were similar.Surprisingly,many key phosphorylation site in mTOR signanling pathway in ALs of Acc have up-regulated comparation to Aml,this indicates that mTOR signanling pathway may responsibility for Acc's better ability in foraging scattered nectar sources.The reported first in-depth proteome and phosphoproteome data of honeybee brain sub-organs provide a novel insight into the molecular basis of neurobiology,and is potentially useful for further neurological studies in honeybees and other insects.