Raising female-destined larvae into new queens by alloparental care of nurse bees in provision of royal jelly (RJ) is critical for reproductive investment of honeybees (Apis mellifera) by alloparental care, which requires more than one brain region involving in neural responses. However, knowledge of how mushroom bodies (MBs) and antennal lobes (ALs) of brain influence the alloparental care of honeybees is largely unknown. We compared the proteome of MBs and ALs in the brain of two strains of honeybees with different reproductive investment in terms of high (royal jelly bees selected from Italian bees > 40 years, RJbs) and low (Italian bees, ITbs) RJ production. In newly emerged bees, the induced pathways associated with energetic metabolism in MBs and ALs of RJbs comparing to ITbs, indicate that the developmental trajectory of brain has reshaped since eclosion. In nurse ALs of RJbs comparing to ITbs, the pathways and kinases related to modulate vesicular transport, signal transduction, synaptic plasticity, and long-term memory consolidation are functionally enhanced to boost the activity of neuronal cell to sense the brood odor. The difference in ALs is also found in forager bees between both bee lines, of which pathway related to protein metabolism are functionally activated, however there was no difference in MBs between them. Among of all the differences, of which the enhanced activity of nurse ALs plays the major role in driving the stronger reproductive investment in RJbs than in ITbs. Our findings indicate that RJbs has adapted a strong olfactory sense to larvae by alloparental care due to the genetic selection of increasing royal jelly outputs. This gains a novel insight into a neural biology of honeybees, and is potentially useful for neurophysiology for honeybees and other social insects.