The activity patterns of Individual neurons are highly coordinated and synchronized within neuronal circuits in the brain, much like individual orchestra tools playing together to achieve harmony. Inside neurons, complex protein signaling cascades provide the molecular notes and instructions to each neuron. However, until recently, the dynamic nature of intracellular protein signaling in the intact brain has been eluded. In this review, we focus on recent advancements and the development of approaches to study neuronal signaling dynamics in vivo. We will discuss approaches for the implementation of biosensors for monitoring of protein signaling activities at the levels of individual synapses, dendritic branches, cell-wide neuromodulation, and transcription in the nucleus. Future improvement in these methods and their utilization will undoubtedly yield new insights regarding the intricate link between functional and molecular neuronal dynamics and how they underlie animal’s behavior.