Immunolocalization of Neurotransmitters and Neuromodulators in the Developing Crayfish Brain

In the field of neurosciences, the crayfish nervous system is an important model for understanding how arthropods process sensory stimuli and generate specific behaviors. Furthermore, crayfish embryos have been important study objects for well over 200 years. Immunohistochemistry against neurotransmitters, neuromodulators, and neurohormones is widely used to analyze the ontogeny of neurons in the emerging brain of several crustacean species and to date represents one of the most powerful approaches to analyze aspects of brain development in this group of organisms. In recent years, the analysis of brain development in crustaceans has gained new momentum by the establishment of the Marmorkrebs Procambarus virginalis (Marbled Crayfish), a parthenogenetic crayfish, as new model system. The embryonic development of marbled crayfish is well characterized and these animals can be easily cultivated in the lab. This chapter describes protocols for immunolocalization of neuroactive substances in the developing crayfish brain.