Ultrastructure of light-activated axons following optogenetic stimulation to produce late-phase long-term potentiation.

Analysis of neuronal compartments has revealed many state-dependent changes in geometry but establishing synapse-specific mechanisms at the nanoscale has proven elusive. We co-expressed channelrhodopsin2-GFP and mAPEX2 in a subset of hippocampal CA3 neurons and used trains of light to induce late-phase long-term potentiation (L-LTP) in area CA1. L-LTP was shown to be specific to the labeled axons by severing CA3 inputs, which prevented back-propagating recruitment of unlabeled axons. Membrane-associated mAPEX2 tolerated microwave-enhanced chemical fixation and drove tyramide signal amplification to deposit Alexa Fluor dyes in the light-activated axons. Subsequent post-embedding immunogold labeling resulted in outstanding ultrastructure and clear distinctions between labeled (activated), and unlabeled axons without obscuring subcellular organelles. The gold-labeled axons in potentiated slices were reconstructed through serial section electron microscopy; presynaptic vesicles and other constituents could be quantified unambiguously. The genetic specification, reliable physiology, and compatibility with established methods for ultrastructural preservation make this an ideal approach to link synapse ultrastructure and function in intact circuits.

Hazards inherent in interdisciplinary behavioral research.

Many, if not all, questions in biology and psychology today were formulated and considered in depth, though typically in a different language, from the 1700's to the early 1900's. However, because of politics or fashion, some topics fell out of favor or failed to recruit new scientists and hence languished. Despite greatly expanded scholarship in the history of the life sciences in the twentieth century, many such topics have had to be rediscovered in recent years, while much of the wisdom already accrued stays in the older literature and not in active minds. This is particularly true today when scientific advances appear at breakneck speed. It would not be an exaggeration to say that many 'breakthroughs' turn out really to be rediscoveries of forgotten observations. Two areas of particular significance to the interdisciplinary study of behavior are the Norms of Reaction (from Biology) and the concept of Plasticity (from Psychology). These and related fields benefit from the perspective of epigenetics so long as rigorous operational definitions are implemented. It is also important to revive Hogben's admonition that the interaction of hereditary and environment cannot be understood outside of the context of development. Five examples of increasing complexity in phenotypic plasticity in brain and behavior are presented to illustrate this perspective.