Neural circuits mediating olfactory-driven behavior in fish

Tyrosine hydroxylase immunohistochemistry is used to demonstrate catecholaminergic neuronal populations in the fore- and midbrain of adult zebrafish (Danio rerio). While no catecholaminergic neurons are found in the midbrain, various immunoreactive populations were found in the diencephalon (hypothalamus, posterior tuberculum, ventral thalamus, pretectum) and telencephalon (preoptic region, subpallium, olfactory bulb). The posterior tubercular catecholaminergic cells include three cytological types (small round, large pear-shaped, and bipolar liquor-contacting cells). Furthermore, the retrograde neuronal tracers DiI or biocytin were applied to demonstrate ascending projections to the basal telencephalon (incl. the striatum). A double-label approach was used - together with tyrosine hydroxylase immunohistochemistry - in order to visualize neurons positive for tyrosine hydroxylase and a retrograde tracer. Double-labeled cells were identified in two locations in the posterior tuberculum (i.e, small round neurons in the periventricular nucleus of the posterior tuberculum and large pear-shaped cells adjacent to it). They are interpreted as the teleostean dopaminergic system ascending to the striatum, since previous work [16] established that no noradrenergic neurons exist in the forebrain of the adult zebrafish.

Zebrafish (zebra danio) are becoming increasingly popular in behavioral neuroscience and behavior genetics. This small vertebrate may be utilized in modeling human brain disorders. One of the major neuropsychiatric conditions still not well understood is abnormally increased fear and anxiety. Zebrafish may be an appropriate organism with which these human diseases can be modeled and their biological mechanisms investigated. Predator induced anxiety paradigms have been suggested as useful methods in translational research. Shoaling fish, such as zebrafish, are known to respond to alarm substances with antipredatory or alarm reactions. However, these responses are not well characterized in zebrafish. In the current paper, we investigate the behavioral responses of zebrafish elicited by its alarm substance. Using observation-based as well as video-tracking aided behavior quantification methods we demonstrate significant alarm substance-induced behavioral changes that are independent of the presence of a predatory fish stimulus. The results suggest that, once refined, the use of alarm substance with zebrafish will allow the development of high throughput behavioral paradigms for drug and mutation screening aimed at the analysis of the biological mechanisms of fear in vertebrates.

Odors are thought to be represented by a distributed code across the glomerular modules in the olfactory bulb (OB). Here, we optically imaged presynaptic activity in glomerular modules of the zebrafish OB induced by a class of natural odorants (amino acids [AAs]) after labeling of primary afferents with a calcium-sensitive dye. AAs induce complex combinatorial patterns of active glomerular modules that are unique for different stimuli and concentrations. Quantitative analysis shows that defined molecular features of stimuli are correlated with activity in spatially confined groups of glomerular modules. These results provide direct evidence that identity and concentration of odorants are encoded by glomerular activity patterns and reveal a coarse chemotopic organization of the array of glomerular modules.