Motoneuron axon pathfinding errors in zebrafish: differential effects related to concentration and timing of nicotine exposure.

Nicotine exposure during embryonic stages of development can affect many neurodevelopmental processes. In the developing zebrafish, exposure to nicotine was reported to cause axonal pathfinding errors in the later born secondary motoneurons (SMNs). These alterations in SMN axon morphology coincided with muscle degeneration at high nicotine concentrations (15-30 µM). Previous work showed that the paralytic mutant zebrafish known as sofa potato exhibited nicotine-induced effects onto SMN axons at these high concentrations but in the absence of any muscle deficits, indicating that pathfinding errors could occur independent of muscle effects. In this study, we used varying concentrations of nicotine at different developmental windows of exposure to specifically isolate its effects onto subpopulations of motoneuron axons. We found that nicotine exposure can affect SMN axon morphology in a dose-dependent manner. At low concentrations of nicotine, SMN axons exhibited pathfinding errors, in the absence of any nicotine-induced muscle abnormalities. Moreover, the nicotine exposure paradigms used affected the 3 subpopulations of SMN axons differently, but the dorsal projecting SMN axons were primarily affected. We then identified morphologically distinct pathfinding errors that best described the nicotine-induced effects on dorsal projecting SMN axons. To test whether SMN pathfinding was potentially influenced by alterations in the early born primary motoneuron (PMN), we performed dual labeling studies, where both PMN and SMN axons were simultaneously labeled with antibodies. We show that only a subset of the SMN axon pathfinding errors coincided with abnormal PMN axonal targeting in nicotine-exposed zebrafish. We conclude that nicotine exposure can exert differential effects depending on the levels of nicotine and developmental exposure window.

Evaluation of Moringa oleifera as a dietary supplement on growth and reproductive performance in zebrafish.

The leaves of the Moringa oleifera (Moringa) tree contain a significant source of protein, vitamins and minerals, and are considered as an important dietary supplement in countries where chronic malnourishment is linked to poor fetal development. We evaluated the effectiveness of the Moringa leaf as a supplemental replacement for vitamins, minerals, and protein in a formulated zebrafish diet and the impact that it may have on growth and reproductive outcome. Diets included a formulated control (FC) containing an array of vitamins and mineral supplements (pre-mixes), dried ground Moringa only (M), formulated control minus vitamin and mineral pre-mixes (Fvm), and formulated control minus vitamin and mineral pre-mixes and supplemented with Moringa (FM). Juvenile zebrafish were fed experimental diets ad libitum. After a 12 week feeding period, each treatment group was evaluated based on growth and reproductive performance. The M treatment showed the least growth performance (length and weight gain) and no reproductive success (no egg production). Although small, M fish appeared otherwise healthy, with survivorship at ca. 70%, suggesting, Moringa can serve as a single ingredient source for a short period of time. FC showed the highest growth performance, and had the highest reproductive success. Growth performance and reproduction in the Fvm diet was greatly reduced. However, inclusion of Moringa (FM) promoted significant, but not total, recovery of growth and reproductive metrics. These data suggest that Moringa leaves can serve as an acceptable supplement for macro and micronutrients in the diet and could, in part, reduce problems associated with nutrient deficiencies.