Determination of relative age using growth increments of scales as a minimally invasive method in the tropical freshwater Apteronotus leptorhynchus.

This study describes a method for the determination of relative age in a tropical teleost, the brown ghost knifefish Apteronotus leptorhynchus. This method is based on identification of the maximum number of scale circuli, which is thought to be associated with the oldest scales, and thus to be the most indicative of the age of a given fish. Relative age can be inferred by relating differences in maximum circulus counts to the average rate of circulus addition, which was estimated at 34 circuli per year in adult fish through oxytetracycline marking. This method shows high inter-investigator reliability and has a limited effect on fish because of the low number of scales required in order to determine the maximum number of circuli with a sufficiently high confidence level. Analysis of the frequency distribution of the circulus counts revealed periodic patterns that are similar among fish, presumably reflecting the environmental life history of the individuals. Regression analysis and comparison of addition rates showed that scale circulus counts and otolith ring counts are equivalent approaches for age estimation, but scale analysis is superior because of its limited invasiveness and the lower demand in terms of technical skills and expensive instrumentation.

Proteome analysis reveals protein candidates involved in early stages of brain regeneration of teleost fish.

Exploration of the molecular dynamics underlying regeneration in the central nervous system of regeneration-competent organisms has received little attention thus far. By combining a cerebellar lesion paradigm with differential proteome analysis at a post-lesion survival time of 30 min, we screened for protein candidates involved in the early stages of regeneration in the cerebellum of such an organism, the teleost fish Apteronotus leptorhynchus. Out of 769 protein spots, the intensity of 26 spots was significantly increased by a factor of at least 1.5 in the lesioned hemisphere, relative to the intact hemisphere. The intensity of 9 protein spots was significantly reduced by a factor of at least 1.5. The proteins associated with 15 of the spots were identified by peptide mass fingerprinting and/or tandem mass spectrometry, resulting in the identification of a total of 11 proteins. Proteins whose abundance was significantly increased include: erythrocyte membrane protein 4.1N, fibrinogen gamma polypeptide, fructose-biphosphate aldolase C, alpha-internexin neuronal intermediate filament protein, major histocompatibility complex class I heavy chain, 26S proteasome non-ATPase regulatory subunit 8, tubulin alpha-1C chain, and ubiquitin-specific protease 5. Proteins with significantly decreased levels of abundance include: brain glycogen phosphorylase, neuron-specific calcium-binding protein hippocalcin, and spectrin alpha 2. We hypothesize that these proteins are involved in energy metabolism, blood clotting, electron transfer in oxidative reactions, cytoskeleton degradation, apoptotic cell death, synaptic plasticity, axonal regeneration, and promotion of mitotic activity.

Radial glia in the cerebellum of adult teleost fish: implications for the guidance of migrating new neurons.

In contrast to mammals, in teleost fish radial glia persist beyond early development. This persistence parallels the enormous potential of teleosts to continuously generate a large number of new neurons in dozens of specific proliferation zones in the adult brain. In the present study, we characterized in the teleost fish Apteronotus leptorhynchus the immunological properties of radial glia in the corpus cerebelli-a cerebellar subdivision with particularly high proliferative activity-and examined their possible function in the guidance of migrating young neurons. Radial glia stained immunopositive for glial fibrillary acidic protein (GFAP) and vimentin, and in most cases the two intermediate filament proteins co-localized. GFAP immunolabeling combined with immunohistochemistry against the mitotic marker 5-bromo-2'-deoxyuridine (BrdU) revealed an abundance of elongated BrdU-labeled nuclei closely apposed to, or localized within, GFAP-immunoreactive radial glia. The association of BrdU-labeled nuclei and GFAP-immunoreactive radial glial fibers was particularly pronounced 2 days after BrdU administration, when the migratory activity of the young cells is highest. When the new cells reach the granular layer, they start expressing the neuronal marker protein Hu C/D, but continue their close association with radial glial fibers. These results suggest the role of radial glia in the guidance of migrating adult-born neurons in the teleostean cerebellum. This function appears to be mediated both by somal translocation and by a glial-guided mode of locomotion.

Electric interactions through chirping behavior in the weakly electric fish, Apteronotus leptorhynchus.

The weakly electric fish Apteronotus leptorhynchus produces wave-like electric organ discharges distinguished by a high degree of regularity. Transient amplitude and frequency modulations ("chirps") can be evoked in males by stimulation with the electric field of a conspecific. During these interactions, the males examined in this study produced six types of chirps, including two novel ones. Stimulation of a test fish with a conspecific at various distances showed that two electrically interacting fish must be within 10 cm of each other to evoke chirping behavior in the neighboring fish. The chirp rate of all but one chirp type elicited by the neighboring fish was found to be negatively correlated with the absolute value of the frequency difference between the two interacting fish, but independent of the sign of this difference. Correlation analysis of the instantaneous rates of chirp occurrence revealed two modes of interactions characterized by reciprocal stimulation and reciprocal inhibition. Further analysis of the temporal relationship between the chirps generated by the two fish during electric interactions showed that the chirps generated by one individual follow the chirps of the other with a short latency of approximately 500-1,000 ms. We hypothesize that this "echo response" serves a communicatory function.