Rapid lesioning of large numbers of identified vertebrate neurons: applications in zebrafish.

Establishing a causal role between the activity of specific individual nerve cells and the behaviors they produce (or the neural computations they execute) is made difficult in vertebrate animals because of the large numbers of neurons involved. Traditional techniques for establishing causal roles, such as tract cutting and electrolytic lesions, are limited because they produce damage that affects a variety of different cell types, invariably intermingled, and often of uncertain identity. We propose here an alternative lesioning technique in which large numbers of neurons are lesioned, but the lesioned neurons are specifically identified by fluorescent labeling. We use the locomotor control system of the larval zebrafish to illustrate this approach. In this example, the technique involves injection of fluorescent dextrans into far-rostral spinal cord to label descending nerve fibers. Such injections appear to interrupt the descending nerve fibers, and therefore their accompanying locomotor control signals. This protocol is shown to produce significant behavioral deficits. Because the CNS of the larval zebrafish is transparent, the entire population of lesioned cells can be imaged in vivo and reconstructed using confocal microscopy. This large-scale lesioning technique is important, even in this relatively 'simple' vertebrate animal, because the ablation of smaller numbers of neurons, using more precise laser-ablation techniques, often fails to produce observable behavioral deficits. While this technique is most readily applied in simpler and transparent vertebrate animals, the approach is general in nature and might, in principle, be applied to any vertebrate nerve tract.

Performance more than working memory disrupted by acute systemic inflammation in rats in appetitive tasks.

Evidence from molecular biology, epidemiology, behavioral pharmacology, and clinical science support the conclusion that brain inflammation contributes to the pathogenesis of cognitive symptoms in Alzheimer's disease (AD) and other neuropsychological disorders. Three different tests were conducted to determine whether the acute inflammatory response induced by systemic lipopolysaccharide (LPS) treatment is accompanied by a selective disruption of working memory functioning in rats. Doses of LPS sufficient to induce a thermoregulatory response were administered intraperitoneally and their effects on behavioral measures of symbolic working memory, spatial learning, and spatial memory consolidation, were assessed. LPS-induced immune activation was found not to significantly affect memory processes in any of the behavioral tests used. However, LPS-induced immune activation caused performance deficits consistent with a disruptive effect of LPS on motivation and arousal. These results suggest that sickness behavior induced by immune stimulation is not necessarily accompanied by selective impairment in memory processes. The importance of distinguishing cognitive disruption from performance impairment in interpreting the behavioral effects of inflammatory mediators is discussed.

Ibuprofen effects on Alzheimer pathology and open field activity in APPsw transgenic mice.

We previously showed the non-steroidal anti-inflammatory drug (NSAID) ibuprofen suppresses inflammation and amyloid in the APPsw (Tg2576) Tg2576 transgenic mouse. The mechanism for these effects and the impact on behavior are unknown. We now show ibuprofen's effects were not mediated by alterations in amyloid precursor protein (APP) expression or oxidative damage (carbonyls). Six months ibuprofen treatment in Tg+ females caused a decrease in open field behavior (p < 0.05), restoring values similar to Tg- mice. Reduced caspase activation per plaque provided further evidence for a neuroprotective action of ibuprofen. The impact of a shorter 3 month duration ibuprofen trial, beginning at a later age (from 14 to 17 months), was also investigated. Repeated measures ANOVA of Abeta levels (soluble and insoluble) demonstrated a significant ibuprofen treatment effect (p < 0.05). Post-hoc analysis showed that ibuprofen-dependent reductions of both soluble Abeta and Abeta42 were most marked in entorhinal cortex (p < 0.05). Although interleukin-1beta and insoluble Abeta were more effectively reduced with longer treatment, the magnitude of the effect on soluble Abeta was not dependent on treatment duration.

Inflammatory pathogenesis in Alzheimer's disease: biological mechanisms and cognitive sequeli.

Experimental evidence from molecular biology, biochemistry, epidemiology and behavioral research support the conclusion that brain inflammation contributes to the pathogenesis of Alzheimer's disease and other types of human dementias. Aspects of neuroimmunology relating to the pathogenesis of Alzheimer's disease are briefly reviewed. The effects of brain inflammation, mediated through cytokines and other secretory products of activated glial cells, on neurotransmission (specifically, nitric oxide, glutamate, and acetylcholine), amyloidogenesis, proteolysis, and oxidative stress are discussed within the context of the pathogenesis of learning and memory dysfunction in Alzheimer's disease. Alzheimer's disease is proposed to be an etiologically heterogeneous syndrome with the common elements of amyloid deposition and inflammatory neuronal damage.

Reversible impairment of long-term potentiation in transgenic Cu/Zn-SOD mice.

Copper/zinc superoxide dismutase (CuZn-SOD) is a key enzyme in the metabolism of oxygen free radicals. The gene encoding CuZn-SOD resides on human chromosome 21 and is overexpressed in Down syndrome (DS) patients. Overexpression of CuZn-SOD in transgenic (Tg) mice and cultured cells creates chronic oxidative stress leading to enhanced susceptibility to degeneration and apoptotic cell death. We have now found that three lines of Tg-CuZn-SOD mice, one of which also overexpresses S100beta, a glial calcium binding protein, are deficient in spatial memory. Furthermore, hippocampal slices taken from these mice have an apparently normal synaptic physiology, but are impaired in the ability to express long-term potentiation (LTP). This effect on hippocampal LTP was abrogated by treatment of slices with the H2O2 scavenger catalase or the antioxidant N-t-butyl-phenylnitrone (BPN). It is proposed that elevated CuZnSOD causes an increase in tetanic stimulation-evoked formation of H2O2 which leads to diminished LTP and cognitive deficits in these mice.

The relationships among saccharin consumption, oral ethanol, and i.v. cocaine self-administration.

The purpose of the present experiment was to replicate previously reported observations of a relationship between saccharin consumption and oral ethanol self-administration in rats using operant measures (2,8) and to determine whether saccharin intake was related to the rate of acquisition of IV cocaine self-administration. Groups of Wistar rats selected for high and low saccharin (0.1% wt/vol) intake were tested for rate of acquisition of IV cocaine (0.2 mg/kg/infusion) self-administration using an autoshaping procedure. They were subsequently tested for self-administration of oral ethanol (8% wt/vol) under ascending fixed-ratio (FR) schedules (FR 1, 2, 4, and 8). Finally, ethanol deliveries were compared under food-deprivation and food-satiation conditions under an FR 8 schedule. Saccharin intake was redetermined after each phase of the experiment. No significant differences between high and low saccharin groups were found in rate of acquisition of IV cocaine self-administration, and there was not a significant correlation between saccharin and cocaine consumption. However, the high saccharin group drank significantly more ethanol than the low saccharin group during the FR 8 food satiation component. A significant correlation between saccharin and ethanol consumption was also found. For high and low saccharin groups, responding for ethanol increased proportionally with increases in FR such that consumption of ethanol remained relatively constant as FR increased. Ethanol consumption was significantly increased under food deprivation relative to food satiation conditions for both saccharin groups. A significant correlation between ethanol consumption and cocaine consumption was also found. Significant increases in saccharin consumption across successive saccharin consumption tests were found for both groups, although relative intake for the high and low saccharin groups remained stable throughout the experiment. These results indicate that higher ethanol intake is predicted by higher saccharin intake, but saccharin intake did not predict the rate of acquisition of IV cocaine self-administration.