Plasticity in the hippocampal formation of shorebirds during the wintering period: Stereological analysis of parvalbumin neurons in Actitis macularius.

The number of parvalbumin neurons can be modified by social, multisensory, and cognitive stimuli in both mammals and birds, but nothing is known about their plasticity in long-distance migratory shorebirds. Here, in the spotted sandpiper (Actitis macularius), we investigated the plasticity of parvalbumin neurons of two brain areas during this species' wintering period at a lower latitude. We compared individuals in a nonmigratory rest period (November-January) and premigration (May-July) period. We used parvalbumin as a marker for counting a subpopulation of inhibitory neurons in the hippocampal formation (HF), with the magnocellular nucleus of the tectal isthmus (IMC) as a control area. Because the HF is involved in learning and memory and social interaction and the IMC is essential for control of head, neck, and eye movements, we hypothesized that parvalbumin neurons would increase in the HF and remain unchanged in the IMC. We used an optical fractionator to estimate cell numbers. Compared with the nonmigratory rest birds, parvalbumin neuron count estimates in the premigration birds increased significantly in the HF but remained unchanged in IMC. We suggest that the greater number of parvalbuminergic neurons in the HF of A. macularius in the premigration period represents adaptive circuitry changes involved in the migration back to reproductive niches in the northern hemisphere.

Shorebirds' Longer Migratory Distances Are Associated With Larger ADCYAP1 Microsatellites and Greater Morphological Complexity of Hippocampal Astrocytes.

For the epic journey of autumn migration, long-distance migratory birds use innate and learned information and follow strict schedules imposed by genetic and epigenetic mechanisms, the details of which remain largely unknown. In addition, bird migration requires integrated action of different multisensory systems for learning and memory, and the hippocampus appears to be the integration center for this task. In previous studies we found that contrasting long-distance migratory flights differentially affected the morphological complexity of two types of hippocampus astrocytes. Recently, a significant association was found between the latitude of the reproductive site and the size of the ADCYAP1 allele in long distance migratory birds. We tested for correlations between astrocyte morphological complexity, migratory distances, and size of the ADCYAP1 allele in three long-distance migrant species of shorebird and one non-migrant. Significant differences among species were found in the number and morphological complexity of the astrocytes, as well as in the size of the microsatellites of the ADCYAP1 gene. We found significant associations between the size of the ADCYAP1 microsatellites, the migratory distances, and the degree of morphological complexity of the astrocytes. We suggest that associations between astrocyte number and morphological complexity, ADCYAP1 microsatellite size, and migratory behavior may be part of the adaptive response to the migratory process of shorebirds.

Changes in hippocampal astrocyte morphology of Ruddy turnstone (Arenaria interpres) during the wintering period at the mangroves of Amazon River estuary.

Astrocytes are essential for lipid neuronal metabolism in long-distance uninterrupted migratory flights, when glucose is not available as the main source of energy. We previously demonstrated in Calidris pusilla that after uninterrupted 5 days transatlantic flight, astrocytes shrink and reduce its number in the hippocampal formation. Here we shifted our attention to the wintering period and tested the hypothesis that hippocampal astrocyte morphology of A interpres will change as the wintering period progresses towards the premigration window. To that end we used Arenaria interpres, which also crosses the Atlantic Ocean and reaches the mangroves of the Amazon River estuary for wintering. Birds were captured in September/October (closer to the arrival in the coast of Bragança, Para, Brazil for wintering) and in April/May (closer to the departure towards the breeding sites) and had their brains processed for selective GFAP-astrocyte immunolabeling. Three-dimensional reconstructions of the immunostained astrocytes were performed and morphological classification was done based on hierarchical cluster and discriminant analysis of multimodal morphometric features. We found two morphological phenotypes of astrocytes in the newcomers which differentially increased its morphological complexities as wintering period progresses towards the pre-migration window. Taken together, our findings demonstrate that the long-distance non-stop flight and wintering period differentially affected the two astrocytes morphotypes, suggesting distinct physiological roles for these cells. We suggest that morphological changes during the wintering period, may be part of the adaptive plasticity of the local hippocampal circuits of A. interpres in preparation for the long journey back to their breeding sites in the north hemisphere.

Differential Microglial Morphological Response, TNFα, and Viral Load in Sedentary-like and Active Murine Models After Systemic Non-neurotropic Dengue Virus Infection.

Peripheral inflammatory stimuli increase proinflammatory cytokines in the bloodstream and central nervous system and activate microglial cells. Here we tested the hypothesis that contrasting environments mimicking sedentary and active lives would be associated with differential microglial morphological responses, inflammatory cytokines concentration, and virus load in the peripheral blood. For this, mice were maintained either in standard (standard environment) or enriched cages (enriched environment) and then subjected to a single (DENV1) serotype infection. Blood samples from infected animals showed higher viral loads and higher tumor necrosis factor-α (TNFα) mRNA concentrations than control subjects. Using an unbiased stereological sampling approach, we selected 544 microglia from lateral septum for microscopic 3D reconstruction. Morphological complexity contributed most to cluster formation. Infected groups exhibited significant increase in the microglia morphological complexity and number, despite the absence of dengue virus antigens in the brain. Two microglial phenotypes (type I with lower and type II with higher morphological complexity) were found in both infected and control groups. However, microglia from infected mice maintained in enriched environment showed only one morphological phenotype. Two-way ANOVA revealed that environmental changes and infection influenced type-I and II microglial morphologies and number. Environmental enrichment and infection interactions may contribute to microglial morphological change to a point that type-I and II morphological phenotypes could no longer be distinguished in infected mice from enriched environment. Significant linear correlation was found between morphological complexity and TNFα peripheral blood. Our findings demonstrated that sedentary-like and active murine models exhibited differential microglial responses and peripheral inflammation to systemic non-neurotropic infections with DENV1 virus.