Epigenetic alterations are critical for fear memory consolidation and synaptic plasticity in the lateral amygdala

Epigenetic mechanisms, including histone acetylation and DNA methylation, have been widely implicated in hippocampal-dependent learning paradigms. Here, we have examined the role of epigenetic alterations in amygdala-dependent auditory Pavlovian fear conditioning and associated synaptic plasticity in the lateral nucleus of the amygdala (LA) in the rat. Using Western blotting, we first show that auditory fear conditioning is associated with an increase in histone H3 acetylation and DNMT3A expression in the LA, and that training-related alterations in histone acetylation and DNMT3A expression in the LA are downstream of ERK/MAPK signaling. Next, we show that intra-LA infusion of the histone deacetylase (HDAC) inhibitor TSA increases H3 acetylation and enhances fear memory consolidation; that is, long-term memory (LTM) is enhanced, while short-term memory (STM) is unaffected. Conversely, intra-LA infusion of the DNA methyltransferase (DNMT) inhibitor 5-AZA impairs fear memory consolidation. Further, intra-LA infusion of 5-AZA was observed to impair training-related increases in H3 acetylation, and pre-treatment with TSA was observed to rescue the memory consolidation deficit induced by 5-AZA. In our final series of experiments, we show that bath application of either 5-AZA or TSA to amygdala slices results in significant impairment or enhancement, respectively, of long-term potentiation (LTP) at both thalamic and cortical inputs to the LA. Further, the deficit in LTP following treatment with 5-AZA was observed to be rescued at both inputs by co-application of TSA. Collectively, these findings provide strong support that histone acetylation and DNA methylation work in concert to regulate memory consolidation of auditory fear conditioning and associated synaptic plasticity in the LA.     

Formation of the zebrafish midbrain-hindbrain boundary constriction requires laminin-dependent basal constriction

The midbrain-hindbrain boundary (MHB) is a highly conserved fold in the vertebrate embryonic brain. We have termed the deepest point of this fold the MHB constriction (MHBC) and have begun to define the mechanisms by which it develops. In the zebrafish, the MHBC is formed soon after neural tube closure, concomitant with inflation of the brain ventricles. The MHBC is unusual, as it forms by bending the basal side of the neuroepithelium. At single cell resolution, we show that zebrafish MHBC formation involves two steps. The first is a shortening of MHB cells to approximately 75% of the length of surrounding cells. The second is basal constriction, and apical expansion, of a small group of cells that contribute to the MHBC. In the absence of inflated brain ventricles, basal constriction still occurs, indicating that the MHBC is not formed as a passive consequence of ventricle inflation. In laminin mutants, basal constriction does not occur, indicating an active role for the basement membrane in this process. Apical expansion also fails to occur in laminin mutants, suggesting that apical expansion may be dependent on basal constriction. This study demonstrates laminin-dependent basal constriction as a previously undescribed molecular mechanism for brain morphogenesis.     

Strong seasonal patterns of DOC release by a temperate seaweed community: Implications for the coastal ocean carbon cycle

Release of dissolved organic carbon (DOC) by seaweed underpins the microbial food web and is crucial for the coastal ocean carbon cycle. However, we know relatively little of seasonal DOC release patterns in temperate regions of the southern hemisphere. Strong seasonal changes in inorganic nitrogen availability, irradiance, and temperature regulate the growth of seaweeds on temperate reefs and influence DOC release. We seasonally surveyed and sampled seaweed at Coal Point, Tasmania, over 1 year. Dominant species with or without carbon dioxide (CO₂) concentrating mechanisms (CCMs) were collected for laboratory experiments to determine seasonal rates of DOC release. During spring and summer, substantial DOC release (10.06–33.54 μmol C · g DW⁻¹ · h⁻¹) was observed for all species, between 3 and 27 times greater than during autumn and winter. Our results suggest that inorganic carbon (C_i) uptake strategy does not regulate DOC release. Seasonal patterns of DOC release were likely a result of photosynthetic overflow during periods of high gross photosynthesis indicated by variations in tissue C:N ratios. For each season, we calculated a reef-scale net DOC release for seaweed at Coal Point of 7.84–12.9 g C · m⁻² · d⁻¹ in spring and summer, which was ~16 times greater than in autumn and winter (0.2–1.0 g C · m⁻² · d⁻¹). Phyllospora comosa, which dominated the biomass, contributed the most DOC to the coastal ocean, up to ~14 times more than Ecklonia radiata and the understory assemblage combined. Reef-scale DOC release was driven by seasonal changes in seaweed physiology rather than seaweed biomass.     

Intracellular calcium activity in split frog skin epithelium: Effect of cAMP

Summary Measurement of intracellular calcium activity (a _Ca ^c ) by ion-selective microelectrodes has previously been technically limited to relatively large cells (20 m). We now report results obtained with this technique in the small epithelial cells (10 m) of split frog skin using microelectrodes having an outer tip diameter of <0.2 m. The basolateral membrane potential was measured with Ca²⁺-selective microelectrodes (E _Ca ^sc ) and with reference micropipettes ( ^sc ) either sequentially or simultaneously in 15 successful experiments. Under baseline conditions,a _Ca ^c was measured to be 215±39nm (mean±se), in close agreement with the mean values estimated from published data obtained withNecturus proximal tubule. Stimulation of Na⁺ transport across six skins with 1mm serosal 8p-chlorophenylthio-3,5 cyclic AMP (CPTcAMP) increaseda _Ca ^c by a factor of 2.6±0.6. The increase ina _Ca ^c preceded the CPTcAMP-induced increase inI _sc. The results of the present study indicate that electrometric determination of intracellular calcium activity is now feasible in a much wider range of cell systems than heretofore possible. CPT cAMP elevates intracellular Ca²⁺ activity; this phenomenon is an early event, preceding the natriferic effect of CPTcAMP.     

A-kinase anchoring proteins interact with phosphodiesterases in T lymphocyte cell lines

The cAMP protein kinase A (PKA) pathway in T cells conveys an inhibitory signal to suppress inflammation. This study was performed to understand the mechanisms involved in cAMP-mediated signaling in T lymphocytes. A-kinase anchoring proteins (AKAPs) bind and target PKA to various subcellular locations. AKAPs also bind other signaling molecules such as cyclic nucleotide phosphodiesterases (PDEs) that hydrolyze cAMP in the cell. PDE4 and PDE7 have important roles in T cell activation. Based on this information, we hypothesized that AKAPs associate with PDEs in T lymphocytes. Immunoprecipitation of Jurkat cell lysates with Abs against both the regulatory subunit of PKA (RIIalpha) and specific AKAPs resulted in increased PDE activity associated with RIIalpha and AKAP95, AKAP149, and myeloid translocation gene (MTG) compared with control (IgG). Immunoprecipitation and pull-down analyses demonstrate that PDE4A binds to AKAP149, AKAP95, and MTG, but not AKAP79, whereas PDE7A was found to bind only MTG. Further analysis of MTG/PDE association illustrated that PDE4A and PDE7A bind residues 1-344 of MTG16b. Confocal analysis of HuT 78 cells stained with anti-PDE7A showed overlapping staining patterns with the Golgi marker GM130, suggesting that PDE7A is located in the Golgi. The staining pattern of PDE7A also showed similarity to the staining pattern of MTG, supporting the immunoprecipitation data and suggesting that MTG may interact with PDE7A in the Golgi. In summary, these data suggest that AKAPs interact with both PKA and PDE in T lymphocytes and thus are a key component of the signaling complex regulating T cell activation.     

Galectins in teleost fish: Zebrafish (Danio rerio) as a model species to address their biological roles in development and innate immunity

Cell surface glycans, such as glycocoproteins and glycolipids, encode information that modulates interactions between cells, or between cells and the extracellular matrix, by specifically regulating the binding to cell surface-associated or soluble carbohydrate-binding receptors, such as lectins. Rapid modifications of exposed carbohydrate moieties by glycosidases and glycosyltransferases, and the equally dynamic patterns of expression of their receptors during early development, suggest that both play important roles during embryogenesis. Among a variety of biological roles, galectins have been proposed to mediate developmental processes, such as embryo implantation and myogenesis. However, the high functional redundancy of the galectin repertoire in mammals has hindered the rigorous characterization of their specific roles by gene knockout approaches in murine models. In recent years, the use of teleost fish as alternative models for addressing developmental questions in mammals has expanded dramatically, and we propose their use for the elucidation of biological roles of galectins in embryogenesis and innate immunity. All three major galectin types, proto, chimera, and tandem-repeat, are present in teleost fish, and phylogenetic topologies confirm the expected clustering with their mammalian orthologues. As a model organism, the zebrafish (Danio rerio) may help to overcome limitations imposed by the murine models because it offers substantial advantages: external fertilization, transparent embryos that develop rapidly in vitro, a diverse toolbox of established methods to manipulate early gene expression, a growing collection of mutations that affect early embryonic development, availability of cell lines, and most importantly, an apparently less diversified galectin repertoire. Published in 2004.     

The red-vented bulbul (<Emphasis Type="Italic">Pycnonotus cafer</Emphasis>): serious pest or understudied invader?

Recently, debate has flourished about inadequacies in the simplistic “worst invasive species” approach and its global scale. Here we investigate the status of the red-vented bulbul (Pycnonotus cafer), an Asian passerine bird. This species has been introduced widely across Pacific islands and is commonly blamed for its impacts on agriculture and biodiversity via dispersal of invasive plant seeds and competition with native fauna. This case study evaluates all available data on the impacts and management of this invasive species and identifies priorities for future research. We reviewed the scientific literature and information from three databases (ABBA, GAVIA, eBird) and highlight that the attention paid to this species by scientists and managers varied considerably between islands and contexts and was globally lower than the attention paid to other species on the IUCN-ISSG list. The red-vented bulbul has now established on 37 islands and in seven continental locations outside its native range. We show that three categories of effects are associated with this species: plant damage, seed dispersal and disturbance of fauna. We compiled lists of 110 plant species consumed, 33 plant species dispersed, and 15 species of bird that this bulbul interacts with. However, these lists were mainly made of opportunistic observations rather than specific assessments. Research outputs that focus on better ways to prevent or quantify the impacts of the red-vented bulbul remain scarce. We found very few references exploring potential positive impacts of this species, and only two examples of management actions undertaken against it. The latter are required to inform management actions, especially on sensitive tropical islands where invasions and dispersal of the red-vented bulbul are ongoing. Our analysis of the literature found no clear support for considering this species to be one of the “world’s worst” invasive alien species.