T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of α-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that α-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that α-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.     

Proline in the Osmoregulation of Brevibacterium lactofermentum

Osmoregulation in Brevibacterium lactofermentum was studied. Proline was accumulated up to approximately 35mg/g dry cell weight in the cells of a wild strain of the bacterium grown under osmotic stress. The osmotic tolerance of a proline auxotroph mutant obtained from the bacterium was lower than that in the wild strain. The activity of pyrroline-5-carboxylate reductase, one of the enzymes in the proline biosynthetic pathway, increased about 3-fold when the cells of B. lactofermentum were grown under osmotic stress. These data indicated that proline is important in osmoregulation in the bacterium.     

Effects of intra‐ and inter‐specific competition on fitness of sweetpotato weevil and West Indian sweetpotato weevil

The sweetpotato weevil Cylas formicarius (Fabricius) (Coleoptera: Brentidae) and West Indian sweetpotato weevil Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae) are major pests of sweet potato Ipomoea batatas (L.) Lam., in tropical and subtropical regions. Effects of intra‐ and inter‐specific competition on the number of progeny (fecundity), body weight and developmental time of C. formicarius and E. postfasciatus were examined in single‐ and mixed‐species treatments under laboratory conditions. Cylas formicarius tended to outcompete E. postfasciatus, whereas E. postfasciatus rather than C. formicarius tolerated higher conspecific densities. We discuss the implications of the results for pest management and resource partitioning of pestiferous weevils.     

Burr marigold (Bidens tripartita L.) roots directly and immediately scavenge rhizosphere methane with highly exuded hydrogen peroxide via a rhizosphere Fenton reaction

Plant and Soil - The major factors controlling the soil methane (CH4) concentration and CH4 emissions of various plant (mainly wetland) species were identified. Five plant species (Oryza sativa,...     

Soil carbon dynamics and aquatic metabolism of a wet–dry tropics wetland system

Freshwater wetlands are a key component of the global carbon cycle. Wet–dry tropics wetlands function as wet-season carbon sinks and dry-season carbon sources with low aquatic metabolism controlled by predictably seasonal, yet magnitude-variable flow regimes and inundation patterns. However, these dynamics have not been adequately quantified in Australia’s relatively unmodified wet–dry tropics freshwater wetlands. A baseline understanding is required before analysis of land-use or climate change impacts on these aquatic ecosystems can occur. This study characterises geomorphology and sedimentology within a seasonally connected wet–dry tropics freshwater wetland system at Kings Plains, Queensland, Australia, and quantifies soil carbon stocks and wet- and dry-season aquatic metabolism. Soil carbon stocks derived from loss-on-ignition on samples to 1 m depth were 51.5?±?7.8 kg C m^?2, higher than other wet–dry tropics wetlands globally, with potential for long-term retention at greater depths. Gross primary productivity of phytoplankton (GPP) and planktonic respiration (PR) measured through biological oxygen demand bottle experiments in the water column of sediment inundated under laboratory conditions show overall low GPP and PR in both wet- and dry-season samples (all wetland samples were heterotrophic with GPP/PR?<?1). Despite the short-term dominance of aquatic respiration processes leading to net release of carbon in the water column under these conditions, there is appreciable long-term storage of carbon in sediment in the Kings Plains wetlands. This demonstrates the importance of wet–dry-tropics wetland systems as hotspots of carbon sequestration, locally, regionally and globally, and consideration should be given to their conservation and management in this context.

     

Preparation of 8-Chloropurine Nucleosides Through the Reaction Between their C-8 Lithiated Species and p-Toluenesulfonyl Chloride

Abstract

Chlorination of purine nucleosides protected with tert-butyldimethylsilyl (TBDMS) group was examined by the reaction of the C-8 lithiated species, generated by LDA, with p-toluenesulfonyl chloride as an electrophile. This provides a new method for the preparation of 8-chloropurine nucleosides.     

Naemacyclus culmigenus,a newly reported potential pathogen to Miscanthus sinensis,new to Japan

Since the summer of 2010, a discomycete with erumpent apothecia associated with a leaf blight of Miscanthus leaves, were often collected. The morphological characteristics of the fungus suggested it was a member of the Helotiales rather than the Rhytismatales and this was supported by a phylogenetic analysis. Based on a morphological comparison with the type specimen of Naemacyclus culmigenus, currently known from Poaceae (Andropogon and Panicum), it was identified as N. culmigenus, new to Japan. The molecular phylogenetic analysis showed that the generic delimitation of Naemacyclus and related species requires clarification, as does their higher classification within the Leotiomycetes.     

Protein N-Myristoylation Plays a Critical Role in the Endoplasmic Reticulum Morphological Change Induced by Overexpression of Protein Lunapark,an Integral Membrane Protein of the Endoplasmic Reticulum

N-myristoylation of eukaryotic cellular proteins has been recognized as a modification that occurs mainly on cytoplasmic proteins. In this study, we examined the membrane localization, membrane integration, and intracellular localization of four recently identified human N-myristoylated proteins with predicted transmembrane domains. As a result, it was found that protein Lunapark, the human ortholog of yeast protein Lnp1p that has recently been found to be involved in network formation of the endoplasmic reticulum (ER), is an N-myristoylated polytopic integral membrane protein. Analysis of tumor necrosis factor-fusion proteins with each of the two putative transmembrane domains and their flanking regions of protein Lunapark revealed that transmembrane domain 1 and 2 functioned as type II signal anchor sequence and stop transfer sequence, respectively, and together generated a double-spanning integral membrane protein with an N-/C-terminal cytoplasmic orientation. Immunofluorescence staining of HEK293T cells transfected with a cDNA encoding protein Lunapark tagged with FLAG-tag at its C-terminus revealed that overexpressed protein Lunapark localized mainly to the peripheral ER and induced the formation of large polygonal tubular structures. Morphological changes in the ER induced by overexpressed protein Lunapark were significantly inhibited by the inhibition of protein N-myristoylation by means of replacing Gly2 with Ala. These results indicated that protein N-myristoylation plays a critical role in the ER morphological change induced by overexpression of protein Lunapark.