Disruption of the PDZ domain–binding motif of the dopamine transporter uniquely alters nanoscale distribution,dopamine homeostasis,and reward motivation

The dopamine (DA) transporter (DAT) is part of a presynaptic multiprotein network involving interactions with scaffold proteins via its C-terminal PDZ domain–binding sequence. Using a mouse model expressing DAT with mutated PDZ-binding sequence (DAT-AAA), we previously demonstrated the importance of this binding sequence for striatal expression of DAT. Here, we show by application of direct stochastic reconstruction microscopy not only that the striatal level of transporter is reduced in DAT-AAA mice but also that the nanoscale distribution of this transporter is altered with a higher propensity of DAT-AAA to localize to irregular nanodomains in dopaminergic terminals. In parallel, we observe mesostriatal DA adaptations and changes in DA-related behaviors distinct from those seen in other genetic DAT mouse models. DA levels in the striatum are reduced to ∼45% of that of WT, accompanied by elevated DA turnover. Nonetheless, fast-scan cyclic voltammetry recordings on striatal slices reveal a larger amplitude and prolonged clearance rate of evoked DA release in DAT-AAA mice compared with WT mice. Autoradiography and radioligand binding show reduced DA D2 receptor levels, whereas immunohistochemistry and autoradiography show unchanged DA D1 receptor levels. In behavioral experiments, we observe enhanced self-administration of liquid food under both a fixed ratio of one and progressive ratio schedule of reinforcement but a reduction compared with WT when using cocaine as reinforcer. In summary, our data demonstrate how disruption of PDZ domain interactions causes changes in DAT expression and its nanoscopic distribution that in turn alter DA clearance dynamics and related behaviors.     

Associations between Childbirth,Hospitalization and Disability Pension: A Cohort Study of Female Twins

Background

As the literature on long-term effects of childbirth on risk of morbidity or permanent work incapacity (DP) is limited, we aimed to study associations of childbirth with hospitalization and DP, adjusting for familial factors.

Methods

This cohort study included female twins, i.e. women with twin sister, born 1959–1990 in Sweden (n = 5 118). At least one in the twin pair had their first childbirth 1994–2009. Women were followed regarding all-cause and cause-specific (mental or musculoskeletal diagnoses) DP during year 2–5 after first delivery or equivalent. Associations between childbirth, hospitalization and DP were calculated as hazard ratios (HR) with 95% confidence intervals (CI).

Results

Women who did not give birth had markedly higher number of DP days/year compared to those giving birth. Hospitalization after first childbirth was associated with a higher HR of DP. Those hospitalized at least once after their first childbirth had a three-fold DP risk (HR: 3.2; 95% CI 1.1–9.6), DP due to mental diagnoses (HR: 3.2; 1.2–8.8), and of DP due to musculoskeletal diagnoses (HR: 6.1; 1.6–22.9). Lower HRs in the discordant twin pair analyses indicated that familial factors may influence the studied associations.

Conclusions

Women who did not give birth had a much higher risk for DP than those who did. Among those who gave birth, the risk for DP was markedly higher among those with a previous hospitalization, and especially in women with repeated hospitalizations. The results indicate a health selection into giving birth as well as the importance of morbidity for DP.     

Furanosesquiterpenes from the essential oil of myrrh

An examination of the essential oil of myrrh from Commiphora molmol has permitted the identification of 1(10)Z,4Z-furanodiene-6-one, 2-meth     

Acoustic signalling in female fish: factors influencing sound characteristics in croaking gouramis

The characteristics of sounds produced by fishes are influenced by several factors such as size. The current study analyses factors affecting structural properties of acoustic signals produced by female croaking gouramis Trichopsis vittata during agonistic interactions. Female sounds (although seldom analysed separately from male sounds) can equally be used to investigate factors affecting the sound characteristics in fish. Sound structure, dominant frequency and sound pressure levels (SPL) were determined and correlated to body size and the order in which sounds were emitted. Croaking sounds consisted of series of single-pulsed or double-pulsed bursts, each burst produced by one pectoral fin. Main energies were concentrated between 1.3 and 1.5 kHz. The dominant frequency decreased with size, as did the percentage of single-pulsed bursts within croaking sounds. The SPL and the number of bursts within a sound were independent of size but decreased significantly with the order of their production. Thus, acoustic signals produced at the beginning of agonistic interactions were louder and consisted of more bursts than subsequent ones. Our data indicate that body size affects the dominant frequency and structure of sounds. The increase in the percentage of double-pulsed bursts with size may be due to stronger pectoral muscles in larger fish. In contrast, ongoing fights apparently result in muscle fatigue and subsequently in a decline in the number of bursts and SPL. The factor ‘order of sound production’ points to an intra-individual variability of sounds and should be considered in future studies.     

A flash-photolysis study of the reactions of acaa 3-ttype cytochrome oxidase with dioxygen and carbon monoxide

The time course of absorbance changes following flash photolysis of the fully-reduced carboxycytochrome oxidase fromBacillus PS3 in the presence of O₂ has been followed at 445, 550, 605, and 830 nm, and the results have been compared with the corresponding changes in bovine cytochrome oxidase. The PS3 enzyme has a covalently bound cytochromec subunit and the fully-reduced species therefore accommodates five electrons instead of four as in the bovine enzyme. In the bovine enzyme, following CO dissociation, four phases were observed with time constants of about 10 s, 30 s, 100 s, and 1 ms at 445 nm. The initial, 10-s absorbance change at 445 nm is similar in the two enzymes. The subsequent phases involving hemea and Cu_A are not seen in the PS3 enzyme at 445 nm, because these redox centers are re-reduced by the covalently bound cytochromec, as indicated by absorbance changes at 550 nm. A reaction scheme consistent with the experimental observations is presented. In addition, internal electron-transfer reactions in the absence of O₂ were studied following flash-induced CO dissociation from the mixed-valence enzyme. Comparisons of the CO recombination rates in the mixed-valence and fully-reduced oxidases indicate that more electrons were transferred from hemea ₃ toa in PS3 oxidase compared to the bovine enzyme.     

Ecology, disturbance and restoration of coastal saltmarsh in Australia: a review

It is clear that saltmarshes are a unique and important component of the coastal biosphere of Australia. Their contribution ranges from stabilisation of fine sediments and providing an excellent protective buffer between land and sea, to their diverse blend of terrestrial and marine fauna. Further, saltmarsh plants are highly specialised and adapted to fill a harsh niche allowing them to act in roles that other vegetation types cannot. Saltmarsh habitats are recognised for their importance to migratory waders under the Ramsar convention, but it is becoming increasingly evident that they are also important to a variety of commercially valuable fish and native mammal species. Activities that are detrimental to saltmarshes continue and need to be addressed in order to conserve remaining saltmarsh areas. In general, urbanisation of the catchment has lead to filling of saltmarshes, tidal restriction, use by recreational vehicles, grazing, trampling and increased sedimentation and nutrient runnoff allowing colonisation and invasion of mangroves. These disturbances have a number of ecological consequences ranging from weed infestation to complete changes in the species composition and ecology. Reversing the disturbance is not always simple and can require extensive groundwork to be successful. Rehabilitation of existing saltmarsh areas has been a successful means to enhance this habitat. In general, it requires relatively little effort to remove weeds and fence off areas to regenerate naturally. Saltmarsh areas have been shown to respond well to this type of manipulation. Restoration and creation require substantial effort and planning to ensure a successful outcome. However, given the right environmental combinations of elevation, tide and salinity, saltmarsh will establish and grow. To speed the process transplantation of saltmarsh plants can be considered either from donor sites or plants propagated in green houses.     

Surrogate Habitats Demonstrate the Invasion Potential of the African Pugnacious Ant

Many ant species are highly invasive and are a significant component of disturbed ecosystems. They can have a major suppressive effect upon indigenous invertebrates, including other ants. Despite overwhelming circumstantial evidence for the ecological resourcefulness of many ants, there appears to be no experimental evidence illustrating the habitat breadth of a potentially invasive ant species. We demonstrate here that a particularly opportunistic and locally dominant ant Anoplolepis custodiens, which is a major indigenous African pest, overrides habitat structure to maintain its population level. We compared A. custodiens activity, morphology, foraging behaviour and ant species diversity in artificially established surrogate habitats (cover crops) in a vineyard containing an ample food resource in the form of the honeydew-producing mealybug Planococcus ficus. These cover crops were chosen so as to create highly altered habitats. The ant's ability to overcome these potentially suppressive habitat conditions hinged on its tight mutualism with the mealybug, and on its chasing away mealybug parasitoids. This ant species is predicted to be a latent invasive beyond Africa. It is unlikely to be impeded once it has established a foothold in a variety of novel habitats. It could locally invade to obtain food resources in a wide range of habitat types. Furthermore, in agricultural systems, cover crops are unlikely to control such an ant. Potential invasives such as this ant should be flagged as important quarantine suspects.     

Histone Deacetylase Inhibition Enhances Tissue Plasminogen Activator Release Capacity in Atherosclerotic Man

The expression of the tissue plasminogen activator (t-PA) gene appears to be under epigenetic control and can be affected by histone deacetylation inhibition. The study aimed to test if histone deacetalyase inhibitor treatment lead to increased t-PA release or reduced exhaustion in t-PA release in response to stimulation, as well as change in plasminogen activator inhibitor-1 (PAI-1) in subjects with coronary disease. In this clinical study, 16 post-myocardial infarction subjects, the perfused forearm model was used with isoprenaline provocation during 20 minutes, to stimulate local t-PA release. Each subject was measured twice on the same day (repeated stimuli sequences) as well as on two different occasions, without treatment and after four weeks of treatment with valproic acid (500 mg, twice daily). Net forearm release for t-PA in response to isoprenaline at minutes 1.5, 3, 6, 9, 12, 15 and 18 was measured, allowing assessment of cumulative t-PA release. There was a reduction in the exhaustion of cumulative t-PA release during repeated and prolonged stimulation with valproic acid treatment compared to non-treatment. Plasma PAI-1 antigen was decreased following treatment compared to non-treatment (18.4 ± 10.0 vs. 11.0 ± 7.1 nanograms/ml respectively, mean with 95% confidence interval). These findings demonstrate that histone deacetylation inhibition increases the capacity for endogenous t-PA release in subjects with vascular disease. Furthermore, the fibrinolytic balance is favored with suppressed PAI-1 levels. More studies are needed to establish the clinical relevance of these findings.

Trial registration

EU Clinical Trials Register 2012-004950-27     

A Novel Topology of Proline-rich Transmembrane Protein 2 (PRRT2): HINTS FOR AN INTRACELLULAR FUNCTION AT THE SYNAPSE*

Proline-rich transmembrane protein 2 (PRRT2) has been identified as the single causative gene for a group of paroxysmal syndromes of infancy, including epilepsy, paroxysmal movement disorders, and migraine. On the basis of topology predictions, PRRT2 has been assigned to the recently characterized family of Dispanins, whose members share the two-transmembrane domain topology with a large N terminus and short C terminus oriented toward the outside of the cell. Because PRRT2 plays a role at the synapse, it is important to confirm the exact orientation of its N and C termini with respect to the plasma membrane to get clues regarding its possible function. Using a combination of different experimental approaches, including live immunolabeling, immunogold electron microscopy, surface biotinylation and computational modeling, we demonstrate a novel topology for this protein. PRRT2 is a type II transmembrane protein in which only the second hydrophobic segment spans the plasma membrane, whereas the first one is associated with the internal surface of the membrane and forms a helix-loop-helix structure without crossing it. Most importantly, the large proline-rich N-terminal domain is not exposed to the extracellular space but is localized intracellularly, and only the short C terminus is extracellular (N_cyt/C_exo topology). Accordingly, we show that PRRT2 interacts with the Src homology 3 domain-bearing protein Intersectin 1, an intracellular protein involved in synaptic vesicle cycling. These findings will contribute to the clarification of the role of PRRT2 at the synapse and the understanding of pathogenic mechanisms on the basis of PRRT2-related neurological disorders.