Influence of fear conditioning on elicited ponto-geniculo-occipital waves and rapid eye movement sleep

The amygdala plays a central role in fear conditioning, a model of anticipatory anxiety. It has massive projections to brainstem regions involved in rapid eye movement sleep (REM) and ponto-geniculo-occipital (PGO) wave generation. PGO waves occur spontaneously in REM or in response to stimuli. Electrical stimulation of the central nucleus of the amygdala enhances spontaneous PGO wave activity during REM and the amplitude of both the acoustic startle response and the elicited PGO wave (PGOE), a neural marker of alerting. This study examined the effects of fear conditioning on REM and on PGOE. On conditioning days, the number of REM episodes, the average REM duration and the REM percentage were decreased while REM latency was increased. The presentation of auditory stimuli in the presence of a light conditioned stimulus produced PGOE of greater amplitudes. The results suggest that fear, most likely involving the amygdala, can influence REM and brainstem alerting mechanisms.     

Effects of Brassica oleracea waxblooms on predation and attachment by Hippodamia convergens

Four mutations that reduce waxbloom in Brassica oleracea L. were examined for their effects on predation, mobility, and adhesion to the plant surface by the general predator Hippodamia convergens (Guérin-Menéville) (Coleoptera: Coccinellidae). The mutations reduce waxbloom to different degrees, but all produce a glossy phenotype. Plants tested were inbred lines, near isogenic lines, or segregating F₂ populations, depending on the mutation. In an experiment on caged leaves, predation of Plutella xylostella L. larvae by H. convergens adult females was significantly greater on glossy types as compared with normal-wax or wild-type counterparts. Although the trend was the same for each mutation, individual comparisons between glossy and normal-wax lines or segregants were only significant for two of them, those producing mutant alleles gl_a and gl_d. Individual H. convergens were observed to spend more time walking on leaf edges and less time walking on leaf surfaces of normal-wax plants than glossy plants. Hippodamia convergens also obtained better adhesion to the surfaces of glossy plants than to normal-wax plants when tested using a centrifugal device. Two of the mutations produced similarly strong effects on predation, behaviour, and adhesion by H. convergens. These two are the same previously determined by us to provide the strongest similar effects on another generalist predator, Chrysoperla plorabunda (Fitch). The results indicate that waxbloom variation in nature could affect herbivore populations through its effects on generalist predators.     

Predation, behavior, and attachment by Chrysoperla plorabunda larvae on Brassica oleracea with different surface waxblooms

Predation by Chrysoperla plorabunda (Fitch) (Chrysopidae) first instars on Plutella xylostella (L.) (Plutellidae) neonates was measured on nine Brassica oleracea L. types with different surface wax crystal morphology. Forty-eight-hour survival of P. xylostella caged on leaves was significantly reduced by C. plorabunda on plants expressing glossy mutations that reduce surface waxbloom, but not on plants with normal waxbloom. During 5-min observations, C. plorabunda spent more time walking and less time scrambling (moving the legs with no locomotion) on glossy types than on the normal-wax types. Adhesive force produced by C. plorabunda on plant surfaces was 20 to 200-fold greater on glossy types than on normal-wax types. All the glossy types provided an advantage to C. plorabunda over normal-wax types by increasing the force of attachment to the leaf suface by the larvae, which in turn increased time allocated to walking, leading to greater predation of P. xylostella larvae. Among glossy and normal wax types together, attachment force and time allocated to walking were significantly correlated with predation by C. plorabunda. Within glossy or normal-wax types, however, these correlations were not significant. Neither time allocated to walking, nor attachment to the leaf surface was a predictor of predation by C. plorabunda within glossy or normal-wax types. Although diverse mechanisms therefore contribute to differences in predation, the results show that reduction in waxbloom can substantially affect the mobility and effectiveness of this generalist predator.     

Extremity hyperinsulinemia stimulates muscle protein synthesis in severely injured patients

Insulin has a well-recognized anabolic effect on muscle protein, yet critically ill, severely injured patients are often considered "resistant" to the action of insulin. The purpose of this study was to assess the in vivo effects of hyperinsulinemia on human skeletal muscle in severely injured patients. To accomplish this goal, 14 patients with burns encompassing >40% of their body surface area underwent metabolic evaluation utilizing isotopic dilution of phenylalanine, femoral artery and vein blood sampling, and sequential muscle biopsies of the leg. After baseline metabolic measurements were taken, insulin was infused into the femoral artery at 0.45 mIU.min(-1).100 ml leg volume(-1) to create a local hyperinsulinemia but with minimal systemic perturbations. Insulin administration increased femoral venous concentration of insulin (P < 0.01) but with only a 4% (insignificant) decrease in the arterial glucose concentration and a 7% (insignificant) decrease in the arterial concentration of phenylalanine. Extremity hyperinsulinemia significantly increased leg blood flow (P < 0.05) and the rate of muscle protein synthesis (P < 0.05). Neither the rate of muscle protein breakdown nor the rate of transmembrane transport of phenylalanine was significantly altered with extremity hyperinsulinemia. In conclusion, this study demonstrates that insulin directly stimulates muscle protein synthesis in severely injured patients.     

In vitro and in vivo effects of GABA, muscimol, and bicuculline on lamprey GnRH concentration in the brain of the sea lamprey (Petromyzon marinus)

gamma-Aminobutyric acid (GABA) is a neurotransmitter with a demonstrated neuroregulatory role in reproduction in most representative species of vertebrate classes via the hypothalamus. The role of GABA on the hypothalamus-pituitary axis in lampreys has not been fully elucidated. Recent immunocytochemical and in situ hybridization studies suggest that there may be a neuroregulatory role of GABA on the gonadotropin-releasing hormone (GnRH) system in lampreys. To assess possible GABA-GnRH interactions, the effects of GABA and its analogs on lamprey GnRH in vitro and in vivo were studied in adult female sea lampreys (Petromyzon marinus). In vitro perfusion of GABA and its analogs at increasing concentrations (0.1-100 microM) was performed over a 3-h time course. There was a substantial increase of GnRH-I and GnRH-III following treatment of muscimol at 100 microM. In in vivo studies, GABA or muscimol injected at 200 microg/kg significantly increased lamprey GnRH concentration in the brain 0.5 h after treatment compared to controls in female sea lampreys. No significant change in lamprey GnRH-I or GnRH-III was observed following treatment with bicuculline. These data provide novel physiological data supporting the hypothesis that GABA may influence GnRH in the brain of sea lamprey.     

Synaptotagmin VII restricts fusion pore expansion during lysosomal exocytosis

Synaptotagmin is considered a calcium-dependent trigger for regulated exocytosis. We examined the role of synaptotagmin VII (Syt VII) in the calcium-dependent exocytosis of individual lysosomes in wild-type (WT) and Syt VII knockout (KO) mouse embryonic fibroblasts (MEFs) using total internal reflection fluorescence microscopy. In WT MEFs, most lysosomes only partially released their contents, their membrane proteins did not diffuse into the plasma membrane, and inner diameters of their fusion pores were smaller than 30 nm. In Syt VII KO MEFs, not only was lysosomal exocytosis triggered by calcium, but all of these restrictions on fusion were also removed. These observations indicate that Syt VII does not function as the calcium-dependent trigger for lysosomal exocytosis. Instead, it restricts the kinetics and extent of calcium-dependent lysosomal fusion.     

Tracking metastatic tumor cell extravasation with quantum dot nanocrystals and fluorescence emission-scanning microscopy

Metastasis is an impediment to the development of effective cancer therapies. Our understanding of metastasis is limited by our inability to follow this process in vivo. Fluorescence microscopy offers the potential to follow cells at high resolution in living animals. Semiconductor nanocrystals, quantum dots (QDs), offer considerable advantages over organic fluorophores for this purpose. We used QDs and emission spectrum scanning multiphoton microscopy to develop a means to study extravasation in vivo. Although QD labeling shows no deleterious effects on cultured cells, concern over their potential toxicity in vivo has caused resistance toward their application to such studies. To test if effects of QD labeling emerge in vivo, tumor cells labeled with QDs were intravenously injected into mice and followed as they extravasated into lung tissue. The behavior of QD-labeled tumor cells in vivo was indistinguishable from that of unlabeled cells. QDs and spectral imaging allowed the simultaneous identification of five different populations of cells using multiphoton laser excitation. Besides establishing the safety of QDs for in vivo studies, our approach permits the study of multicellular interactions in vivo.     

UCS proteins: managing the myosin motor

Recent studies indicate that myosin molecular motors interact inside cells with proteins containing a conserved 'UCS' domain. This appears to ensure proper folding of myosin heads so that they can perform their ATP-dependent actin-based motor functions.     

Specific interaction of protein tyrosine phosphatase-MEG2 with phosphatidylserine

Protein tyrosine phosphatase (PTP)-MEG2 is an intracellular tyrosine phosphatase that contains a Sec14 homology domain. We have purified the full-length and truncated forms of the enzyme from recombinant adenovirus-infected human 293 cells. By using lipid-membrane overlay and liposome binding assays, we demonstrated that PTP-MEG2 specifically binds phosphatidylserine among over 20 lipid compounds tested. The binding is mediated by its N-terminal Sec14 domain. In intact cells, the Sec14 domain is responsible for localization of PTP-MEG2 to the perinuclear region, and uploading of PS into the cell membrane causes translocation of PTP-MEG2 to the plasma membrane. Phosphatidylserine is a relatively abundant cell membrane phospholipid non-symmetrically distributed in the outer layer and inner layer of cell membranes. It has recently been defined as an important ligand for clearance of apoptotic cells. By specifically binding phosphatidylserine, PTP-MEG2 may play an important role in regulating signaling processes associated with phagocytosis of apoptotic cells.