Leghemoglobin: Different roles for different components?

The ratio of leghemoglobin c content to leghemoglobin a content in soybean (Glycine max) root nodules drops in the early stages of plant growth and becomes constant during flowering and fruiting. These data bring into question the assumption that leghemoglobin components have a single biochemical role in nitrogen fixation.     

What couples glycolysis to mitochondrial signal generation in glucose-stimulated insulin secretion?

Pancreatic islet beta-cells are poised to generate metabolic messengers in the mitochondria that link glucose metabolism to insulin exocytosis. This is accomplished through the tight coupling of glycolysis to mitochondrial activation. The messenger molecules ATP and glutamate are produced after the metabolism of glycolysis-derived pyruvate in the mitochondria. The entry of monocarboxylates such as pyruvate into the beta cell is limited, explaining why overexpression of monocarboxylate transporters unravels pyruvate-stimulated insulin secretion. NADH generated by glycolysis is efficiently reoxidized by highly active mitochondrial shuttles rather than by lactate dehydrogenase. Overexpression of this enzyme does not alter glucose-stimulated insulin secretion, suggesting that NADH availability restricts the conversion of pyruvate to lactate in the beta cell. These metabolic features permit the fuel function of glucose to be extended to the generation of signaling molecules, which increases cytosolic Ca2+ and promotes insulin exocytosis.     

Putative roles for phospholipase Cη enzymes in neuronal Ca2+ signal modulation

The most recently identified PLC (phospholipase C) enzymes belong to the PLCη family. Their unique Ca2+-sensitivity and their specific appearance in neurons have attracted great attention since their discovery; however, their physiological role(s) in neurons are still yet to be established. PLCη enzymes are expressed in the neocortex, hippocampus and cerebellum. PLCη2 is also expressed at high levels in pituitary gland, pineal gland and in the retina. Driven by the specific localization of PLCη enzymes in different brain areas, in the present paper, we discuss the roles that they may play in neural processes, including differentiation, memory formation, circadian rhythm regulation, neurotransmitter/hormone release and the pathogenesis of neurodegenerative disorders associated with aberrant Ca2+ signalling, such as Alzheimer's disease.     

Pollination and stigma wounding: same response, different signal?

In Petunia hybrida flowers, both pollination and stigma woundinginduced a transient Increase in ethylene production and hastenedcorolla senescence. Ethylene production by different flowerparts was measured in situ using laser photoacoustic (LPA) spectroscopy.In pollinated flowers, ethylene was exclusively produced bythe stigma/style region whereas wounding of the stigma Inducedethylene production both by the stigma/style region and by theremaining flower parts. In aminoethoxyvinylglycine (AVG)-treatedflowers, subsequent treatment of the unwounded stigma with 1-aminocyclopropane-1-carboxylicacid (ACC) induced ethylene production exclusively by the stigma/styleregion whereas treatment of a previously wounded stigma withACC induced a simultaneous increase in ethylene production bythe stigma/style region and the remaining flower parts. Theseresults suggest that following stigma wounding, either ACC orethylene is involved in inter-organ communication. Followingpollination, the signal is apparently not directly related toethylene. In vivo ACC oxidase activity of most flower parts, includingthe gynoecium, was higher in light than in dark. Light or darkdid not influence the relative contributions of stigma/styleand remaining flower parts to the total pollination, woundingor ACC-induced ethylene production, indicating that ACC is nottranslocated. Both in excised styles and intact flowers, radiolabelledACC and its analogue -aminoisobutyric acid (AIB), applied eitherto an intact or wounded stigma, were largely immobile confirmingthat ACC is not likely to play a role in inter-organ signalling. The results collectively suggest that following stigma wounding,translocation of ethylene may be the signal responsible forinitiation of corolla senescence; following pollination thesignal is not directly related to ethylene. Key words: 1-Aminocyclopropane-1-carboxylic acid (ACC), ethylene, flower senescence, Petunia hybrida, pollination, stigma wounding     

Are germination cues for soil‐stored seed banks different in structurally different fire‐prone communities?

Many plant species are dependent on soil‐stored seeds for their persistence in fire‐prone systems. Seed germination is often stimulated by fire‐related cues including heat and smoke, but the way these cues promote germination may differ between structurally distinct plant communities with historically different fire regimes. In this study, we examined the effects of heat, smoke and their interactions on the germination of soil‐stored seeds from shrubby woodlands and herbaceous forests in south‐east Australia. The effect of these treatments on species richness, diversity and composition, and species richness and density of germinants within life‐forms (grass, forb and shrub) were assessed. Soils from each community were subjected to low heat (40°C), low heat with smoke, smoke, high heat (80°C), high heat with smoke and untreated (control) before being placed in a glasshouse, where the germinants were identified and counted. Greater species richness was stimulated by high heat treatments and smoke in both communities, a trend driven by shrubs and forbs, rather than grasses. Greater species diversity was stimulated by high heat with smoke in both communities. Greater densities of grass germinants were stimulated by all treatments, except low heat, in both communities. For forbs and shrubs, the effect of treatment depended on community. Compared to the control, low heat with smoke (forbs) and both low heat and low heat with smoke (shrubs) increased densities in the woodland but not in the forest. There were unique species compositions, different from the control, in all treatments in the forest but not in the woodland, where composition in low heat was not different from the control. These results indicate the importance of high soil temperatures and smoke in both communities. In the absence of wildfires, recurring prescribed burns that heat the soil to low temperatures are likely to reduce plant richness, diversity, and density resulting in a change in understorey species composition and structure.     

A second signal for autophagic cell death?

Dictyostelium cells in monolayers in vitro lend themselves well to a study of autophagic cell death (ACD). There is no apoptosis machinery in the protist Dictyostelium, no caspase nor Bcl-2 family members (except a paracaspase whose inactivation does not alter cell death), thus there is no apoptosis that could interfere with, or substitute for, non-apoptotic cell death. Also, Dictyostelium, a eukaryote, has a haploid genome, which facilitates random insertional mutagenesis.     

Is extracellular calcium required for insulin action?

Isolated hepatocytes and isolated adipocytes incubated in the absence of added calcium ions respond to insulin with a decrease in tissue cyclic AMP levels, and an increase in low Km phosphodiesterase activity. Isolated hepatocytes also showed a diminution of glucagon stimulated glucose output in response to insulin, while adipocytes responded with increased rates of glucose oxidation, lipid synthesis and decreased glycerol output. These responses to insulin are the same as those seen when the cells are incubated in buffers containing physiological concentrations of calcium ions. When extracellular concentrations of calcium ions were made extremely low by using either gelatine or albumin which had been pretreated to remove calcium, and/or the incubation buffers contained EGTA, both the hepatocytes and adipocytes continued to respond to insulin. These results suggest that extracellular calcium ions are not required for insulin action.     

Salmonella entry into mammalian cells: different yet converging signal transduction pathways?

Salmonella bacteria have evolved means to subvert host cell signal transduction pathways to induce their uptake. Recently, progress has been made towards defining those pathways. Although it is clear that Salmonella evoke different signalling pathways in different cell lines, it is possible that these responses may be triggered by a common mechanism and that the diverse pathways may converge downstream in common effector molecules.     

Monoubiquitination: the signal for p53 nuclear export?

The ubiquitin-proteasome pathway has become an increasingly important regulatory mechanism for protein function. Countless proteins are degraded by the addition of polymeric ubiquitin chains, but more recently, monoubiquitination has emerged as a mechanism for regulatory functions other than proteasomal degradation. Monoubiquitination acts as a signal in nuclear export for the tumor suppressor protein p53. Different levels of Mdm2 are capable of inducing both mono- and polyubiquitination in a dosage dependent manner, thus determining p53's fate. Our findings demonstrate monoubiquitin-mediated protein trafficking can be expanded to nuclear-cytoplasmic shuttling, and also imply similar scenarios may apply to other cellular factors.     

The signaling function of an extra‐floral display: what selects for signal development?

The vertical inflorescences of the Mediterranean annual Salvia viridis carry many small, colorful flowers, and are frequently terminated by a conspicuous tuft of colorful leaves ('flag') that attracts insect visitors. Insects may use the flags as indicators of food rewards in the inflorescences below, as long-distance cues for locating and choosing flowering patches, or both. Clipping of all flags from patches of inflorescences in the field reduced the number of arriving insects, but not the total number of inflorescences and flowers visited by them. The number of flowers visited per inflorescence increased with inflorescence size, and inflorescence and flower visits rates significantly increased with patch size. Six percent of the plants in the study population did not develop any flag during blooming, yet suffered no reduction in seed set as compared to flag-bearing neighboring individuals. Removal of flags from all inflorescences in a patch reduced seed set in comparison with untreated controls, while flag clipping from ten randomly selected inflorescences in a patch did not decrease seed production. These results suggest that flags signal long-distance information to potential pollinators (possibly indicating patch location or size), while flower-related cues may indicate inflorescence quality.
Plants that do not develop flags probably benefit from the flag signals displayed by their neighbors, without bearing the costs of signal production. Greenhouse-grown S. viridis plants allocated a low proportion of their biomass to flags. Plants grown under water stress did not reduce biomass allocation to flags as compared to irrigated controls. Water loss rates of picked flags were lower than those of picked leaves. These findings suggest that the expenses of flag production and maintenance are modest, reducing the selective advantage of individuals that do not carry flags. We discuss additional potential evolutionary mechanisms that may select for flag production.     

β‐Amyloid catabolism: roles for neprilysin (NEP) and other metallopeptidases?

The steady-state level of amyloid beta-peptide (Abeta) represents a balance between its biosynthesis from the amyloid precursor protein (APP) through the action of the beta- and gamma-secretases and its catabolism by a variety of proteolytic enzymes. Recent attention has focused on members of the neprilysin (NEP) family of zinc metalloproteinases in amyloid metabolism. NEP itself degrades both Abeta(1-40) and Abeta(1-42) in vitro and in vivo, and this metabolism is prevented by NEP inhibitors. Other NEP family members, for example endothelin-converting enzyme, may contribute to amyloid catabolism and may also play a role in neuroprotection. Another metalloproteinase, insulysin (insulin-degrading enzyme) has also been advocated as an amyloid-degrading enzyme and may contribute more generally to metabolism of amyloid-forming peptides. Other candidate enzymes proposed include angiotensin-converting enzyme, some matrix metalloproteinases, plasmin and, indirectly, thimet oligopeptidase (endopeptidase-24.15). This review critically evaluates the evidence relating to proteinases implicated in amyloid catabolism. Therapeutic strategies aimed at promoting A,beta degradation may provide a novel approach to the therapy of Alzheimer's disease.     

The choline transporter resurfaces: new roles for synaptic vesicles?

Presynaptic choline uptake is vital to sustained neuronal acetylcholine (ACh) release; however, only with the recent cloning of choline transporters (CHTs) (i.e., SLC5A7), has a picture emerged of the regulatory pathways supporting CHT modulation. Studies arising from the development of CHT-specific antibodies reveal a large, intracellular reserve of CHT proteins, localized to ACh-containing, synaptic vesicles. The intersection of mechanisms supporting vesicular ACh release and choline uptake demonstrates an elegant mechanism for linking regulation of CHT membrane density to rates of ACh release. Furthermore, these studies point to control of the CHT endocytic process as an important target for novel therapeutics that could offset functional deficits in disorders bearing diminished cholinergic tone, including myasthenias and dementias.     

New roles for DIF? Effects on early development in Dictyostelium

The DIFs are unusual, chlorinated molecules which induce stalk cell differentiation during the later, multicellular phase of Dictyostelium development. Here we provide evidence that one or more DIFs have a role during early development, when small amounts are known to be made. Initial indications came from an optical technique which detects changes in shape or cohesion of cells in suspension (Gerisch and Hess, PNAS 71, 2118, 1974). After a period of optical inactivity at the start of development, cell suspensions normally produce spontaneous spike-shaped light-scattering oscillations synchronised by oscillations in extracellular cAMP levels, followed by sinusoidal oscillations where the synchroniser is not known. DIFs 1 and 2 produce optical responses from cells at all these early stages of development. The phase of both spiked and sinusoidal oscillations can be shifted, indicating an effect on the oscillator in each case. We find further: (1) cAMP oscillations and cAMP relay during spiked oscillations are transiently inhibited by DIF-1. (2) DIF-1 causes a transient decrease in cellular cGMP levels in cells taken before oscillations commence and likewise inhibits the cGMP response to a cAMP stimulus in cells taken later in development. Cytoskeletal organization and hence cell shape might be affected by DIF-1 by this indirect route. (3) The effects of DIF-1 are transient, even though it is essentially stable in the cell suspension. Cells somehow adapt to DIF-1. (4) The effects are chemically specific: DIF-1 and DIF-2 are active at 10(-7) to 10(-8) M, with DIF-2 being the more active, whereas related compounds have little or no activity at 10(-6) M. These results indicate that cells are responsive to DIFs 1 and 2 from the start of development and suggest a wider role for the DIFs. This role might involve effects on cAMP signalling and on intracellular second messengers.     

DLK: the "preconditioning" signal for axon regeneration?

In this issue of Neuron, Shin et?al. (2012) show that the dual leucine zipper kinase (DLK) is responsible for the retrograde injury signal in spinal sensory and motor neurons. DLK is required for the accelerated regeneration seen after axotomy and for the improved regeneration seen after a conditioning injury. DLK KO axons have severely reduced axon regeneration in?vivo.