Transverse Vein Differentiation Associated with Gas Space Formation--Fate of the Middle Cell Layer in Leaf Sheath Development of Rice

In monocotyledons, the leaf vascular network consists of a hierarchicalsequence of vertical vascular bundles and numerous transverseveins that interconnect adjacent vertical veins. In the leafsheath of these species, especially grasses, lysigenous gascavities (gas spaces) are developed into intervascular spacesand provide a gas conducting system to non-aerial parts underflooded conditions. The spatial relationship between gas spaceformation and transverse vein differentiation was investigatedusing the leaf sheath of rice (Oryza sativa L.). Histochemicalobservation showed that patterns of differentiation of the transversevein are distinct from those of vertical vascular bundles. Onthe other hand, gas spaces are formed through the processesof cell death (collapse). Both events are initiated at a specificcell position in the middle layers of the leaf sheath, fromwhich the vascular system of the leaf is derived; this indicatesthat differentiation of transverse veins is associated withgas space formation. The cell-to-cell movement of fluoresceinisothiocyanate-conjugated dextran injected into middle layercells coincided with the area where cell collapse occurred,indicating a close relationship between the middle and adaxialcell layers, but not abaxial cell layers. A uniform cell numberbetween each transverse vein in the leaf sheath suggested theinvolvement of spatial regulation in transverse vein formationregardless of clonal history at the later stage of leaf veincanalization. Copyright 2000 Annals of Botany Company Cell collapse, leaf development, middle cell layer, microinjection, Oryza sativa L., rice, programmed cell death.     

Seed Coat Microsculpturing in Brassica and Allied Genera (Subtribes Brassicinae, Raphaninae, Moricandiinae)

To obtain new information on the taxonomy of Brassica and alliedgenera, seed surfaces of 44 species (78 accessions) belongingto 11 genera of subtribes Brassicinae, Raphaninae and Moricandiinaewere examined using a scanning electron microscope. Ten typesof basic ornamentation patterns were recognized. While fourtypes were represented by only one species each, six types hadmore than one species either representing one genus (two types)or more (four types). The different species in each of the sixtypes could generally be distinguished from each other on thebasis of differences in microsculpturing features. This analysisprovides evidence for the close relationship among the variousgenera within the subtribe Brassicinae, and also the closenessof Raphanus, Enarthrocarpus and Moricandia of subtribe Raphaninaeand Moricandiinae, respectively, with the Brassicinae. Copyright2000 Annals of Botany Company Brassicinae, Moricandiinae, Raphaninae, SEM, seed coat microsculpturing, taxonomic implications     

Role of the salt-bridge between switch-1 and switch-2 of Dictyostelium myosin.

Motifs N2 and N3, also referred to as switch-1 and switch-2, form part of the active site of molecular motors such as myosins and kinesins. In the case of myosin, N3 is thought to act as a gamma-phosphate sensor and moves almost 6 A relative to N2 during the catalysed turnover of ATP, opening and closing the active site surrounding the gamma-phosphate. The closed form seems to be necessary for hydrolysis and is stabilised by the formation of a salt-bridge between an arginine residue in N2 and a glutamate residue in N3. We examined the role of this salt-bridge in Dictyostelium discoideum myosin. Myosin motor domains with mutations E459R or R238E, that block salt-bridge formation, show defects in nucleotide-binding, reduced rates of ATP hydrolysis and a tenfold reduction in actin affinity. Inversion of the salt-bridge in double-mutant M765-IS eliminates most of the defects observed for the single mutants. With the exception of a 2,500-fold higher KMvalue for ATP, the double-mutant displayed enzymatic and functional properties very similar to those of the wild-type protein. Our results reveal that, independent of its orientation, the salt-bridge is required to support efficient ATP hydrolysis, normal communication between different functional regions of the myosin head, and motor function.     

Integrins enhance platelet-derived growth factor (PDGF)-dependent responses by altering the signal relay enzymes that are recruited to the PDGF beta receptor.

Since the extracellular matrix (ECM) can promote platelet-derived growth factor (PDGF)-dependent responses, we hypothesized that the ECM mediates this effect by preventing the PDGF beta receptor (betaPDGFR) from associating with the negative regulator, RasGAP (the GTPase-activating protein of Ras). We found that binding of RasGAP to the wild-type betaPDGFR was decreased; the activation of Ras and Erk was enhanced, and [3H]thymidine uptake was better in cells cultured on fibronectin than in cells cultured on polylysine. To investigate the mechanism by which culturing cells on fibronectin diminished the recruitment of RasGAP to the betaPDGFR, we focused on SHP-2 since it dephosphorylates the betaPDGFR at the phosphotyrosine required for binding of RasGAP. Culturing cells on fibronectin increased the amount of SHP-2 that associated with the betaPDGFR. Furthermore, cells expressing receptor mutants that failed to associate with SHP-2 were insensitive to fibronectin. The ECM enhances PDGF-dependent responses by increasing the association of SHP-2 with the betaPDGFR, which in turn decreases the time that RasGAP interacts with the receptor. Thus, fibronectin changes PDGF-dependent signaling and biological responses by altering the signal relay enzymes that are recruited to the receptor.     

Field Hypothesis on the Self-regulation of Gene Expression

The mechanism of the self-regulation of gene expression in living cells is generally explained by considering complicated networks of key-lock relationships, and in fact there is a large body of evidence on a hugenumber of key-lock relationships. However, in the present article we stress that with the network hypothesis alone it is impossible to fully explain the mechanism of self-regulation in life. Recently, it has been established that individual giant DNA molecules, larger than several tens of kilo base pairs, undergo a large discrete transition in their higher-order structure. It has become clear that nonspecific weak interactions with various chemicals, suchas polyamines, small salts, ATP and RNA, cause on/off switching in the higher-order structure of DNA. Thus, the field parameters of the cellular environment should play important roles in the mechanism of self-regulation, in addition to networks of key and locks. This conformational transition induced by field parameters may be related to rigid on/off regulation, whereas key-lock relationships may be involved in a more flexible control of gene expression.     

Obesity-inducing lesions of the central nervous system alter leptin uptake by the blood-brain barrier.

Leptin regulates body adiposity by decreasing feeding and increasing thermogenesis. Obese humans and some obese rodents are resistant to peripherally administered leptin, suggesting a defect in the transport of leptin across the blood-brain barrier (BBB). Defective transport of exogenous leptin occurs in some models of obesity, but in other models transport is normal. This shows that factors other than obesity are associated with impairment of leptin transport across the BBB. In order to further investigate these factors, we determined leptin transport in rats made obese by lesioning of the ventromedial hypothalamus (VMH), paraventricular nucleus (PVN), or posterodorsal amygdala (PDA). These regions all contain leptin receptors and lesions there induce obesity and hyperleptinemia and alter the levels of many feeding hormones which might participate in leptin transporter regulation. We measured the uptake of radioactively labeled leptin by the BBB by multiple-time regression analysis which divides uptake into a reversible phase (Vi, e.g., receptor/transporter binding to the brain endothelial cell) and an irreversible phase (Ki, complete transport across the BBB). Leptin uptake was not affected in rats with VMH lesions. No significant change occurred in the entry rate (Ki) for any group, although Ki declined by over 35% in rats with PVN lesions. Decreased uptake was observed in rats with PVN lesions and with PDA lesions. This was primarily due to a reduced Vi (about 21% for the PDA). This decreased uptake is most likely explained by decreased binding of leptin to the brain endothelial cell, which could be because of decreased binding by either receptors or transporters. This suggests that some of the feeding hormones controlled by the PVN and PDA may participate in regulating leptin uptake by the BBB.