Arabidopsis thaliana CYCLIC NUCLEOTIDE-GATED CHANNEL2 mediates extracellular ATP signal transduction in root epidermis.

Funder: Agence Nationale de la Recherche; Id: http://dx.doi.org/10.13039/501100001665Funder: Canadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada; Id: http://dx.doi.org/10.13039/501100002790Damage can be signalled by extracellular ATP (eATP) using plasma membrane (PM) receptors to effect cytosolic free calcium ion ([Ca2+ ]cyt ) increase as a second messenger. The downstream PM Ca2+ channels remain enigmatic. Here, the Arabidopsis thaliana Ca2+ channel subunit CYCLIC NUCLEOTIDE-GATED CHANNEL2 (CNGC2) was identified as a critical component linking eATP receptors to downstream [Ca2+ ]cyt signalling in roots. Extracellular ATP-induced changes in single epidermal cell PM voltage and conductance were measured electrophysiologically, changes in root [Ca2+ ]cyt were measured with aequorin, and root transcriptional changes were determined by quantitative real-time PCR. Two cngc2 loss-of-function mutants were used: cngc2-3 and defence not death1 (which expresses cytosolic aequorin). Extracellular ATP-induced transient depolarization of Arabidopsis root elongation zone epidermal PM voltage was Ca2+ dependent, requiring CNGC2 but not CNGC4 (its channel co-subunit in immunity signalling). Activation of PM Ca2+ influx currents also required CNGC2. The eATP-induced [Ca2+ ]cyt increase and transcriptional response in cngc2 roots were significantly impaired. CYCLIC NUCLEOTIDE-GATED CHANNEL2 is required for eATP-induced epidermal Ca2+ influx, causing depolarization leading to [Ca2+ ]cyt increase and damage-related transcriptional response

Spatial origin of the extracellular ATP-induced cytosolic calcium signature in Arabidopsis thaliana roots: wave formation and variation with phosphate nutrition.

Extracellular ATP (eATP) increases cytosolic free calcium ([Ca2+ ]cyt ) as a specific second messenger 'signature' through the plasma membrane DORN1/P2K1 receptor. Previous studies revealed a biphasic signature in Arabidopsis thaliana roots that is altered by inorganic phosphate (Pi) deprivation. The relationship between the two phases of the signature and possible wave formation have been tested as a function of Pi nutrition. The bioluminescent aequorin and intensiometric GCaMP3 reporters were used to resolve the spatial origin of the eATP [Ca2+ ]cyt signature in Arabidopsis root tips. Application of eATP only to the root apex allowed [Ca2+ ]cyt wave resolution without the confounding effects of eATP delivery by superfusion. The first apical millimetre of the root generates the first [Ca2+ ]cyt increase by eATP, regardless of nutritional status. The second increase occurs sub-apically in the root hair zone, has some autonomy and is significantly reduced in Pi-starved roots. A significant component of the Pi-replete signature does not require DORN1/P2K1, but Pi-starved roots appear to have an absolute requirement for that receptor. Application of eATP specifically to the root apex provides evidence for cell-to-cell propagation of a [Ca2+ ]cyt wave that diminishes sub-apically. The apex maintains a robust [Ca2+ ]cyt increase (even under Pi starvation) that is the basis of a propagative wave, with implications for the ability of the root's eATP signalling systems to signal systemically. Partial autonomy of the sub-apical region may be relevant to the perception of eATP from microbes. eATP-induced [Ca2+ ]cyt increase may not have always have an obligate requirement for DORN1/P2K1

Optical proximity correction for 0.13 m SiGe:C BiCMOS

We present results for a rule based optical proximity (RB-OPC) and a model based optical proximity correction (MB-OPC) for 0.13 m SiGe:C BiCMOS technology. The technology provides integrated high performance heterojunction bipolar transistors (HBTs) with cut-off frequencies up to 300 GHz. This requires an optical proximity correction of critical, layers with an excellent mask quality. This paper provides results of the MB-OPC and RB-OPC using the Mentor Calibre software in comparison to uncorrected structures (NO-OPC). We show RB- and MB-OPC methods for the shallow trench and gate layer, and the RB-OPC for the emitter window-, contact- and metal layers. We will discuss the impact of the RB- and MB-OPC rules on the process margin and yield in the 0.13 m SiGe:C BiCMOS technology, based on CD-SEM data obtained from the evaluation of the RB- and MB-OPC corrected SRAM cells