Ca 2؉ signals are thought to play important roles in plant growth and development, including key aspects of pollen tube growth and fertilization. The dynamics of a Ca 2؉ signal are largely controlled by influx (through channels) and efflux (through pumps and antiporters). The Arabidopsis genome encodes 14 Ca 2؉ pumps, 10 of which belong to a family of autoinhibited Ca 2؉ ATPases (ACA) that are predicted to be activated by Ca 2؉ ͞calmodulin. Here, we show that isoform ACA9 is expressed primarily in pollen and localized to the plasma membrane. Three independent T-DNA [portion of the Ti (tumor-inducing) plasmid that is transferred to plant cells] gene disruptions of ACA9 were found to result in partial male sterility. Complementation was observed by using a ACA9-yellow fluorescence protein (YFP) fusion that displayed plasma membrane localization. Mutant aca9 pollen displayed a reduced growth potential and a high frequency of aborted fertilization, resulting in a >80% reduction in seed set. These findings identify a plasma membrane Ca 2؉ transporter as a key regulator of pollen development and fertilization in flowering plants.

Calcium-dependent protein kinases (CDPKs) are specific to plants and some protists. Their activation by calcium makes them important switches for the transduction of intracellular calcium signals. Here, we identify the subcellular targeting potentials for nine CDPK isoforms from Arabidopsis, as determined by expression of green fluorescent protein (GFP) fusions in transgenic plants. Subcellular locations were determined by fluorescence microscopy in cells near the root tip. Isoforms AtCPK3-GFP and AtCPK4-GFP showed a nuclear and cytosolic distribution similar to that of free GFP. Membrane fractionation experiments confirmed that these isoforms were primarily soluble. A membrane association was observed for AtCPKs 1, 7, 8, 9, 16, 21, and 28, based on imaging and membrane fractionation experiments. This correlates with the presence of potential N-terminal acylation sites, consistent with acylation as an important factor in membrane association. All but one of the membrane-associated isoforms targeted exclusively to the plasma membrane. The exception was AtCPK1-GFP, which targeted to peroxisomes, as determined by covisualization with a peroxisome marker. Peroxisome targeting of AtCPK1-GFP was disrupted by a deletion of two potential N-terminal acylation sites. The observation of a peroxisome-located CDPK suggests a mechanism for calcium regulation of peroxisomal functions involved in oxidative stress and lipid metabolism.

Ion signals are critical to regulating polarized growth in many cell types, including pollen in plants and neurons in animals. Genetic evidence presented here indicates that pollen tube growth requires cyclic nucleotide-gated channel (CNGC) 18. CNGCs are nonspecific cation channels found in plants and animals and have well established functions in excitatory signal transduction events in animals. In Arabidopsis, male sterility was observed for two cngc18 null mutations. CNGC18 is expressed primarily in pollen, as indicated from a promoter::GUS (␤-glucuronidase) reporter analysis and expression profiling. The underlying cause of sterility was identified as a defect in pollen tube growth, resulting in tubes that were kinky, short, often thin, and unable to grow into the transmitting tract. Expression of a GFP-tagged CNGC18 in mutant pollen provided complementation and evidence for asymmetric localization of CNGC18 to the plasma membrane at the growing tip, starting at the time of pollen grain germination. Heterologous expression of CNGC18 in Escherichia coli resulted in a time-and concentration-dependent accumulation of more Ca 2؉ . Thus, CNGC18 provides a mechanism to directly transduce a cyclic nucleotide (cNMP) signal into an ion flux that can produce a localized signal capable of regulating the pollen tip-growth machinery. These results identify a CNGC that is essential to an organism's life cycle and raise the possibility that CNGCs have a widespread role in regulating cell-growth dynamics in both plant and animals.calcium ͉ male sterile ͉ Arabidopsis ͉ CaM

This study demonstrates a significant risk of serious neurologic injury after cervical TF-ESIs. A growing body of evidence supports an embolic mechanism, whereby inadvertent intra-arterial injection of particulate corticosteroid causes a distal infarct. Embolism to the distal basilar artery region can cause midbrain, pons, cerebellum, thalamus, temporal and occipital lobe infarctions. Other potential mechanisms of infarction include vertebral artery perforation causing dissection/thrombosis and needle-induced vasospasm.

SUMMARYCalcium signals are critical for the regulation of polarized growth in many eukaryotic cells, including pollen tubes and neurons. In plants, the regulatory pathways that code and decode Ca 2+ signals are poorly understood. In Arabidopsis thaliana, genetic evidence presented here indicates that pollen tube tip growth involves the redundant activity of two Ca 2+ -dependent protein kinases (CPKs), isoforms CPK17 and -34. Both isoforms appear to target to the plasma membrane, as shown by imaging of CPK17-yellow fluorescent protein (YFP) and CPK34-YFP in growing pollen tubes. Segregation analyses from two independent sets of T-DNA insertion mutants indicate that a double disruption of CPK17 and -34 results in an approximately 350-fold reduction in pollen transmission efficiency. The near sterile phenotype of homozygous double mutants could be rescued through pollen expression of a CPK34-YFP fusion. In contrast, a transgene rescue was blocked by mutations engineered to disrupt the Ca 2+ -activation mechanism of CPK34 (CPK34-YFP-E465A,E500A), providing in vivo evidence linking Ca 2+ activation to a biological function of a CPK. While double mutant pollen tubes displayed normal morphology, relative growth rates for the most rapidly growing tubes were reduced by more than three-fold compared with wild type. In addition, while most mutant tubes appeared to grow far enough to reach ovules, the vast majority (>90%) still failed to locate and fertilize ovules. Together, these results provide genetic evidence that CPKs are essential to pollen fitness, and support a mechanistic model in which CPK17 and -34 transduce Ca 2+ signals to increase the rate of pollen tube tip growth and facilitate a response to tropism cues.