RESUMO
We have previously shown that the RNA-binding protein HuD binds to a regulatory element in the growth-associated protein (GAP)-43 mRNA and that this interaction involves its first two RNA recognition motifs (RRMs). In this study, we investigated the functional significance of this interaction by overexpression of human HuD protein (pcHuD) or its truncated form lacking the third RRM (pcHuD I+II) in PC12 cells. Morphological analysis revealed that pcHuD cells extended short neurites containing GAP-43-positive growth cones in the absence of nerve growth factor (NGF). These processes also contained tubulin and F-actin filaments but were not stained with antibodies against neurofilament M protein. In correlation with this phenotype, pcHuD cells contained higher levels of GAP-43 without changes in levels of other NGF-induced proteins, such as SNAP-25 and tau. In mRNA decay studies, HuD stabilized the GAP-43 mRNA, whereas HuD I+II did not have any effect either on GAP-43 mRNA stability or on the levels of GAP-43 protein. Likewise, pcHuD I+II cells showed no spontaneous neurite outgrowth and deficient outgrowth in response to NGF. Our results indicate that HuD is sufficient to increase GAP-43 gene expression and neurite outgrowth in the absence of NGF and that the third RRM in the protein is critical for this function.
Assuntos
Proteína GAP-43/genética , Regulação da Expressão Gênica , Fator de Crescimento Neural/farmacologia , Proteínas do Tecido Nervoso/metabolismo , Neuritos/fisiologia , Proteínas de Ligação a RNA/metabolismo , Animais , Proteínas ELAV , Proteína Semelhante a ELAV 4 , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas do Tecido Nervoso/genética , Neuritos/efeitos dos fármacos , Neuritos/ultraestrutura , Células PC12 , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/genética , Ratos , Proteínas Recombinantes/metabolismo , Deleção de Sequência , Transcrição Gênica , TransfecçãoRESUMO
The principles of oversampling are exploited in a simple beamforming architecture using a single bit delta-sigma (DeltaC) analog to digital converter (A/D) on every channel. The high sampling rate required for the single bit A/D provides adequate delay accuracy for high quality beamforming using elementary sample manipulations. Images produced with this beamformer exhibit significant artifacts directly related to dynamic focusing. However, a simple digital recording technique following delays permits dynamically focused beamforming without degrading image quality. The simplicity of this beamformer compared to conventional methods may facilitate very large channel count or low power beamformers suitable for 1.5-D arrays or portable scanners.
RESUMO
Delta-sigma (DeltaSigma) modulators can implement a simpler digital ultrasound beamformer than can traditional architectures based on multi-bit analog-to-digital converters (A/D). The signal-to-noise ratio (SNR) of the DeltaSigma modulators, however, suffers from limited oversampling ratios. To improve the SNR of each channel, a mixing signal heterodynes narrowband signals to lower frequencies where the baseband DeltaSigma modulator performs better. Noise figure analyses are presented that illustrate the effectiveness of this technique in improving noise performance. Also, spectral Doppler and color flow simulations are presented that realistically emulate a 32 channel oversampled beamformer and compare these results with traditional and ideal systems.