Cross-pathway integration of cAMP signals through cGMP and calcium-regulated phosphodiesterases in mouse striatal cholinergic interneurons.

Background And Purpose: Acetylcholine plays a key role in striatal function. Firing properties of striatal cholinergic interneurons depend on intracellular cAMP through the regulation of I currents. Yet, the dynamics of cyclic nucleotide signalling in these neurons have remained elusive.

Pivotal role of phosphodiesterase 10A in the integration of dopamine signals in mice striatal D and D medium-sized spiny neurones.

Background And Purpose: Dopamine in the striatum plays a crucial role in reward processes and action selection. Dopamine signals are transduced by D and D dopamine receptors which trigger mirror effects through the cAMP/PKA signalling cascade in D and D medium-sized spiny neurons (MSNs). Phosphodiesterases (PDEs), which determine the profile of cAMP signals, are highly expressed in MSNs, but their respective roles in dopamine signal integration remain poorly understood.

Phosphodiesterase 1 Bridges Glutamate Inputs with NO- and Dopamine-Induced Cyclic Nucleotide Signals in the Striatum.

The calcium-regulated phosphodiesterase 1 (PDE1) family is highly expressed in the brain, but its functional role in neurones is poorly understood. Using the selective PDE1 inhibitor Lu AF64196 and biosensors for cyclic nucleotides including a novel biosensor for cGMP, we analyzed the effect of PDE1 on cAMP and cGMP in individual neurones in brain slices from male newborn mice. Release of caged NMDA triggered a transient increase of intracellular calcium, which was associated with a decrease in cAMP and cGMP in medium spiny neurones in the striatum.

Quantitative analysis of the interaction between l-methionine derivative and oligonucleotides.

This study explores the use of l-methionine derivative as a potential affinity ligand for nucleic acids purification. The l-methionine derivative is synthesized by activation of the carboxylic acid group with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide follow by immobilization on amine sensor surface, previously activated and treated with ethylenediamine. Their affinity towards oligonucleotides has been determined by surface plasmon resonance biosensor.

Quantitative analysis of histamine- and agmatine-DNA interactions using surface plasmon resonance.

To exploit the binding affinity for efficient plasmid purification, agmatine and histamine were immobilized on carboxymethylated dextran surface. Their binding strength is evaluated with oligonucleotides and pUC19 (2.69 kbp), pVAX1-LacZ (6.

Rapid quantification of supercoiled plasmid deoxyribonucleic acid using a monolithic ion exchanger.

The demand for high-purity supercoiled plasmid DNA to be applied as a vector for new therapeutic strategies, such as gene therapy or DNA vaccination has increased in the last years. Thus, it is necessary to implement an analytical technique suitable to control the quality of the supercoiled plasmid as a pharmaceutical product during the manufacturing process. The present study describes a new methodology to quantify and monitor the purity of supercoiled plasmid DNA by using a monolithic column based on anion-exchange chromatography.