Friday, May 13, 2011

PKA+siRNA Block Hyperalgesia

I have reported use of our i-FectTM siRNA delivery kit for gene expression analysis studies of DOR, hTERT, The β3 subunit of the Na+,K+-ATPase, rSNSR1, NTS1. NAV1.8, , TRPV1, Survivin, Flaviviruses and more.

The data in this pub indicates that selective knock-down of spinal PKA activity by intrathecal ( pretreatment of rats with a PKA-selective small interference RNA (siRNA) mixture significantly attenuates sustained morphine-mediated augmentation of spinal CGRP immunoreactivity, thermal hyperalgesia, mechanical allodynia and antinociceptive tolerance. The present findings indicate that sustained morphine-mediated activation of spinal cAMP/PKA-dependent signaling may play an important role in opioid induced hyperalgesia: S. Tumati, W.R. Roeskea, T.M. Largent-Milnesa, T.W. Vanderaha, and EV Varga. Intrathecal PKA-selective siRNA treatment blocks sustained morphine-mediated pain sensitization and antinociceptive tolerance in rats. doi:10.1016/j.jneumeth.2011.04.036.

Figure: Intrathecal PKA-selective siRNA treatment blocks the development of morphine antinociceptive tolerance.
Male Sprague Dawley rats were pretreated with vehicle (inverted triangle) or PKA-selective siRNA (circle) for 3 days. After the pretreatments, the animals received continuous saline (open symbols, error bars within the symbol) or morphine (45 nmol/μl/h) (closed symbols, error bars within the symbol) infusion for 6 days, with continued siRNA or vehicle injections on alternate days. Sustained (6 days) systemic morphine (45 nmol/μl/h) infusion caused a rightward shift in the dose-response curve, with the previous A90 dose causing only 20±1% MPE (**p < 0.01 relative to control, one-way ANOVA, n=5). Intrathecal PKAselective siRNA pre-treatment greatly attenuated sustained morphine-mediated rightward shift in the morphine dose-response curve. Thus, re-challenge with the naive A90 dose (10 μg/5μl) produced 93±2% antinociception in the PKA-selective siRNA pre-treated rat**p < 0.01 relative to vehicle pre-treated morphine-infused rats, one-way ANOVA, n=5). 

Transfection Kits and Related Reagents:

i-Fect ™
-A novel cationic  lipid formulation specifically designed for efficient delivery of 27mer DsiRNAs(dicer substrate small Interfering RNAs)& 21mer siRNAs (small interfering RNAs) in vitro and in vivo.
-A cationic lipid that has been specifically formulated for nervous system  applications. n-Fect provides higher transfection efficiency than  other commercially available broad-spectrum transfection reagents for glial cells, neuronal cell lines, and certain primary neuronal  cultures.
-A broad-spectrum  transfection reagent successfully used in many cell types commonly used by neuroscientist.
 pn-Fect™ -The latest advance in transfection technology for primary neuronal  cells. This unique reagent provides ultra-high plasmid DNA delivery efficiencies and low cytotoxicity compared to competitive reagents.
-Designed to delivery plasmids, DNA or RNA to hard to transfect Cell Lines.
-Is a unique  lipid-based formulation that allows the delivery of proteins,  peptides or other bioactive molecules into a broad range of cell  types.
-A peptide for  delivering small molecules into Neurons and other cells. MP  Biomedical is the manufacturer of Penetratin-1
MATra™ Products
-Provides a system for Magnetically Driving the transfection process enhancing
the performance of transfectants.

Other Cells
-Competent mammalian cells by Category
Primary Neurons and Astrocytes

I'll be posting more soon.

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