Altered expression of GABAA receptors (α4, γ2 subunit), potassium chloride cotransporter 2 and astrogliosis in tremor rat hippocampus

Abstract Objective Today, the research about the involvement of non-coding RNAs on neurological disorders is still scarce. Hence, we aimed to investigate the effect of miR-96 targeted RAC1 to mediate RhoA/ROCK signaling pathway in neurological damage in acute seizure rats.MethodsThe acute seizure rat model was constructed by using classical chlorinated-pilocarpine injection, which was treated with miR-96 mimics, RAC1-siRNA or their controls. The number of BrdU positive cells, neuronal changes and apoptosis, expression of NGF, BDNF and GFAP were detected by different staining assays. At the same time, the activity of SOD, the content of MDA and the protein contents of TNF-α and IL-6 in rat hippocampus were detected as well. Next, the expression of NGF, BDNF, GFAP, Bax, Bcl-2, caspase-3, RAC1, RhoA and ROCK were determined by RT-qPCR and western blot analysis. Then, the binding site between miR-96 and RAC1 was analyzed by bioinformatics software and luciferase assay. Finally, the altered expression of miRNAs was investigated in exosomes released from excitotoxic neurons with a customized rat miRNA chipset.Results After upregulation of miR-96 or downregulation RAC1, BrdU-positive cells, TUNEL-positive cells, NGF, GFAP, MDA, TNF-α, IL-6, Bax, caspase-3, and pathway related proteins in rat hippocampus decreased while Bcl-2, BDNF, and the activity of SOD increased. Furthermore, RAC1 was found to be the target gene of miR-96 and their relationship was validated in in-vitro studies. Finally, the decreased expression of miR-96 in exosomes from kainic acid treated neurons was identified as well.Conclusion This study suggests that upregulation of miR-96 could downregulate RAC1 and inhibit the activation of RhoA/ROCK signaling pathway, thus alleviated the neurological damage in acute seizure rats.

Download Full-text

Loop diuretics: from the Na-K-2Cl transporter to clinical use

AJP Renal Physiology ◽

10.1152/ajprenal.00119.2002 ◽

2003 ◽

Vol 284 (1) ◽

pp. F11-F21 ◽

Cited By ~ 108

Author(s):

Sudha S. Shankar ◽

D. Craig Brater

Keyword(s):

Potassium Chloride ◽

Loop Diuretics ◽

Loop Of Henle ◽

Gene Encoding ◽

Sodium Potassium ◽

Altered Expression ◽

Disease States ◽

Expression Activity ◽

Diuretic Response ◽

Renal Transporter

The diuretic response to loop diuretics in various disease states has consistently been found to be subnormal. One of the key determinants of the degree of diuretic response is the functional integrity of the sodium-potassium-chloride transporter in the loop of Henle. Studies in animal models suggest that expression/activity of the transporter may be affected by factors such as altered natural splicing events of NKCC2 (the gene encoding for the renal transporter), renal prostanoids, vasopressin, and other autacoids. We have reviewed the pharmacokinetics and pharmacodynamics of loop diuretics in health and in edematous disorders for which they are used. On the basis of evidence reviewed in this paper, we propose that altered expression or activity of the sodium-potassium-chloride transporter in the loop of Henle, in conjunction with events occurring in other segments of the nephron, possibly accounts for the altered diuretic response to these agents. Thus the modulators of this altered expression/activity could serve as important therapeutic targets for alternative diuretic regimens in these conditions.

Download Full-text