Neurite outgrowth resistance to rho kinase inhibitors in PC12 Adh cell

Nicotinic acetylcholine receptors (nAChRs) modulate the growth and structure of neurons throughout the nervous system. Ligand stimulation of the a7 nAChR has been shown to regulate the large heterotrimeric GTP-binding protein (G protein) signaling in various types of cells. Here, we demonstrate a role for a7 nAChR/G protein interaction in the activation of the small (monomeric) RhoA GTPase leading to cytoskeletal changes during neurite growth. Treatment of PC12 cells with the a7 nAChR agonist choline or PNU-282987 was associated with an increase in RhoA activity and an inhibition in neurite growth. Specifically, choline treatment was found to attenuate the velocity of microtubule growth at the growth cone and decrease the rate of actin polymerization throughout the cell. The effects of a7 nAChR activation were abolished by expression of a dominant negative a7 nAChR (a7 345-348A ) deficient in G protein coupling. Proteomic analysis of immunoprecipitated a7 nAChR complexes from differentiating PC12 cells and synaptic fractions of the developing mouse hippocampus revealed the existence of Rho GTPase-regulating guanine nucleotide exchange factors within a7 nAChR interactomes. These findings underscore the role of a7 nAChR/G protein in cytoskeletal regulation during neurite growth.

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“…; Yin et al . ). We assessed the effects of choline on Rho GTPase expression during PC12 differentiation.…”

Section: Resultsmentioning

confidence: 97%

“…; Yin et al . ). RhoA has also been shown to regulate the polymerization of actin (Nobes and Hall ), play a role in the regulation of the microtubule cytoskeleton (Nobes and Hall ; Etienne‐Manneville and Hall ; Duman et al .…”

Section: Discussionmentioning

confidence: 97%

Alpha 7 nicotinic receptor coupling to heterotrimeric G proteins modulates RhoA activation, cytoskeletal motility, and structural growth

King

Kabbani

2016

Journal of Neurochemistry

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“…RNA quantity and quality were measured by the NanoDrop ND-1000 spectrophotometer. RNA integrity was assessed by standard denaturing agarose gel electrophoresis [ 7 , 18 ].…”

Section: Methodsmentioning

confidence: 99%

Intestinal tract is an important organ for lowering serum uric acid in rats

Yin

Gao

et al. 2017

PLoS ONE

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The kidney was recognized as a dominant organ for uric acid excretion. The main aim of the study demonstrated intestinal tract was an even more important organ for serum uric acid (SUA) lowering. Sprague-Dawley rats were treated normally or with antibiotics, uric acid, adenine, or inosine of the same molar dose orally or intraperitoneally for 5 days. Rat’s intestinal tract was equally divided into 20 segments except the cecum. Uric acid in serum and intestinal segment juice was assayed. Total RNA in the initial intestinal tract and at the end ileum was extracted and sequenced. Protein expression of xanthine dehydrogenase (XDH) and urate oxidase (UOX) was tested by Western blot analysis. The effect of oral UOX in lowering SUA was investigated in model rats treated with adenine and an inhibitor of uric oxidase for 5 days. SUA in the normal rats was 20.93±6.98 μg/ml, and total uric acid in the intestinal juice was 308.27±16.37 μg, which is two times more than the total SUA. The uric acid was very low in stomach juice, and attained maximum in the juice of the first segment (duodenum) and then declined all the way till the intestinal end. The level of uric acid in the initial intestinal tissue was very high, where XDH and most of the proteins associated with bicarbonate secretion were up-regulated. In addition, SUA was decreased by oral UOX in model rats. The results suggested that intestinal juice was an important pool for uric acid, and intestinal tract was an important organ for SUA lowering. The uric acid distribution was associated with uric acid synthesis and secretion in the upper intestinal tract, and reclamation in the lower.

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“…The RNA quantity and quality were measured by a NanoDrop ND-1000 spectrophotometer. RNA integrity was assessed by standard denaturing agarose gel electrophoresis (denatured by 0.8% methanal), and three bands in one lane suggested its integrity (Chen et al, 2017;Yin et al, 2015).…”

Section: Rt-pcr and High Throughput Sequencingmentioning

confidence: 99%

Uricase-deficient rat is generated with CRISPR/Cas9 technique

Zhang

Dong

et al. 2020

PeerJ

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Urate oxidase (uricase, Uox) is a big obstacle for scientists to establish stable animal models for studying hyperuricemia and associated disorders. Due to the low survival rate of uricase-deficient mice, we generated a Uox-knockout model animal from Sprague Dawley (SD) rats using the CRISPR/Cas9 technique by deleting exons 2 to 4 of the Uox gene. The uricase-deficient rats were named “Kunming-DY rats”, and were apparently healthy with more than a 95% survival up to one year. The male rats’ serum uric acid (SUA) increased to 48.3 ± 19.1 µg/ml, significantly higher than those of wild-type rats. Some indexes of the blood fat like total triglyceride, low density lipoprotein, and renal function indexes including blood urea nitrogen and serum creatinine were significantly different from those of wild-type rats, however, all the indexes were close to or in normal ranges. Histological renal changes including mild glomerular/tubular lesions were observed in these uricase-deficient rats. Thus, “Kunming-DY rats” with stable uricase-deficiency were successfully established and are an alternative model animal to study hyperuricemia and associated diseases mimicking human conditions.

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Neurite outgrowth resistance to rho kinase inhibitors in PC12 Adh cell

Cited by 16 publications

References 36 publications

Alpha 7 nicotinic receptor coupling to heterotrimeric G proteins modulates RhoA activation, cytoskeletal motility, and structural growth

Alpha 7 nicotinic receptor coupling to heterotrimeric G proteins modulates RhoA activation, cytoskeletal motility, and structural growth

Intestinal tract is an important organ for lowering serum uric acid in rats

Uricase-deficient rat is generated with CRISPR/Cas9 technique

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