Distribution of Rho-Kinase in the Bovine Brain

doi:10.1006/bbrc.1999.1409

Biochemical and Biophysical Research Communications

Volume 263, Issue 2, 24 September 1999, Pages 575-579

https://doi.org/10.1006/bbrc.1999.1409 Get rights and content

Abstract

Rho-associated kinase (Rho-kinase) is a serine/threonine protein kinase downstream of the small GTPase Rho, which participates in signaling pathways of many cellular functions. Although Rho-kinase is implicated in the regulation of the morphology of neuronal cells, the distribution of Rho-kinase in the brain has not been elucidated yet. In this study, we investigated the distribution of Rho-kinase using three antibodies recognizing the different epitopes of Rho-kinase. Rho-kinase was abundantly expressed in the gray matter in comparison with the white matter. Strong immunoreactivity was observed in the pyramidal neurons of the cerebral cortex and hippocampus and in the Purkinje cells of the cerebellum. These results indicate that Rho-kinase is abundantly distributed in neurons and might play an important role in remodeling of neurites.

References (30)

Y. Takai et al.
Trends Biochem. Sci.
(1995)
T. Ishizaki et al.
FEBS Lett.
(1997)
T. Leung et al.
J. Biol. Chem.
(1995)
O. Nakagawa et al.
FEBS Lett.
(1996)
M. Amano et al.
J. Biol. Chem.
(1996)
H. Kosako et al.
J. Biol. Chem.
(1997)
R. Hashimoto et al.
Biochem. Biophys. Res. Commun.
(1998)
H. Goto et al.
J. Biol. Chem.
(1998)
H. Inada et al.
Biochem. Biophys. Res. Commun.
(1998)
H. Katoh et al.
J. Biol. Chem.
(1998)

C. Olenik et al.

Brain Res. Mol. Brain Res.

(1997)

T. Hashimoto et al.

Brain Res. Mol. Brain Res.

(1998)

B. Seidel et al.

Brain Res.

(1995)

A. Hall

Science

(1998)

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Alcohol misuse persists as a prevalent societal concern and precipitates diverse deleterious consequences, entailing significant associated health hazards including acute alcohol intoxication (AAI). Binge drinking, a commonplace pattern of alcohol consumption, may incite neurodegeneration and neuronal dysfunction. Clinicians tasked with managing AAI confront a dearth of pharmaceutical intervention alternatives. In contrast, natural products have garnered interest due to their compatibility with the human body and fewer side effects. Lingjiao Gouteng decoction (LGD), a classical traditional Chinese medicine decoction, represents a frequently employed prescription in cases of encephalopathy, although its efficacy in addressing acute alcoholism and alcohol-induced brain injury remains inadequately investigated.
To investigate the conceivable therapeutic benefits of LGD in AAI and alcohol-induced brain injury, while delving into the underlying fundamental mechanisms involved.
We established an AAI mouse model through alcohol gavage, and LGD was administered to the mice twice at the 2 h preceding and 30 min subsequent to alcohol exposure. The study encompassed the utilization of the loss of righting reflex assay, histopathological analysis, enzyme-linked immunosorbent assays, and cerebral tissue biochemical assays to investigate the impact of LGD on AAI and alcohol-induced brain injury. These assessments included a comprehensive evaluation of various biomarkers associated with the inflammatory response and oxidative stress. Finally, RT-qPCR, Western blot, and immunofluorescence staining were carried out to explore the underlying mechanisms through which LGD exerts its therapeutic influence, potentially through the regulation of the RhoA/ROCK2/NF-κB signaling pathway.
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The present study confirmed the therapeutic potential of LGD in AAI and alcohol-induced brain injury, and the protective effects of LGD against alcohol-induced brain injury may be intricately linked to the RhoA/ROCK2/NF-κB signaling pathway.
The role of RhoA/ROCK pathway in the ischemic stroke-induced neuroinflammation
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It is widely known that ischemic stroke is the prominent cause of death and disability. To date, neuroinflammation following ischemic stroke represents a complex event, which is an essential process and affects the prognosis of both experimental stroke animals and stroke patients. Intense neuroinflammation occurring during the acute phase of stroke contributes to neuronal injury, BBB breakdown, and worse neurological outcomes. Inhibition of neuroinflammation may be a promising target in the development of new therapeutic strategies. RhoA is a small GTPase protein that activates a downstream effector, ROCK. The up-regulation of RhoA/ROCK pathway possesses important roles in promoting the neuroinflammation and mediating brain injury. In addition, nuclear factor-kappa B (NF-κB) is another vital regulator of ischemic stroke-induced neuroinflammation through regulating the functions of microglial cells and astrocytes. After stroke onset, the microglial cells and astrocytes are activated and undergo the morphological and functional changes, thereby deeply participate in a complicated neuroinflammation cascade. In this review, we focused on the relationship among RhoA/ROCK pathway, NF-κB and glial cells in the neuroinflammation following ischemic stroke to reveal new strategies for preventing the intense neuroinflammation.
Investigations on Rho/ROCK signaling in post-traumatic stress disorder-like behavior in mice
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Post-Traumatic Stress Disorder (PTSD) is a chronic condition that occurs in response to a traumatic event, and consequently, enhances the threat sensitivity. Rho/ROCK signaling has been implicated in the consolidation of fear memory, stress, depression, anxiety, and traumatic brain injury. However, its role in post-traumatic stress disorder remains elusive. Therefore, the present study was designed to explore the role of fasudil, a Rho/ROCK inhibitor, a mouse model of PTSD. Mice were subjected to underwater trauma stress followed by three situational reminders. Underwater trauma (UWT) significantly increased the freezing behavior, a marker of the formation of aversive memory, in response to situational reminders on the 3rd, 7th, and 14th days, suggesting the significant development of PTSD. Trauma and situational reminders were also associated with significant changes in behavioral parameters in open field, social interaction and actophotometer tests, along with a reduction in serum corticosterone levels. Fasudil (10 and 15 mg/kg) and sertraline (15 mg/kg), a standard drug for PTSD, significantly decreased the freezing behaviour in response to situational reminders, suggesting the inhibition of the formation of aversive fear memory. However, fasudil and sertraline did not modulate normal memory functions, as assessed on elevated plus maze test, before subjecting mice to traumatic stress. Treatment with fasudil and sertraline significantly restored the behavioral changes and normalized the corticosterone levels. Fasudil-mediated blockade of the Rho/ROCK pathway may be responsible for blocking the formation of aversive memory during the traumatic event, which may be manifested in form of decreased contextual fear response during situational reminders.
Increased expression of Rho-associated protein kinase 2 confers astroglial Stat3 pathway activation during epileptogenesis
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Citation Excerpt :
The reactive astrocytosis may facilitate the development of epileptic seizures (de Lanerolle et al., 2010). The cells of the high ROCK2 expression were found in pyramidal neurons of the hippocampus and cerebral cortex and Purkinje cells of the cerebellum (Hashimoto et al., 1999). By immuno-fluorescence, ROCK2 was mainly localized in the neurons in our study.
Patients with TLE are prone to tolerance to antiepileptic drugs. Based on the perspective of molecular targets for drug resistance, it is necessary to explore effective drug resistant genes and signaling pathways for the treatment of TLE. We performed gene expression profiles in hippocampus of patients with drug-resistant TLE and identified ROCK2 as one of the 20 most significantly increased genes in hippocampus. In vitro and in vivo experiments were performed to identify the potential role of ROCK2 in epileptogenesis. In addition, the activity of Stat3 pathway was tested in rat hippocampal tissues and primary cultured astrocytes. The expression levels of ROCK2 in the hippocampus of TLE patients were significantly increased compared with the control group, which was due to the hypomethylation of ROCK2 promoter. Fasudil, a specific Rho-kinase inhibitor, alleviated epileptic seizures in the pilocarpine rat model of TLE. Furthermore, ROCK2 activated the Stat3 pathway in pilocarpine-treated epilepsy rats, and the spearman correlation method confirmed that ROCK2 is associated with Stat3 activation in TLE patients. In addition, ROCK2 was predominantly expressed in astrocytes during epileptogenesis, and induced epileptogenesis by activating astrocyte cell cycle progression via Stat3 pathway. The overexpressed ROCK2 plays an important role in the pathogenesis of drug-resistant epilepsy. ROCK2 accelerates astrocytes cell cycle progression via the activation of Stat3 pathway likely provides the key to explaining the process of epileptogenesis.
Antidepressant-like effects of water extract of Cordyceps militaris (Linn.) Link by modulation of ROCK2/PTEN/Akt signaling in an unpredictable chronic mild stress-induced animal model
2021, Journal of Ethnopharmacology
Citation Excerpt :
Rho-associated protein kinase (ROCK) plays an important role in neuronal survival (Lingor et al., 2008), and it has two isoforms: ROCK1 and ROCK2. The latter is extensively expressed in the central nervous system, including the brain (Hashimoto et al., 1999). The ROCK2 activity involves the phosphatase and tensin homolog (PTEN) and protein kinase B (Akt), which are closely associated with neuronal survival and degeneration.
Cordyceps militaris (Linn.) Link (CM) is a medicinal mushroom traditionally used in tonics for treating several neurological disorders, including epilepsy and anxiety, in Asia. Reports have shown that CM has anti-inflammatory and anti-oxidative effects and may be beneficial for depression management.
This study aimed to investigate the potential of CM as an antidepressant for a long-term unpredictable chronic mild stress (UCMS) rodent models and explore its underlying mechanisms.
Rats were orally administered with 125 (low, L), 250 (medium, M), and 500 (high, H) mg/kg bodyweight (bw) of the water extract of CM (WCM) for 35 consecutive days in the UCMS protocol. The levels of cerebral serotonin (5-HT), dopamine (DA), and metabolites in the frontal cortex of the rats were measured. Blood was collected to investigate the levels of proinflammatory cytokines, and the brain was dissected to assay the stress-associated ROCK2/PTEN/Akt signaling.
All doses of the WCM prevented abnormal behaviors induced by UCMS, including anhedonia and hypoactivity. The LWCM treatment reduced the turnover rate of 5-HT, and all doses of the WCM reduced the turnover rate of DA in the frontal cortex. The LWCM also attenuated the elevation of serum IL-1β induced by chronic stress. All doses of the WCM attenuated the ROCK2 protein hyperactivation, and the LWCM further increased the down-regulation of p-Akt/Akt signaling.
The WCM has antidepressant-like effects, which may result from the regulation of the stress-related ROCK2/PTEN/Akt pathway. Therefore, the WCM may be developed and used for the complementary treatment of depression.
ROCK-2-selective targeting and its therapeutic outcomes
2020, Drug Discovery Today
Citation Excerpt :
A negative feedback loop has also been identified in which the phosphorylated RhoE GTPase is involved in the inactivation of ROCK-1 but not of ROCK-2 [9]. ROCK inactivation and/or inhibition promotes several biological processes, including bone formation [30], neuronal morphogenesis [31,32], and keratinocyte differentiation [33], while also attenuating hypoxia-induced angiogenesis [34]. These functions, mediated via ROCK inhibition, are significant and form the basis for the development of ROCK inhibitors.
Despite the identification of distinct isoforms of Rho-kinase (ROCK-1 and ROCK-2), their isoform-specific roles in several disorders remain obscure. Recent studies have revealed a vital role of ROCK-2 in various vascular and neuronal disorders, where the potential for disease alleviation is wider with ROCK-2-selective targeting than with nonspecific ROCK inhibition. This approach is also crucial for resolving issues of safety and specificity associated with nonspecific ROCK inhibitors. In this review, we focus on the latest developments concerning ROCK-2 as a therapeutic target and justify the clinical use of ROCK-2-selective inhibitors.

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^☆: Abbreviations used: NGS, normal goat serum; Rho-kinase, Rho-associated kinase; NHE1, Na-H exchanger 1; NMDA, N-methyl d-aspartic acid; PAGE, polyacrylamide gel electrophoresis; PKN, protein kinase N; PSD, postsynaptic density; SDS, sodium dodecyl sulfate; TBS, Tris–HCl-buffered saline

¹: To whom correspondence should be addressed. Fax: +81-6-6879-3059. E-mail: [email protected].

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Regular ArticleDistribution of Rho-Kinase in the Bovine Brain☆

Abstract

Trends Biochem. Sci.

FEBS Lett.

J. Biol. Chem.

FEBS Lett.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Brain Res. Mol. Brain Res.

Brain Res. Mol. Brain Res.

Brain Res.

Science

Regular Article
Distribution of Rho-Kinase in the Bovine Brain☆