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Exploring The Role of Aquaporin 4 in Hydrocephalus

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In a study conducted recently by researchers in the Department of Neurosurgery at Shandong Provincial Qianfoshan Hospital and Shandong University in China, the researchers sought to direct attention to the important role that a protein plays in the pathology of hydrocephalus. The researchers maintain that understanding the benefits, roles, and functions that this protein plays in the brain can possibly serve as a great motivation for undertaking a research work that would enhance effective therapeutic treatment for hydrocephalus. The researchers are of the view that since cerebrospinal fluid (CSF) circulation is defective in hydrocephalus, a promising therapeutic approach would be one that is geared towards regulating water channels in the brain. Hydrocephalus is a neurological disorder that stems from abnormal build-up of cerebrospinal fluid in the brain.

The regulation of this fluid can be disrupted when there is an imbalance between its production and absorption. Thus, the build-up of the cerebrospinal fluid can produce various impairments in neuronal functioning. The current therapeutic method of treating hydrocephalus is the use of shunt system, which is the use of a catheter to remove the excess fluid build-up in the brain; which, however, has a high failure rate. What water channels are speculated culprits? The researchers examined the role of aquaporins in the pathology of hydrocephalus, particularly aquaporin 4, a subtype of the aquaporin family. Aquaporins are water channel proteins that seek to maintain water balance in the cell, and play a role in neural signal transduction (a chemical or physical signal transmitted through a cell that results in a change of the state or function of a cell), and also helps in cell migration. The most common type of aquaporin in the brain is aquaporin 4 (AQP4), which plays a significant role in the blood-brain barrier (BBB) and brain-cerebrospinal fluid border, as well as regulates the movement of water in the brain. The blood-brain barrier separates blood from the brain, which involves the mix of firmly connected endothelial cells and brain cells that prevent pathogens and other toxins from entering the blood circulation that can potentially harm the brain. The endothelial cells cover blood capillaries tightly, so that harmful substances do not penetrate into the blood.

Astrocytes (a type of brain cells) latch onto the endothelial cells that line the blood vessels, thus, preventing pathogens and toxins from penetrating through the endothelial cells and into the blood. AQP4 is at the end feet (terminal end) of astrocytes, where it receives signals that finetunes its expression. AQP4 is the main water channel in the central nervous system. It also plays an important role in the physiological and pathological water balance in the brain, and it is linked to cases of brain edema (caused by imbalance of water in the brain). Besides, AQP4 functions differently in different types of brain edema. The researchers’ speculation was informed by previous related studies that found that AQP4 levels were increased in subjects with hydrocephalus. However, it is not clear as to whether AQP4 has a protective role against hydrocephalus or a contributive role. Thus, the researchers of this study hypothesized that AQP4 does contribute to the pathology of hydrocephalus. Materials and Methods In order to investigate the role of AQP4 in hydrocephalus, the researchers used a male rat model by injecting autologous blood as a way of establishing the presence of hydrocephalus. The researchers injected the rat’s own blood in its brain’s lateral ventricles, causing a build-up of fluid and disrupting the flow of CSF. The experiment was bipartite. The first part of the experiment was to establish a hydrocephalus model by injecting autologous blood into the ventricles (located in the brain) of the rat. The second part of the experiment consisted of 3 groups (18 rats in each group): the AQP4 siRNA group, the negative control of siRNA group, and control group. In the AQP4 siRNA group, a chemical that inhibits the expression of AQP4 was injected into the ventricles, and thereafter, the rats received an injection of autologous blood in the ventricles. For the negative control of siRNA group, the rats received injection of autologous blood and a chemical that does not target any gene production (this served as a baseline to compare with the group in which the expression or production of AQP4 was inhibited). The rats in the control group were injected with normal saline.

The procedures and experimental treatment were carried out in a period of 3 days. Results The brain tissues of rats in each group were examined with the use of an MRI after the injection of the autologous blood. The MRI revealed an enlargement of the ventricles, which shows that hydrocephalus was induced. In order to examine the expression of the AQP4 protein at each experimental condition, researchers sought to use techniques such as Evans blue extravasation assay, HE staining, immunohistochemistry, immunofluorescence, and qRT-PCR. These techniques have similar functions of examining target protein levels. In the hydrocephalus model, there was an increase in AQP4 level. This was detected through qRT-PCR and immunohistochemistry. These methods examined AQP4 levels. There was an increased expression of AQP4 in the hydrocephalus group when compared with the control group. AQP4 siRNA decreased the AQP4 in brain tissues. Western blot analysis (a method that analyzes protein levels) results showed that AQP4 siRNA effectively decreased the protein levels of AQP4 when compared with the negative control siRNA group. In a nutshell, AQP4 siRNA was able to reduce AQP4 level. Silencing AQP4 worsens hydrocephalus. After AQP4 was silenced, hydrocephalus worsened and the lateral ventricles were significantly enlarged as compared to other experimental groups.

Connexin 43 level, which was detected by some of the aforementioned techniques showed significant increase when hydrocephalus was induced. When hydrocephalus was induced, connexin 43 level increased and reduced when AQP4 was silenced. Connexin 43 is also a protein channel like aquaporins. Connexin 43 is a protein located in the gap junction that allows for the electrical communication between neurons. It has a functional relationship with AQP4. It also helps to maintain homeostasis in the brain, just like AQP4, and responds to hydrocephalus in a way similar to AQP4. The blood-brain barrier was also damaged by hydrocephalus, and was worst when AQP4 was silenced. Implications of Study and Future Directions In this study, the researchers found that hydrocephalus increased AQP4 levels. Silencing AQP4 intensified hydrocephalus and debilitated the state of the blood-brain barrier. The researchers highlighted that these findings suggest that AQP4 has a protective role in hydrocephalus. The researchers also discussed findings as it relates to the functional roles of AQP4 in different structures, as well as how these functional roles are hindered by hydrocephalus. This provides a platform for prospective studies that will aim at investigating and experimenting on different drugs that will modulate AQP4 receptors as a way of treating hydrocephalus. Since the findings suggest that the inhibition of the production of AQP4 affects hydrocephalus adversely; therefore, a future study should aim at developing a type of drug that can target the overexpression (increase in production) of AQP4, in order to regulate the excess fluid buildup in hydrocephalus.

This endeavor will be a breakthrough in the pharmaceutical field. Conclusion In effort to explore the role of Aquaporin 4 in hydrocephalus, the journal article titled “Aquaporin 4 Silencing Aggravates Hydrocephalus Induced by Injection of Autologous Blood in Rats” was examined. The researchers used a hydrocephalus rat model to show that AQP4 serves a protective role against hydrocephalus. When they induced hydrocephalus in the rat model, AQP4 level was elevated, and when they silenced the expression of AQP4, they found that hydrocephalus was intensified in the rat model. The researchers highlighted that regulating or fine-tuning AQP4 water channels may be a fascinating therapy for the treatment of hydrocephalus. Understanding the mechanisms and role of AQP4 in hydrocephalus, particularly further in-depth study on its role on other cerebrospinal fluid pathology, will shed light on various treatment strategies that can aid to the development of drugs that will elevate AQP4 level as a way to alleviate hydrocephalus. The question here is, “Can this knowledge be used for working on humans?” If so, how can this research help pharmaceutical companies to manufacture drugs that can enhance effective treatment of this imbalance in humans?

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Exploring The Role Of Aquaporin 4 In Hydrocephalus. (2019, August 27). GradesFixer. Retrieved August 9, 2022, from
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