Main Research Findings

1) Treatment with alpha-GPC resulted in increased hippocampal neurogenesis and neuroprotection against seizure-induced cell death and the development of cognitive impairments.

2) Alpha-GPC was shown to protect against the enhanced cognitive decline and cellular damage as a result of focal brain irradiation.

 

Selected Data

1) Previous research has found that treatment with alpha-GPC has been found to improve cognitive function and enhance memory. The research team of Lee et al. examined whether the therapeutic abilities of the compound have the potential to enhance cognition and reduce neuronal cell death when induced by epileptic seizure. For the purpose of this study, 8 week old male Sprague-Dawley rats were utilized. Each animal was caged individually and maintained under standard laboratory conditions with a 12 hour light/dark cycle before induction of seizures took place [1].

In order to induce a seizure the animals were intraperitoneally injected with 127 mg/kg of lithium chloride, followed by 2 mg/kg of scopolamine and 25 mg/kg of pilocarpine, 19 hours later in order to ensure status epilepticus. Status epilepticus was typically achieved 20-30 minutes after injection of pilocarpine. The animals were then placed in individual observation cages in order for the research team to record seizure symptoms characterized by stereotypical mouse and facial movements, blinking, salivation, forelimb clonus, stiffened hind limbs, rearing and sinking [1].

Two hours after the onset of status epilepticus the animals were treated with 10 mg/kg of diazepam; 2 mg/kg of diazepam was delivered until recurrent seizure activity stopped. Following seizure induction the test subjects were divided into 4 groups including: sham seizure induction treated with a vehicle or alpha-GPC and seizure-induced animals treated with a vehicle or alpha-GPC. 250 mg/kg of alpha-GPC were administered to the animals once daily, starting immediately for 1 week or 3 weeks. For the late treatment group, the compound was administered once daily for 3 weeks starting 3 weeks after seizure induction. Instead of the active compound, the control group was administered 0.9% normal saline in the same doses as the experimental group [1].

In order to test the ability of alpha-GPC to improve seizure-induced cognitive impairments, the animals underwent the Morris water maze test two weeks after the seizure. The animals were pretested 1 week prior to administration of alpha-GPC, and then retested following treatment to identify improvements in cognitive functioning. The water maze training pool was filled with opaque water and included a circular escape stage submerged under the water’s surface. The testing took place over 5 days and 4 trials were completed each day. The trail began by placing the animal at a random starting point and allowing them to swim for 120 seconds or until they found the escape platform. A camera and tracking system recorded the route and swim trajectories of the animals for further analysis [1].

Following the treatment protocol and Morris water maze testing the animals were euthanized at 1, 3, or 6 weeks post-seizure. The brains were dissected and fixed in a preservative solution for 1 hour followed by fixation overnight with 30% sucrose solution to act as a cryoprotectant. After 2 days the brains were frozen with powered dry ice, cut with cryostats into 30 um thick slices, and maintained in a storage solution for further histological examination to take place. In order to evaluate the neuroprotective effects of alpha-GPC the brain sections were immunohistochemically stained by NeuN, followed by incubation with monoclonal anti mouse-NeuN antibody, biotinylated anti-mouse IgG, and ABC compound diluted in the primary antiserum. Immune responses were visualized with 3,3-diaminobenzidine and samples were mounted on slides to observe immunoreactions [1].

The research team then worked to detect blood brain barrier disruptions, as well as the presence of immature neurons. First, to detect disruptions in the blood brain barrier the ABC immunoperoxidase process was used to detect IgG-like immunoreactivity after staining brain sections with anti-rat IgG. Next, the presence of immature neurons were detected by immunostaining brain sections with guinea pig anti-DCX antibody followed by 24 hours of incubation with the primary DCX antibody. The sections were washed and incubated again in biotinylated goat anti-guinea pig diluted with the primary antiserum in order for the researchers to examine and quantify the intensity of dendritic signaling [1].

Finally, the research team detected activation of choline acetyltransferase (ChAT) by homogenizing the collected hippocampal specimens in a RIPA buffer compound followed by 20 minutes of centrifugation. The resulting supernatant was stored for further examination while hippocampal protein content was measured and the proteins were diluted, separated, and transferred to a polyvinylidene difluoride membrane. The membranes were then incubated with the primary antibody overnight, washed, and incubated again for an hour with anti-rabbit IgG secondary antibody conjugated with horseradish peroxidase [1].

2) Radiation therapy is frequently used to manage brain tumors, however, acute and chronic brain injury are severe side effects associated with this form of treatment. That being said, the research team of Plangár et al examine the ability of alpha-GPC to protect against the cognitive side effects of radiation therapy. For the purpose of this study, 24 6-week old male Sprague Dawley rats were utilized. 3-4 animals were housed together in one cage and maintained under standard laboratory conditions with ad libitum access to food and water. Prior to irradiation and treatment with the nootropic, the animals were randomly allocated to a treatment group and several separate experiments were conducted to determine the dose-effect relationship between single doses of irradiation and the effect on the brain. The randomly allocated treatment groups included: sham-irradiated control group, a group treated only with alpha-GPC, a group that was irradiated, and a group that was irradiated and treated with alpha-GPC [2].

Irradiation took place by anesthetizing the test subject and placing them in a prone position and fixing the ears with pins to plan where the radiation would be targeted. Ultimately, the research team chose to use a 6 MeV lateral electron beam at a 100 cm distance from the source to the skin as this method of delivery limits the radiation delivery to the hippocampus and corpus callosum while sparing the cerebellum, skin, eyes, ears, and the frontal lobe. Each animal had a 20 mm Newton metal insert with six 10 mm apertures placed into an electron applicator to receive irradiation or be considered sham-irradiated. Following the procedure the animals were placed back into their original cages as the researchers weighed them weekly, performed skin checks, and thoroughly observed and described their behavior. 1 week before the irradiation took place, the animals were administered a vehicle compound or a 50 mg/kg doses of alpha-GPC dissolved in 0.5 ml of sterile saline. Treatment was administered via oral gavage every Monday, Wednesday, and Friday for 4 months [2].

In order to assess how treatment with alpha-GPC affected cognition, learning, and memory, the mice were subjected to the Morris water maze test. The testing apparatus was a large circular tank, 60 cm in depth and filled with opaque water up to 40 cm depth. The apparatus was divided into 4 quadrants and an escape platform was randomly placed in one quadrant underneath the surface of the water. Four consecutive trials were performed and ended after 120 seconds or when the animal located the escape platform. If the platform was not located in the allocated time they were guided there by the researchers and left for 15 seconds before being removed from the testing chamber. The test subjects completed the Morris water maze test once prior to irradiation, once in the third month of treatment, and once in the fourth month of treatment [2].

Following treatment and behavioral testing the rats were euthanized and the brains were dissected and fixed in 4% paraformaldehyde for 24 hours. The preserved brain tissue was sliced and each section was embedded with paraffin for further histological evaluation that used a semiquantitative method for scoring the level of necrosis, macrophage density, reactive gliosis, calcification, and demyelination. Each of these parameters were typically scored on a scale of 1 to 5 with 1 representing no detection of damage, and 5 representing detection of severe damage [2].

 

Discussion

1) The research team of Lee et al examined the ability of alpha-GPC to reduce cognitive deficits and neuron death following seizure induction. When administered for only 1 week, the nootropic was shown to not elicit any significant effects on neuron death or disruptions in the blood brain barrier. Immunohistochemistry performed using anti-NeuN revealed that seizure induction led to neuronal death in the hippocampus, CA3, CA21, and subiculum. When comparing the experimental groups to the control group, 1 week of early treatment with alpha-GPC did not result in any significant difference in the number of live hippocampal neurons, the disruption of the blood brain barrier, or neuroprotective effects [1].

Figure 1: Changes in the amount of live NeuN (+) cells located in CA1, CA3, the hilus, and the hippocampus after 1 week of treatment with alpha-GPC administered immediately following seizure induction.

Comparatively, when alpha-GPC was administered for 3 weeks there was a significant reduction in hippocampal neuronal death and disruption of the blood brain barrier. The changes were detected using immunohistochemistry that revealed neuronal death and IfG leakage in the hippocampus following seizure induction. 3 weeks of immediate administration of alpha-GPC led to decreased neuron death in the CA3 and hilus regions when compared to the group treated with a vehicle. Similar results were reported in terms of IgG leakage in the experimental group compared to the control group. These results reported by the research team indicated that 3 weeks of treatment with alpha-GPC immediately following induction protected the hippocampus of the test subjects from neuronal death and blood brain barrier disruption [1].

Figure 2: Changes in the amount of live NeuN (+) cells located in CA1, CA3, the hilus, and the hippocampus after 3 weeks of treatment with alpha-GPC administered immediately following seizure induction.

In addition to immediate treatment with the nootropic, the research team also assessed how treatment administered 3 weeks after induction of injury affects hippocampal neuron death and disruption of the blood brain barrier. 6 weeks after the induction of seizure the brain sections collected from each group of test subjects were stained with NeuN and IgG. In the seizure induced group treated with a vehicle, the amount of live neurons in the hippocampus decreased by 62%. In the seizure induced group treated with alpha-GPC, there was only a 25% decrease in the number of live neurons in the hippocampus. Overall, these findings suggested that neuronal death and blood brain barrier disruption were significantly decreased when alpha-GPC was administered for 3 weeks, beginning 3 weeks after a seizure was induced [1].

Figure 3: Changes in the amount of live NeuN (+) cells located in CA1, CA3, the hilus, and the hippocampus after 3 weeks of treatment with alpha-GPC administered 3 weeks after seizure induction occurred.

Treatment with alpha-GPC was also examined to determine the ability of the compound to improve cognitive functioning measured through Morris water maze testing. The test subjects completed the water maze at 3 weeks post-seizure, before injection of alpha-GPC, and at 5 weeks post-seizure, after injection of alpha-GPC. Prior to treatment the animals were not able to reach the escape platform within the 120 second time limit. However, after treatment the animals were able to successfully reach the escape platform, indicating improved performance and cognitive functioning [1].

Figure 4: Changes in performance on the Morris water maze test recorded at 3 weeks post-seizure, prior to treatment with alpha-GPC, and 5 weeks post-seizure, after treatment with alpha-GPC.

2) Researchers Plangár et al assessed the neuroprotective abilities of alpha-GPC in Sprague Dawley rats subjected to brain irradiation. First, the research team thought it was important to mention that the rats included in the experimental group experienced a deficit in body weight that persisted throughout the entire experiment. That being said, at the end of the study the average weight of the animals in the experimental group was 470.83 grams, compared to the average weight of the control group of 515 grams. The difference in body weights between the groups was not deemed to be significant by the researchers [2].

Acquisition and retention of spatial working memory was assessed by having the test subjects complete the Morris water maze test. Following irradiation, there were significant changes in performance time in both treated and untreated irradiated animals. However, in the animals that underwent irradiation and were treated with alpha-GPC changes in performance time significantly improved, as well as spatial working memory and learning abilities. The test subjects in the experimental control group began to experience symptoms of neurological deterioration 90 days after irradiation occurred. This was compared to the control group that began to experience symptoms of neurological deterioration 120 days after irradiation. Additionally, the symptoms in the control group were more severe than those in the experimental group treated with alpha-GPC [2].

In terms of latency to finding the escape platform there were significant memory impairments observed after 120 days in the irradiated animals. However, when treated with alpha-GPC the learning latency was raised to a similar level to the control group of animals that did not receive irradiation. Latency of finding the escape platform was also significantly reduced following administration of alpha-GPC to the irradiated animals. Irradiation also led to the development of severe cognitive deficits in the test subjects that were significantly relieved following treatment with alpha-GPC. Overall these findings suggest that the nootropic improved memory and learning while reducing cognitive deficits [2].

Figure 5: Changes in the latency of finding the escape platform during the Morris water maze testing across all of the included experimental treatment groups.

Following euthanization the brains were dissected and sliced into sections that were stained with hematoxylin and eosin in order to identify signs of necrosis caused by irradiation. In the control group of animals and the sham-irradiated group of animals there were no signs of necrosis, reactive astrogliosis, or any other various histopathological categories. As for the irradiated animals, the researchers examined the brain sections for signs of necrosis, reactive gliosis, calcification, increased macrophage density, and axon demyelination. The untreated group of irradiated animals experienced necrosis of the gray and white matter, increased demyelination, destruction of fibers, macrophage density, and calcification, as well as evidence of reactive astrogliosis [2].

In the groups treated with alpha-GPC there were significant decreases in the scores for macrophage density, calcification, demyelination, and reactive gliosis over the 120 experimental period. When comparing the irradiated groups that were not treated versus those that were treated with alpha-GPC the scoring for necrosis was 3.33 versus 2.33; the scoring for macrophage density was 2.83 versus 1.50; the scoring for reactive astrogliosis was 3.50 versus 1.83; the scoring for calcification was 3.0 versus 2.0; and the scoring for demyelination was 3.17 versus 2.17, respectively. Based on the reported findings the researchers deemed that there were significant changes in scoring between the treated and untreated irradiated group in macrophage density, reactive gliosis, calcificant, and demyelination. However, changes in the amount of necrosis present in the two groups were not considered significant. The findings reported in the study concluded that treatment with alpha-GPC was able to improve symptoms and side effects related to irradiation treatment [2].

Figure 6: Changes in the extent of necrosis present after 120 days across all experimental treatment groups.

 

Disclaimer

**LAB USE ONLY**
*This information is for educational purposes only and does not constitute medical advice. THE PRODUCTS DESCRIBED HEREIN ARE FOR RESEARCH USE ONLY. All clinical research must be conducted with oversight from the appropriate Institutional Review Board (IRB). All preclinical research must be conducted with oversight from the appropriate Institutional Animal Care and Use Committee (IACUC) following the guidelines of the Animal Welfare Act (AWA).

 

Citations

[1] Lee SH, Choi BY, Kim JH, et al. Late treatment with choline alfoscerate (l-alpha glycerylphosphorylcholine, α-GPC) increases hippocampal neurogenesis and provides protection against seizure-induced neuronal death and cognitive impairment. Brain Res. 2017;1654(Pt A):66-76. doi:10.1016/j.brainres.2016.10.011

[2] Plangár I, Szabó ER, Tőkés T, et al. Radio-neuroprotective effect of L-alpha-glycerylphosphorylcholine (GPC) in an experimental rat model. J Neurooncol. 2014;119(2):253-261. doi:10.1007/s11060-014-1489-z

 

 

What is Alpha-GPC?

Alpha-GPC (Alpha-Glycerophosphocholine) is a choline-containing compound that has been shown to have cognitive-promoting properties and is considered a nootropic. It’s a naturally occurring compound found both in the brain and in food sources.

Here’s a more detailed look at Alpha-GPC:

1. Role in the Brain: Alpha-GPC is a precursor to the neurotransmitter acetylcholine. Acetylcholine plays a vital role in cognitive functions such as memory, attention, and learning. By supplying the brain with additional choline, Alpha-GPC can potentially help support the synthesis and release of acetylcholine.
2. Therapeutic Use: In some countries, Alpha-GPC is prescribed to treat cognitive decline, like that seen in Alzheimer’s disease. The idea is that by providing the brain with more choline, it can help boost the levels of acetylcholine, which typically decline in neurodegenerative conditions.
3. Nootropic and Athletic Use: Some individuals use Alpha-GPC as a dietary supplement for its potential cognitive-enhancing properties. Additionally, there’s interest in its possible benefits for enhancing power output in athletes, though more research is needed in this area.
4. Dietary Sources: Alpha-GPC is found in red meat, organ tissues, and some other food sources, though in smaller amounts than what’s typically found in supplements.
5. Safety and Dosage: While Alpha-GPC is generally considered safe, as with any supplement, it’s essential to consult with a healthcare professional regarding its use, especially concerning dosage and potential drug interactions.

Remember, while Alpha-GPC shows promise in various applications, it’s crucial to approach its use with an evidence-based perspective and to consult with a healthcare professional when considering supplementation.

Why is It Called Alpha-GPC?

Alpha-GPC stands for “Alpha-Glycerophosphocholine.” It’s a naturally occurring choline compound found in the brain and is also a precursor to the neurotransmitter acetylcholine. Breaking down the name:

• “Alpha” refers to a specific chemical structure or arrangement in organic chemistry.
• “Glycerophosphocholine” refers to the molecule’s composition. Glycerophosphocholine is a molecule that contains glycerol, phosphate, and choline.

Alpha-GPC has been studied for its potential cognitive-enhancing properties and is often taken as a nootropic supplement. The name essentially provides a description of the chemical structure and nature of the compound.

What Are The Mechanisms and Effects of Alpha-GPC?

Alpha-glycerophosphocholine (Alpha-GPC) is a choline compound that contains a phospholipid. The main benefits of the compound are improvement in cognition, primarily memory and attention, and the ability to help combat Alzheimer’s disease. The compound breaks down into choline and glycerol-1-phosphate. Choline is a precursor for acetylcholine, a neurotransmitter that plays a large role in both the central and peripheral nervous system. Glycerol-1-phosphate is shown to support and protect cellular membranes.

Researchers have theorized that Alpha-GPC works by increasing the synthesization and the expression of acetylcholine in the brain. This is what allows it to evoke such drastic effects on memory, learning, and attention. Acetylcholine is often closely related to Alzheimer’s as the most common treatment for the disease is acetylcholinesterase inhibitors that promote increased levels of acetylcholine in the brain (https://examine.com/supplements/alpha-gpc/).

Evidence has shown that Alpha-GPC also promotes the secretion of Growth Hormone (GH) due to higher circulating levels of choline. In addition to the increased levels of GH, acetylcholine is responsible for skeletal muscle contraction, which indicates involvement in the improvement of physical performance. Various studies have administered Alpha-GPC to animals and found that administration of the nootropic led to a drastic improvement in various physical performance tests. These studies also emphasized that even a short supplementation period led to increased muscle mass and improved performance.

 

Alpha-GPC as a Byproduct of Wheat Fermentation

As it was mentioned above, Alpha-GPC has shown promise in treating Alzheimer’s disease and other forms of dementia due to the correlation between the compound and the neurotransmitter, acetylcholine. Since the 1990s animal-based studies have been conducted in order to determine how effective Alpha-GPC is against memory loss. The animals being tested were given either a placebo or active Alpha-GPC for 10 days. After the 10 days memory loss was induced through use of the drug, scopolamine. Overall the study concluded that administration of Alpha-GPC was capable of significantly reducing memory impairments and promoting general improvement to cognitive functions associated with Alzheimer’s and dementia (https://www.onnit.com/academy/alpha-gpc-benefits/).

Due to the potential Alpha-GPC has shown in treating Alzheimer’s and memory loss, researchers have begun to find a sustainable way of synthesizing the compound. Researchers Oyeneye et. Al examined how the fermentation of wheat could possibly produce sufficient levels of Alpha-GPC. Current methods of production include the hydrolysis of phosphocholine or the condensation of glycerol derivatives with the use of catalysts. The enzymatic method of phosphocholine hydrolysis is effective as it reduces the amount of excess chemical reagents being produced. Further purification of Alpha-GPC comes from the use of chromatography. While this sufficiently produces Alpha-GPC it is not a sustainable method of synthesization and incredibly costly to the laboratories and researchers looking to further their experiments.

The researchers found that AC Andrew was the type of wheat cultivar best suited to produce large amounts of Alpha-GPC due to its high starch content. Typically when fermentation occurs ethanol is produced until it reaches its peak at approximately 72 hours, however this study highlighted the fact that after ethanol production had stopped, Alpha-GPC continued to form without altering the fermentation process at all. That being said, when the goal of fermentation is specifically to produce Alpha-GPC, there are no excess levels of ethanol produced. These findings support the goal of synthesizing Alpha-GPC in an energy-efficient and low expense manner (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7271372/).

 

GPC Promotion of Longevity and Health in Roundworms

In addition to its benefits on cognition, researchers Liu et. Al examined how GPC could combat symptoms of aging in Caenorhabditis elegans, more commonly known as the roundworm. Since GPC levels tend to decrease while aging, the roundworms were given an initial dosage of 10 mM of GPC to see how the compound would affect whole lifespan, mean lifespan, and maximum lifespan. The dosage capped at 50 mM and led to a drastic increase in whole, mean, and maximum lifespan.

Furthermore, it was noted that motor ability declines from aging so the researchers measured the roundworms’ activity levels by examining body bending rate and pharyngeal pumping rate at different points throughout their lifespan. Following a 50 mM dose of GPC there was a drastic improvement in both of these rates. Specifically, GPC increased the rate of body bending in the mid-late life stage and increased the rate of pharyngeal pumping in the early-mid life stage. These results suggest that treatment with GPC helps to alleviate the symptoms of age-induced decline (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875989/).

The nootropics sold by Umbrella Labs are sold for laboratory research only. The description above is not medical advice and is for informative purposes only.

Alpha GPC is sold for laboratory research use only. Terms of sale apply. Not for human consumption, nor medical, veterinary, or household uses. Please familiarize yourself with our Terms & Conditions prior to ordering.