Neurotransmitters, Mood and the Perception of Stress, Part 2

Glutamate, GABA and Neurosteroid Activation

Glutamate (L-glutamic acid) and GABA (gamma-aminobutyric acid) are, respectively, the principal excitatory and inhibitory neurotransmitters in the CNS and play a significant role in HPA axis function and mood.¹ These two amino acid-based neurotransmitters account for over 50% of the synapses in the brain, while the monoamines (serotonin, norepinephrine and dopamine) account for only about 5%.²

Glutamate is required for synaptic transmission and plasticity, and learning and memory. However, abnormal function of the glutamatergic system can lead to neurotoxicity, and has been implicated in the pathophysiology of several disorders, including amyotrophic lateral sclerosis (ALS), epilepsy, Huntington’s disease, Alzheimer’s disease, schizophrenia, depression and anxiety disorders.³ Glutamate signaling from the prefrontal cortex and hippocampus, through activation of N-methyl-D-aspartate (NMDA) receptors on the parvocellular neurons of the paraventricular nucleus, increases HPA activation.⁴ Subjects with depression (MDD) and bipolar disorder have notably higher glutamate levels, perhaps linking glutamate excitotoxicity with depression-related HPA axis activation. It is not known why glutamine levels (plasma and brain) are elevated in depressed subjects, but the potential to modulate glutamate-signaling remains a promising area of research.

NMDA receptors are primarily responsible for the regulation of glutamate activity. Abnormalities in NMDA receptor expression and binding affinities have been identified in patients with mood disorders.⁵ Alterations in glutamate binding through the glycine binding site on the NMDA receptor were identified in the frontal cortex of age-matched, post-mortem and interval-matched suicide victims.⁶ NMDA antagonists have been found to have antidepressant activity, while chronic antidepressant administration, including monoaminergic-based and trycyclic antidepressants, regulate NMDA receptor function and expression.⁷ Omega-3 fatty acids, considered to be helpful in some subjects with depression, have been shown to affect glutaminergic neurotransmission within the hippocampus.⁸

Magnesium and Zinc Modulation
Magnesium deficiency causes glutamate receptor (N-methyl-D-aspartate) calcium channels to open, resulting in an increase in glutaminergic activity and neuronal dysfunction. Both zinc and magnesium appear to modulate glutamate to nontoxic levels, preventing damage to the hippocampus and amygdala.⁹

Studies in depressed subjects suggest a higher likelihood of low magnesium, and find some benefits associated with the use of supplemental magnesium, though more research is needed to make specific dose recommendations.¹⁰ The role of magnesium is important enough in all metabolic processes that clinicians should consider measuring magnesium levels in depressed subjects and those with suspected HPA axis abnormalities. Clinicians should also be aware that most multivitamin-mineral products contain very little magnesium, or contain poorly absorbable forms of magnesium, such as MgO.

Serum zinc is also notably lower in depressed subjects.¹¹ Zinc supplementation (usually 25 mg zinc/day) has been used in several clinical trials with depressed subjects (often with antidepressant drugs) with some benefits, even when zinc levels are not measurably low.¹²

GABA and GABAergic Activities
Balancing excitotoxic levels of glutamate is of vital importance in relieving depressive and anxiety disorders, as well as certain severe neurological conditions. GABA is the primary inhibitory neurotransmitter within the CNS, serving as an important counterpart to the excitatory activities of glutamate. More specifically, GABA has been identified as the dominant inhibitory neurotransmitter within the hypothalamic paraventricular nucleus (PVN) where it exerts significant inhibitory tone upon HPA axis function.¹³ Lower plasma GABA levels have been reported in depressed and bipolar patients, levels which persist even after treatment with antidepressants.¹ Hypoactive GABAergic transmission within the PVN is therefore unable to properly down-regulate the HPA axis.

Increasing GABAergic activity can be accomplished by several non-pharmacological means (benzodiazepines are potent modulators of the GABA-A receptor, while gabapentin is a structural analog of GABA). A wide variety of natural ingredients, particularly flavonoid compounds are being explored for their GABAergic activities.^{14, 15} Food sources of GABA are limited, but can be found in some fermented foods, such as kimchi, kefir, miso, sauerkraut, tempeh and some yogurts. Clinical studies using food-derived GABA with HPA axis-related outcomes have not been reported. PharmaGABA^®, a naturally-sourced form of GABA purified from a fermentation process using the bacterial Lactobacillus hilgardii, has been investigated in humans, specifically examining its anxiolytic and immune-supporting effects during stress exposure.

In a double-blind study using 13 healthy volunteers, electroencephalogram (EEG) readings were obtained after 100 mg of PharmaGABA^® were administered in distilled water.¹⁶ These results were compared to distilled water alone (placebo) and distilled water containing 200 mg of L-theanine on separate days (7 day intervals). EEG waves were recorded at 0, 30 and 60 minutes after each administration. PharmaGABA^® produced a statistically significant increase in alpha waves, as well as a significant decrease in beta waves, when compared to the placebo. A significantly higher alpha to beta brain wave ratio was also found, even when compared to L-theanine administration.

In a separate study, Abdou et al. examined the effects of PharmaGABA^® using eight healthy volunteers with a history of acrophobia. Salivary IgA levels were measured as a biomarker for stress and immune response. Each participant was randomly assigned to either an experimental or placebo group. The participants crossed an extended pedestrian bridge while saliva samples were collected before crossing and at the middle and end of the bridge. The placebo group was found to have a decrease in IgA levels (decreased immune response) while the PharmaGABA^® group showed significantly higher levels. While additional studies of a larger size would help to demonstrate the immune and anxiolytic effects of GABA supplementation, these studies do point to the potential benefits of PharmaGABA^® in HPA-related stress, worry and anxiety following one hour of administration.¹⁷

Neurosteroids
Some of the most potent modulators of the GABA-A receptor are steroid compounds known as neurosteroids.¹⁸ Neurosteroids include a wide-range of pregnenolone-derived compounds, including DHEA, DHEA-S, pregnenolone-S, allopregnanolone (ALLO), and allotetrahydrodeoxycorticosterone (ALLO-THDOC). As the name implies, neurosteroids are compounds that have receptor-mediated neuromodulatory activities, although some clearly have other functions as well (e.g., DHEA acts as a precursor to androgens). There is some dispute about where the majority of these compounds are derived. They can be made in the brain, but several can enter the brain from the circulation (presumably made in the adrenal gland). Little is known about the regulation of their synthesis.

ALLO, the major neurosteroid in the brain, and ALLO-THDOC modulate feelings of depression and anxiety, as well as HPA axis stress. They affect several different pathways, including the GABA, glutamate, progesterone, and dopamine pathways.^19,20 Stress-induced reduction in the levels of ALLO and ALLO-THDOC lowers the inhibitory GABAergic transmission, allowing the HPA axis to become overactive. This blunted neurosteroid signaling is considered to be a critical mechanism in the vicious cyclical pathology of recurrent depression. In several studies, both plasma and CSF ALLO concentrations have been found to be decreased in patients with MDD and anxiety.^21,22 In patients with depression, an inverse relationship between ALLO concentrations and the severity of depressive illness has been reported.²³ In animal studies, pretreatment of rats with ALLO, ALLO-THDOC or progesterone has been shown to attenuate stress-induced increases in plasma ACTH and cortisol.²⁴

The use of supplemental DHEA and pregnenolone is common in clinical practice today. Many clinicians believe that the function of these hormones (usually consumed orally or sublingually) is primarily within the adrenal glands or in peripheral target tissue. However, in the case of pregnenolone specifically, oral doses appear to have demonstrable effects on neurological function (i.e. as a neurosteroid). In other words, oral pregnenolone appears to function as a precursor for CNS pregnenolone-S, and perhaps other neurosteroids like ALLO or ALLO-THDOC. For instance, ALLO serum levels have been reported to triple two hours after oral administration of 400 mg pregnenolone.²⁵ DHEA is also capable of passing into the CNS from the blood, and may also contribute neurosteroid effects.²⁶

This post is a modified excerpt of Dr. Guilliams’s book, The Role of Stress and the HPA Axis in Chronic Disease Management.

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² Leonard BE. Psychopathology of depression. Drugs Today (Barc). 2007 Oct;43(10):705-16.

³ Sanacora G, Zarate CA, Krystal JH, Manjii HK. Targeting the glutamatergic system to develop novel, improved therapeutic for mood disorders. Nature 2008; 7:426-437.

⁴ Durand D, Pampillo M, Caruso C, Lasaga M. Role of metabotropic glutamate receptors in the control of neuroendocrine function. Neuropharmacology. 2008 Sep;55(4):577-83.

⁵ Ohgi Y, Futamura T, Hashimoto K. Glutamate Signaling in Synaptogenesis and NMDA Receptors as Potential Therapeutic Targets for Psychiatric Disorders. Curr Mol Med. 2015;15(3):206-21.

⁶ Nowak G, Ordway GA, Paul IA. Alterations in the N-methyl-D-aspartate (NMDA) receptor complex in the frontal cortex of suicide victims. Brain Res 1995; 675: 157-164.

⁷ Nowak G, Trulias R, Layer RT, et al. Adaptive changes in the N-methyl-D-aspartate receptor complex after chronic treatment with imipramine and 1-aminocyclopropanecarboxylic acid. J Pharmacol Exp Ther 1993; 265:1380-1386.

⁸ Hennebelle M, Champeil-Potokar G, Lavialle M, Vancassel S, Denis I. Omega-3 polyunsaturated fatty acids and chronic stress-induced modulations of glutamatergic neurotransmission in the hippocampus. Nutr Rev. 2014 Feb;72(2):99-112.

⁹ Prior PL, Galduroz JC. Glutaminergic hyperfunctioning during alcohol withdrawal syndrome: therapeutic perspective with zinc and magnesium. Med Hypothesis 2011; 77(3): 368-70.

¹⁰ Derom ML, Sayón-Orea C, Martínez-Ortega JM, Martínez-González MA. Magnesium and depression: a systematic review. Nutr Neurosci. 2013 Sep;16(5):191-206.

¹¹ Siwek M, Dudek D, Schlegel-Zawadzka M, et al. Serum zinc level in depressed patients during zinc supplementation of imipramine treatment. J Affect Disord. 2010 Nov;126(3):447-52.

¹² Nowak G. Zinc, future mono/adjunctive therapy for depression: Mechanisms of antidepressant action. Pharmacol Rep. 2015 Jun;67(3):659-662.

¹³ Cullinan WE, Ziegler DR, Herman JP. Functional role of local GABAergic influences on the HPA axis. Brain Struct Funct. 2008 Sep;213(1-2):63-72.

¹⁴ Hanrahan JR, Chebib M, Johnston GA. Flavonoid modulation of GABA(A) receptors. Br J Pharmacol. 2011 May;163(2):234-45.

¹⁵ Nilsson J, Sterner O. Modulation of GABA(A) receptors by natural products and the development of novel synthetic ligands for the benzodiazepine binding site. Curr Drug Targets. 2011 Oct;12(11):1674-88.

¹⁶ Abdou AM, Higashiguchi S, Horie K, et al. Relaxation and immunity enhancement effects of ?-aminobutyric acid (GABA) administration in humans. BioFactors 2006;26: 201-208.

¹⁷ Alramadhan E, Hanna MS, Hanna MS, Goldstein TA, Avila SM, Weeks BS. Dietary and botanical anxiolytics. Medical Science Monitor?: International Medical Journal of Experimental and Clinical Research. 2012;18(4):RA40-RA48.

¹⁸ Crowley SK, Girdler SS. Neurosteroid, GABAergic and hypothalamic pituitary adrenal (HPA) axis regulation: what is the current state of knowledge in humans? Psychopharmacology (Berl). 2014 Sep;231(17):3619-34.

¹⁹ Bali A, Jaggi AS. Multifunctional aspects of allopregnanolone in stress and related disorders. Prog Neuropsychopharmacol Biol Psychiatry. 2014 Jan 3;48:64-78.

²⁰ Eser D, Schüle C, Baghai TC, Romeo E, Rupprecht R. Neuroactive steroids in depression and anxiety disorders: clinical studies. Neuroendocrinology. 2006;84(4):244-54

²¹ Esder D, Romeo E, Baghai TC, di Michele F, et al. Neuroactive steroids as modulators of depression and anxiety. Neuroscience 2006; 138:1041-1048.

²² Strohle A, Romeo E, Hermann B, et al. Concentrations of 3alpha-reduced neuroactive steroids and their precursors in plasma of patients major depression and after clinical recovery. Biol Psychiatry 1999;45: 274-277.

²³ Nappi RE, Petraglia F, Luisi S, et al. Serum allopregnanolone in women with postpartum "blues". Obstet Gynecol 2001; 97:77-80.

²⁴ Owens MJ, Ritchie JC, Nemeroff CB. 5 alpha-pregane-3 alpha, 2-diol-20-one (THDOC) attenuates mild stress-induced increases in plasma corticosterone via a non-glucocorticoid mechanism: comparison with alprazolam. Brain Res 1992; 573:353-355.

²⁵ Sripada RK, Marx CE, King AP, et al. Allopregnanolone elevations following pregnenolone administration are associated with enhanced activation of emotion regulation neurocircuits. Biol Psychiatry. 2013 Jun 1;73(11):1045-53.

²⁶ Sripada RK1, Welsh RC, Marx CE, Liberzon I. The neurosteroids allopregnanolone and dehydroepiandrosterone modulate resting-state amygdala connectivity. Hum Brain Mapp. 2014 Jul;35(7):3249-61.

About Thomas G. Guilliams, PhD
Dr. Guilliams earned his doctorate from the Medical College of Wisconsin (Milwaukee) where he studied molecular immunology in the Microbiology Department. Since 1996, he has spent his time studying the mechanisms and actions of natural-based therapies and is an expert in the therapeutic uses of nutritional supplements. As the Vice President of Scientific Affairs for Ortho Molecular Products, he has developed a wide array of products and programs which allow clinicians to use nutritional supplements and lifestyle interventions as safe, evidence-based and effective tools for a variety of patients. Tom teaches at the University of Wisconsin-School of Pharmacy, where he holds an appointment as a Clinical Instructor; at the University of Minnesota School of Pharmacy and is a faculty member of the Fellowship in Anti-aging Regenerative and Functional Medicine. He lives outside of Stevens Point, Wisconsin with his wife and children.