Endogenous Neurosteroid (3α,5α)3-Hydroxypregnan-20-one Inhibits Toll-like-4 Receptor Activation and Pro-inflammatory Signaling in Macrophages and Brain

Scientific Reports

Volumn 9, Issue 1, 2019, Pages

  Balan, Irina ^a   Beattie, Matthew C ^b   O’Buckley, Todd K ^b   Aurelian, Laure ^a   Morrow, A Leslie ^b  

^a UNIVERSITY OF MARYLAND SCHOOL OF MEDICINE   (United States)

^b UNIVERSITY OF NORTH CAROLINA SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ELTANOLONE; LIPOPOLYSACCHARIDE; PREGNENOLONE; TLR4 PROTEIN, MOUSE; TLR4 PROTEIN, RAT; TOLL LIKE RECEPTOR 4;

ANIMAL; IMMUNOLOGY; INFLAMMATION; MALE; METABOLISM; MOUSE; PROTEIN ANALYSIS; RAT; RAW 264.7 CELL LINE; SIGNAL TRANSDUCTION; VENTRAL TEGMENTUM;

ANIMALS; INFLAMMATION; LIPOPOLYSACCHARIDES; MALE; MICE; NEUROSTEROIDS; PREGNANOLONE; PREGNENOLONE; PROTEIN INTERACTION MAPS; RATS; RAW 264.7 CELLS; SIGNAL TRANSDUCTION; TOLL-LIKE RECEPTOR 4; VENTRAL TEGMENTAL AREA;

EID: 85061052580     PISSN: None     EISSN: 20452322     Source Type: Journal    
DOI: 10.1038/s41598-018-37409-6     Document Type: Article

References (100)

1. Selye, H. The anesthetic effect of steroid hormones. Proc Soc Exp Biol Med 46, 116–121 (1941)
2. McEwen, B. S. Non-genomic and genomic effects of steroids on neural activity. Trends Pharmacol Sci 12, 141–147 (1991)
3. Majewska, M. D., Harrison, N. L., Schwartz, R. D., Barker, J. L. & Paul, S. M. Steroid hormone metabolites are barbiturate-like modulators of the GABA receptor. Science 232, 1004–1007 (1986)
4. Morrow, A. L., Suzdak, P. D. & Paul, S. M. Steroid hormone metabolites potentiate GABA receptor-mediated chloride ion flux with nanomolar potency. Eur J Pharmacol 142, 483–485 (1987)
5. A receptors. Proc Natl Acad Sci USA 100, 14439–14444 (2003)
6. Harrison, N. L., Majewska, M. D., Harrington, J. W. & Barker, J. L. Structure-activity relationships for steroid interaction with the gamma-aminobutyric acid-A receptor complex. J Pharmacol Exp Ther 241, 346–353 (1987)
7. Paul, S. M. & Purdy, R. H. Neuroactive steroids. FASEB Journal 6, 2311–2322 (1992)
8. Patchev, V. K., Shoaib, M., Holsboer, F. & Almeida, O. F. X. The neurosteroid tetrahydroprogesterone counteracts corticotropin-releasing hormone-induced anxiety and alters the release and gene expression of corticotropin-releasing hormone in the rat hypothalamus. Neuroscience 62, 265–271 (1994)
9. Owens, M. J., Ritchie, J. C. & Nemeroff, C. B. 5α-Pregnane-3α,21-diol-20-one (THDOC) attenuates mild stress-induced increases in plasma corticosterone via a non-glucocorticoid mechanism: comparison with alprazolam. Brain Res 573, 353–355 (1992)
10. Cook, J. B. et al. Overexpression of the steroidogenic enzyme cytochrome P450 side chain cleavage in the ventral tegmental area increases 3alpha,5alpha-THP and reduces long-term operant ethanol self-administration. J Neurosci 34, 5824–5834, 10.1523/JNEUROSCI.4733-13.2014 (2014)
11. Beattie, M. C. et al. Voluntary ethanol consumption reduces GABAergic neuroactive steroid (3α,5α)3-hydroxypregnan-20-one (3α,5α-THP) in the amygdala of the cynomolgus monkey. Addict Biol 22, 318–330, 10.1111/adb.12326 (2017)
12. He, J., Hoffman, S. W. & Stein, D. G. Allopregnanolone, a progesterone metabolite, enhances behavioral recovery and decreases neuronal loss after traumatic brain injury. Restor Neurol Neurosci 22, 19–31 (2004)
13. Noorbakhsh, F., Baker, G. B. & Power, C. Allopregnanolone and neuroinflammation: a focus on multiple sclerosis. Front Cell Neurosci 8, 134, 10.3389/fncel.2014.00134 (2014)
14. Schumacher, M., Guennoun, R., Stein, D. G. & De Nicola, A. F. Progesterone: therapeutic opportunities for neuroprotection and myelin repair. Pharmacol Ther 116, 77–106 (2007)
15. Irwin, R. W. & Brinton, R. D. Allopregnanolone as regenerative therapeutic for Alzheimer’s disease: Translational development and clinical promise. Prog Neurobiol, doi:S0301-0082(13)00082-8, https://doi.org/10.1016/j.pneurobio.2013.08.004 (2013)
16. Wright, D. W. et al. ProTECT: a randomized clinical trial of progesterone for acute traumatic brain injury. Ann Emerg Med 49, 391–402 (2007)
17. Marx, C. E. et al. Proof-of-concept trial with the neurosteroid pregnenolone targeting neurocognitive and negative symptoms in schizophrenia. Biol Psychiatry 61, 13S (2007)
18. Fox, H. C., Sofuoglu, M., Morgan, P. T., Tuit, K. L. & Sinha, R. The effects of exogenous progesterone on drug craving and stress arousal in cocaine dependence: impact of gender and cue type. Psychoneuroendocrinology 38, 1532–1544, 10.1016/j.psyneuen.2012.12.022 (2013)
19. Milivojevic, V., Fox, H. C., Sofuoglu, M., Covault, J. & Sinha, R. Effects of progesterone stimulated allopregnanolone on craving and stress response in cocaine dependent men and women. Psychoneuroendocrinology 65, 44–53, 10.1016/j.psyneuen.2015.12.008 (2016)
20. Kanes, S. et al. Brexanolone (SAGE-547 injection) in post-partum depression: a randomised controlled trial. Lancet 390, 480–489, 10.1016/S0140-6736(17)31264-3 (2017)
21. Pavlov, V. A. & Tracey, K. J. Neural regulation of immunity: molecular mechanisms and clinical translation. Nat Neurosci 20, 156–166, 10.1038/nn.4477 (2017)
22. Walter, T. J., Vetreno, R. P. & Crews, F. T. Alcohol and Stress Activation of Microglia and Neurons: Brain Regional Effects. Alcohol Clin Exp Res, 10.1111/acer.13511 (2017)
23. He, J., Evans, C. O., Hoffman, S. W., Oyesiku, N. M. & Stein, D. G. Progesterone and allopregnanolone reduce inflammatory cytokines after traumatic brain injury. Exp Neurol 189, 404–412 (2004)
24. Ahmad, A. et al. Absence of TLR4 reduces neurovascular unit and secondary inflammatory process after traumatic brain injury in mice. PLoS One 8, e57208, 10.1371/journal.pone.0057208 (2013)
25. He, J. & Crews, F. T. Increased MCP-1 and microglia in various regions of the human alcoholic brain. Exp Neurol 210, 349–358, 10.1016/j.expneurol.2007.11.017 (2008)
26. Qin, L. et al. Increased systemic and brain cytokine production and neuroinflammation by endotoxin following ethanol treatment. J Neuroinflammation 5, 10, 10.1186/1742-2094-5-10 (2008)
27. Lacagnina, M. J., Rivera, P. D. & Bilbo, S. D. Glial and Neuroimmune Mechanisms as Critical Modulators of Drug Use and Abuse. Neuropsychopharmacology 42, 156–177, 10.1038/npp.2016.121 (2017)
28. Bhattacharya, A., Derecki, N. C., Lovenberg, T. W. & Drevets, W. C. Role of neuro-immunological factors in the pathophysiology of mood disorders. Psychopharmacology (Berl) 233, 1623–1636, 10.1007/s00213-016-4214-0 (2016)
29. Dantzer, R., O’Connor, J. C., Freund, G. G., Johnson, R. W. & Kelley, K. W. From inflammation to sickness and depression: when the immune system subjugates the brain. Nat Rev Neurosci 9, 46–56, 10.1038/nrn2297 (2008)
30. Maroso, M. et al. Interleukin-1 type 1 receptor/Toll-like receptor signalling in epilepsy: the importance of IL-1beta and high-mobility group box 1. J Intern Med 270, 319–326, 10.1111/j.1365-2796.2011.02431.x (2011)
31. Irie, T., Muta, T. & Takeshige, K. TAK1 mediates an activation signal from toll-like receptor(s) to nuclear factor-kappaB in lipopolysaccharide-stimulated macrophages. FEBS Lett 467, 160–164 (2000)
32. Lu, Y. C., Yeh, W. C. & Ohashi, P. S. LPS/TLR4 signal transduction pathway. Cytokine 42, 145–151, 10.1016/j.cyto.2008.01.006 (2008)
33. Chattopadhyay, S. & Sen, G. C. Tyrosine phosphorylation in Toll-like receptor signaling. Cytokine Growth Factor Rev 25, 533–541, 10.1016/j.cytogfr.2014.06.002 (2014)
34. Cochet, F. & Peri, F. The Role of Carbohydrates in the Lipopolysaccharide (LPS)/Toll-Like Receptor 4 (TLR4) Signalling. Int J Mol Sci 18, https://doi.org/10.3390/ijms18112318 (2017)
35. Kim, S. J. & Kim, H. M. Dynamic lipopolysaccharide transfer cascade to TLR4/MD2 complex via LBP and CD14. BMB Rep 50, 55–57 (2017)
36. Samad, T. A. et al. Interleukin-1beta-mediated induction of Cox-2 in the CNS contributes to inflammatory pain hypersensitivity. Nature 410, 471–475, 10.1038/35068566 (2001)
37. Thomson, C. A., McColl, A., Cavanagh, J. & Graham, G. J. Peripheral inflammation is associated with remote global gene expression changes in the brain. J Neuroinflammation 11, 73, 10.1186/1742-2094-11-73 (2014)
38. Fu, H. Q. et al. Prolonged neuroinflammation after lipopolysaccharide exposure in aged rats. PLoS One 9, e106331, 10.1371/journal.pone.0106331 (2014)
39. Crews, F. T., Lawrimore, C. J., Walter, T. J. & Coleman, L. G. Jr. The role of neuroimmune signaling in alcoholism. Neuropharmacology 122, 56–73, 10.1016/j.neuropharm.2017.01.031 (2017)
40. Okun, E., Griffioen, K. J. & Mattson, M. P. Toll-like receptor signaling in neural plasticity and disease. Trends Neurosci 34, 269–281, 10.1016/j.tins.2011.02.005 (2011)
41. June, H. L. et al. CRF-amplified neuronal TLR4/MCP-1 signaling regulates alcohol self-administration. Neuropsychopharmacology 40, 1549–1559, 10.1038/npp.2015.4 (2015)
42. Aurelian, L., Warnock, K. T., Balan, I., Puche, A. & June, H. TLR4 signaling in VTA dopaminergic neurons regulates impulsivity through tyrosine hydroxylase modulation. Transl Psychiatry 6, e815, 10.1038/tp.2016.72 (2016)
43. Balan, I., Warnock, K. T., Puche, A. & Gondre-Lewis, M. C. & Aurelian, L. Innately activated TLR4 signal in the nucleus accumbens is sustained by CRF amplification loop and regulates impulsivity. Brain Behav Immun 69, 139–153, 10.1016/j.bbi.2017.11.008 (2018)
44. Maroso, M. et al. Interleukin-1beta biosynthesis inhibition reduces acute seizures and drug resistant chronic epileptic activity in mice. Neurotherapeutics 8, 304–315, 10.1007/s13311-011-0039-z (2011)
45. Liu, J. et al. Binge alcohol drinking is associated with GABAA alpha2-regulated Toll-like receptor 4 (TLR4) expression in the central amygdala. Proc Natl Acad Sci USA 108, 4465–4470, 10.1073/pnas.1019020108 (2011)
46. Tsatsanis, C. et al. Corticotropin-releasing factor and the urocortins induce the expression of TLR4 in macrophages via activation of the transcription factors PU.1 and AP-1. J Immunol 176, 1869–1877 (2006)
47. Whitman, B. A., Knapp, D. J., Werner, D. F., Crews, F. T. & Breese, G. R. The cytokine mRNA increase induced by withdrawal from chronic ethanol in the sterile environment of brain is mediated by CRF and HMGB1 release. Alcohol Clin Exp Res 37, 2086–2097, 10.1111/acer.12189 (2013)
48. Lowery-Gionta, E. G. et al. Corticotropin releasing factor signaling in the central amygdala is recruited during binge-like ethanol consumption in C57BL/6J mice. J Neurosci 32, 3405–3413, 10.1523/JNEUROSCI.6256-11.2012 (2012)
49. Koob, G. F. et al. Addiction as a stress surfeit disorder. Neuropharmacology 76(Pt B), 370–382, 10.1016/j.neuropharm.2013.05.024 (2014)
50. Phillips, T. J., Reed, C. & Pastor, R. Preclinical evidence implicating corticotropin-releasing factor signaling in ethanol consumption and neuroadaptation. Genes Brain Behav 14, 98–135, 10.1111/gbb.12189 (2015)
51. Gondre-Lewis, M. C. et al. Early life stress is a risk factor for excessive alcohol drinking and impulsivity in adults and is mediated via a CRF/GABA(A) mechanism. Stress 19, 235–247, 10.3109/10253890.2016.1160280 (2016)
52. Dedic, N., Chen, A. & Deussing, J. M. The CRF family of neuropeptides and their receptors - mediators of the central stress response. Curr Mol Pharmacol, 10.2174/1874467210666170302104053 (2017)
53. Purdy, R. H., Morrow, A. L., Blinn, J. R. & Paul, S. M. Synthesis, metabolism, and pharmacological activity of 3a-hydroxy steroids which potentiate GABA-receptor-mediated chloride ion uptake in rat cerebral cortical synaptoneurosomes. J Med Chem 33, 1572–1581 (1990)
54. Patchev, V. K., Holsboer, F. & Almeida, O. F. X. In The Brain: Source and Target for Sex Steroid Hormones. (eds A. R. Genazzani, F. Petraglia, & R. H. Purdy) 93–102 (The Parthenon Group, 1996)
55. Scaffidi, P., Misteli, T. & Bianchi, M. E. Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature 418, 191–195, 10.1038/nature00858 (2002)
56. Andersson, U. & Tracey, K. J. HMGB1 is a therapeutic target for sterile inflammation and infection. Annu Rev Immunol 29, 139–162, 10.1146/annurev-immunol-030409-101323 (2011)
57. Morita, N. et al. C4b-binding protein negatively regulates TLR4/MD-2 response but not TLR3 response. FEBS Lett 591, 1732–1741, 10.1002/1873-3468.12693 (2017)
58. Chandrashekaran, I. R., Norton, R. S. & Schmitz-Peiffer, C. Characterisation of peptide interactions that regulate PKCepsilon activation. FEBS Lett 592, 179–189, 10.1002/1873-3468.12953 (2018)
59. Faraz, M., Herdenberg, C., Holmlund, C., Henriksson, R. & Hedman, H. A protein interaction network centered on leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) regulates growth factor receptors. J Biol Chem 293, 3421–3435, 10.1074/jbc.M117.807487 (2018)
60. Yang, H. et al. A critical cysteine is required for HMGB1 binding to Toll-like receptor 4 and activation of macrophage cytokine release. Proc Natl Acad Sci USA 107, 11942–11947, 10.1073/pnas.1003893107 (2010)
61. Quadros, I. M., Macedo, G. C., Domingues, L. P. & Favoretto, C. A. An Update on CRF Mechanisms Underlying Alcohol Use Disorders and Dependence. Front Endocrinol (Lausanne) 7, 134, 10.3389/fendo.2016.00134 (2016)
62. Park, J. S. et al. Involvement of toll-like receptors 2 and 4 in cellular activation by high mobility group box 1 protein. J Biol Chem 279, 7370–7377, 10.1074/jbc.M306793200 (2004)
63. Silva, E. et al. HMGB1 and LPS induce distinct patterns of gene expression and activation in neutrophils from patients with sepsis-induced acute lung injury. Intensive Care Med 33, 1829–1839, 10.1007/s00134-007-0748-2 (2007)
64. Balan, I. et al. The GABAA Receptor alpha2 Subunit Activates a Neuronal TLR4 Signal in the Ventral Tegmental Area that Regulates Alcohol and Nicotine Abuse. Brain Sci 8, 10.3390/brainsci8040072 (2018)
65. Patchev, V. K., Hassan, A. H. S., Holsboer, F. & Almeida, O. F. X. The neurosteroid tetrahydroprogesterone attenuates the endocrine response to stress and exerts glucocorticoid-like effects on vasopressin gene transcription in the rat hypothalamus. Neuropsychopharmacology 15, 533–540 (1996)
66. Morrow, A. L. Recent developments in the significance and therapeutic relevance of neuroactive steroids - Introduction to the special issue. Pharmacol Ther 116, 1–6 (2007)
67. Lawrimore, C. J. & Crews, F. T. Ethanol, TLR3, and TLR4 Agonists Have Unique Innate Immune Responses in Neuron-Like SH-SY5Y and Microglia-Like BV2. Alcohol Clin Exp Res 41, 939–954, 10.1111/acer.13368 (2017)
68. Sayeed, I., Guo, Q., Hoffman, S. W. & Stein, D. G. Allopregnanolone, a progesterone metabolite, is more effective than progesterone in reducing cortical infarct volume after transient middle cerebral artery occlusion. Ann Emerg Med 47, 381–389, 10.1016/j.annemergmed.2005.12.011 (2006)
69. Lehmann, S. M. et al. An unconventional role for miRNA: let-7 activates Toll-like receptor 7 and causes neurodegeneration. Nat Neurosci 15, 827–835, 10.1038/nn.3113 (2012)
70. Bsibsi, M. et al. The microtubule regulator stathmin is an endogenous protein agonist for TLR3. J Immunol 184, 6929–6937, 10.4049/jimmunol.0902419 (2010)
71. Kokate, T. G., Cohen, A. L., Karp, E. & Rogawski, M. A. Neuroactive steroids protect against pilocarpine- and kainic acid induced-limbic seizures and status epilepticus in mice. Neuropharmacology 35, 1049–1056 (1996)
72. Devaud, L. L., Purdy, R. H. & Morrow, A. L. The neurosteroid, 3α-hydroxy-5α-pregnan-20-one, protects against bicuculline-induced seizures during ethanol withdrawal in rats. Alcohol Clin Exp Res 19, 350–355 (1995)
73. Morrow, A. L., VanDoren, M. J., Penland, S. N. & Matthews, D. B. The role of GABAergic neuroactive steroids in ethanol action, tolerance and dependence. Brain Res Brain Res Rev 37, 98–109 (2001)
74. Porcu, P. & Morrow, A. L. Divergent neuroactive steroid responses to stress and ethanol in rat and mouse strains: relevance for human studies. Psychopharmacology (Berl) 231, 3257–3272, 10.1007/s00213-014-3564-8 (2014)
75. Marx, C. E. et al. Proof-of-concept trial with the neurosteroid pregnenolone targeting cognitive and negative symptoms in schizophrenia. Neuropsychopharmacology 34, 1885–1903, 10.1038/npp.2009.26 (2009)
76. Brinton, R. D. & Wang, J. M. Preclinical analyses of the therapeutic potential of allopregnanolone to promote neurogenesis in vitro and in vivo in transgenic mouse model of Alzheimer’s disease. Curr Alzheimer Res 3, 11–17 (2006)
77. Mayfield, J. & Harris, R. A. The Neuroimmune Basis of Excessive Alcohol Consumption. Neuropsychopharmacology 42, 376, 10.1038/npp.2016.177 (2017)
78. Pascual, M. et al. Gender differences in the inflammatory cytokine and chemokine profiles induced by binge ethanol drinking in adolescence. Addict Biol 22, 1829–1841, 10.1111/adb.12461 (2017)
79. Alfonso-Loeches, S., Pascual-Lucas, M., Blanco, A. M., Sanchez-Vera, I. & Guerri, C. Pivotal role of TLR4 receptors in alcohol-induced neuroinflammation and brain damage. J Neurosci 30, 8285–8295, 10.1523/JNEUROSCI.0976-10.2010 (2010)
80. Crews, F. T., Zou, J. & Qin, L. Induction of innate immune genes in brain create the neurobiology of addiction. Brain Behav Immun 25(Suppl 1), S4–S12, 10.1016/j.bbi.2011.03.003 (2011)
81. Pascual, M., Balino, P., Alfonso-Loeches, S., Aragon, C. M. & Guerri, C. Impact of TLR4 on behavioral and cognitive dysfunctions associated with alcohol-induced neuroinflammatory damage. Brain Behav Immun 25(Suppl 1), S80–91, 10.1016/j.bbi.2011.02.012 (2011)
82. Blednov, Y. A. et al. Activation of inflammatory signaling by lipopolysaccharide produces a prolonged increase of voluntary alcohol intake in mice. Brain Behav Immun 25(Suppl 1), S92–S105, 10.1016/j.bbi.2011.01.008 (2011)
83. Pandey, S. C. TLR4-MyD88 signalling: a molecular target for alcohol actions. Br J Pharmacol 165, 1316–1318, 10.1111/j.1476-5381.2011.01695.x (2012)
84. Leclercq, S. et al. Role of intestinal permeability and inflammation in the biological and behavioral control of alcohol-dependent subjects. Brain Behav Immun 26, 911–918, 10.1016/j.bbi.2012.04.001 (2012)
85. Alfonso-Loeches, S., Urena-Peralta, J., Morillo-Bargues, M. J., Gomez-Pinedo, U. & Guerri, C. Ethanol-Induced TLR4/NLRP3 Neuroinflammatory Response in Microglial Cells Promotes Leukocyte Infiltration Across the BBB. Neurochem Res 41, 193–209, 10.1007/s11064-015-1760-5 (2016)
86. Crews, F. T., Walter, T. J., Coleman, L. G. Jr. & Vetreno, R. P. Toll-like receptor signaling and stages of addiction. Psychopharmacology (Berl) 234, 1483–1498, 10.1007/s00213-017-4560-6 (2017)
87. Sanada, T. et al. FLN29 deficiency reveals its negative regulatory role in the Toll-like receptor (TLR) and retinoic acid-inducible gene I (RIG-I)-like helicase signaling pathway. J Biol Chem 283, 33858–33864, 10.1074/jbc.M806923200 (2008)
88. Vartanian, K. & Stenzel-Poore, M. Toll-like receptor tolerance as a mechanism for neuroprotection. Transl Stroke Res 1, 252–260, 10.1007/s12975-010-0033-5 (2010)
89. Laing, J. M., Smith, C. C. & Aurelian, L. Multi-targeted neuroprotection by the HSV-2 gene ICP10PK includes robust bystander activity through PI3-K/Akt and/or MEK/ERK-dependent neuronal release of vascular endothelial growth factor and fractalkine. J Neurochem 112, 662–676, 10.1111/j.1471-4159.2009.06475.x (2010)
90. Winkler, Z. et al. Impaired microglia fractalkine signaling affects stress reaction and coping style in mice. Behav Brain Res 334, 119–128, 10.1016/j.bbr.2017.07.023 (2017)
91. Sawle, P. et al. Carbon monoxide-releasing molecules (CO-RMs) attenuate the inflammatory response elicited by lipopolysaccharide in RAW264.7 murine macrophages. Br J Pharmacol 145, 800–810, 10.1038/sj.bjp.0706241 (2005)
92. Lee, J. W. et al. Characterization of the changes in eicosanoid profiles of activated macrophages treated with 20(S)-ginsenoside Rg3. J Chromatogr B Analyt Technol Biomed Life Sci 1065-1066, 14–19, 10.1016/j.jchromb.2017.09.002 (2017)
93. A alpha1 receptor function and trafficking in cultured cerebral cortical neurons. J Pharmacol Exp Ther 345, 317–325, 10.1124/jpet.112.201954 (2013)
94. Poulopoulos, A. et al. Neuroligin 2 drives postsynaptic assembly at perisomatic inhibitory synapses through gephyrin and collybistin. Neuron 63, 628–642, 10.1016/j.neuron.2009.08.023 (2009)
95. Smith, C. C., Yu, Y. X., Kulka, M. & Aurelian, L. A novel human gene similar to the protein kinase (PK) coding domain of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) codes for a serine-threonine PK and is expressed in melanoma cells. J Biol Chem 275, 25690–25699, 10.1074/jbc.M002140200 (2000)
96. VanDoren, M. J. et al. Neuroactive steroid 3α-hydroxy-5α-pregnan-20-one modulates electrophysiological and behavioral actions of ethanol. J Neurosci 20, 1982–1989 (2000)
97. Porcu, P. et al. Differential effects of ethanol on serum GABAergic 3alpha,5alpha/3alpha,5beta neuroactive steroids in mice, rats, cynomolgus monkeys, and humans. Alcohol Clin Exp Res 34, 432–442, 10.1111/j.1530-0277.2009.01123.x (2010)
98. Crawley, J. N., Glowa, J. R., Majewska, M. D. & Paul, S. M. Anxiolytic activity of an endogenous adrenal steroid. Brain Res 398, 382–385 (1986)
99. Sato, S., Yanagawa, Y., Hiraide, S. & Iizuka, K. Cyclic AMP signaling enhances lipopolysaccharide sensitivity and interleukin-33 production in RAW264.7 macrophages. Microbiol Immunol 60, 382–389, 10.1111/1348-0421.12381 (2016)
100. A receptors is a substrate for palmitoylation by GODZ. J Neurosci 24, 5881–5891 (2004)