Gintonin influences the morphology and motility of adult brain neurons via LPA receptors

Do-Geun Kim; Hyeon-Joong Kim; Sun-Hye Choi; Sung Min Nam; Hyoung-Chun Kim; Hyewhon Rhim; Ik-Hyun Cho; Man Hee Rhee; Seung-Yeol Nah

doi:10.1016/j.jgr.2020.06.003

Gintonin influences the morphology and motility of adult brain neurons via LPA receptors

J Ginseng Res. 2021 May;45(3):401-407. doi: 10.1016/j.jgr.2020.06.003. Epub 2020 Jul 21.

Authors

Do-Geun Kim^{1

2}, Hyeon-Joong Kim³, Sun-Hye Choi³, Sung Min Nam³, Hyoung-Chun Kim⁴, Hyewhon Rhim⁵, Ik-Hyun Cho⁶, Man Hee Rhee⁷, Seung-Yeol Nah³

Affiliations

¹ Dementia Research Group, Korea Brain Research Institute, Cheomdanro 61, Daegu, Republic of Korea.
² Department of Anatomy, School of Medicine and Institute for Environmental Science, Wonkwang University, Iksan, Republic of Korea.
³ Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University, Seoul, Republic of Korea.
⁴ Neuropsychopharmacology and Toxicology Program, College of Pharmacy, Kangwon National University, Chunchon, Republic of Korea.
⁵ Center for Neuroscience, Korea Institute of Science and Technology, Seoul, Republic of Korea.
⁶ Department of Convergence Medical Science, Brain Korea 21 Plus Program, and Institute of Korean Medicine, College of Korean Medicine, Kyung Hee University, Seoul, Republic of Korea.
⁷ Laboratory of Veterinary Physiology & Cell Signaling, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.

Abstract

Background: Gintonin is an exogenous ginseng-derived G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. LPA induces in vitro morphological changes and migration through neuronal LPA1 receptor. Recently, we reported that systemic administration of gintonin increases blood-brain barrier (BBB) permeability via the paracellular pathway and its binding to brain neurons. However, little is known about the influences of gintonin on in vivo neuron morphology and migration in the brain.

Materials and methods: We examined the effects of gintonin on in vitro migration and morphology using primary hippocampal neural precursor cells (hNPC) and in vivo effects of gintonin on adult brain neurons using real time microscopic analysis and immunohistochemical analysis to observe the morphological and locational changes induced by gintonin treatment.

Results: We found that treating hNPCs with gintonin induced morphological changes with a cell rounding following cell aggregation and return to individual neurons with time relapses. However, the in vitro effects of gintonin on hNPCs were blocked by the LPA1/3 receptor antagonist, Ki16425, and Rho kinase inhibitor, Y27632. We also examined the in vivo effects of gintonin on the morphological changes and migration of neurons in adult mouse brains using anti-NeuN and -neurofilament H antibodies. We found that acute intravenous administration of gintonin induced morphological and migrational changes in brain neurons. Gintonin induced some migrations of neurons with shortened neurofilament H in the cortex. The in vivo effects of gintonin were also blocked by Ki16425.

Conclusion: The present report raises the possibility that gintonin could enter the brain and exert its influences on the migration and morphology of adult mouse brain neurons and possibly explains the therapeutic effects of neurological diseases behind the gintonin administration.

Keywords: Adult brain neuron; BBB, blood brain barrier; BSA, bovine serum albumin; DAPI, 4′,6-diamidino-2-phenylindole; DMEM, Dulbecco's modified Eagle's medium; DMSO, dimethyl sulfoxide; EGF, epidermal growth factor; FITC, fluorescein isothiocyanate; Gintonin; HBSS, Hanks' Balanced Salt Solution; LPA receptors; LPA, Lysophatidic Acid; MEM, Modified Eagle's medium; Morphology and migration; NECAB1, Neuronal calcium binding proteins 1; NFH, neurofilament H; OCT, optimum cutting temperature; PFA, paraformaldehyde; ROCK, Rho-associated protein kinase; bFGF, fibroblast growth factor; hNPC, hippocampal neural precursor cells.