See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/301685682 Post-mortem brains of alcoholics: changes in the glutamatergic, serotonergic, endocannabinoid and neuroactive steroid systems Thesis · April 2016 DOI: 10.13140/RG.2.1.3236.3929 CITATIONS READS 0 446 1 author: Olli Kärkkäinen University of Eastern Finland 52 PUBLICATIONS 323 CITATIONS SEE PROFILE Some of the authors of this publication are also working on these related projects: Biologically Active Synthetic Coumarins View project Metabolomics and alcohol use View project All content following this page was uploaded by Olli Kärkkäinen on 28 April 2016. The user has requested enhancement of the downloaded file. publications of the university of eastern finlanD Dissertations | olli kärkkäinenDissertations brains | alcoholics... of | post-mortem Dissertations in Health Sciences OLLI KÄRKKÄINEN Alcoholics are a heterogeneous group with a spectrum of health problems. Alcohol in- fluences the function of many of the brain’s messaging systems. This thesis investi- gated the post-mortem brain samples of late-onset Cloninger type 1 and early-onset type 2 alcoholics. Increased dehydroepian- drosterone and pregnenolone levels, and decreased serotonin transporter binding, were observed in the alcoholics when com- pared to controls. Moreover, only antisocial type 2 alcoholics had increased AMPA receptor binding in the anterior cingula- te cortex and only anxiety-prone type 1 alcoholics showed increased docosahexa- enoylethanolamide levels in the amygdala. n o 333 333 uef.fi publications of OLLI KÄRKKÄINEN the university of eastern finlanD Dissertations in Health Sciences post-mortem brains of alcoholics Changes in the Glutamatergic, Serotonergic, Endocannabinoid and Neuroactive Steroid Systems ISBN 978-952-61-2051-5 ISSN 1798-5706 KÄRKKÄINEN OLLI Post-Mortem Brains of Alcoholics Changes in the Glutamatergic, Serotonergic, Endocannabinoid and Neuroactive Steroid Systems To be presented by permission of the Faculty of Health Sciences, University of Eastern Finland for public examination in auditorium CA102, Canthia building, Kuopio, on Monday, April 18th 2016, at 12 noon Publications of the University of Eastern Finland Dissertations in Health Sciences Number 333 Pharmacology and Toxicology, School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland Kuopio 2016 Juvenus Print Tampere, 2016 Series Editors: Professor Veli-Matti Kosma, M.D., Ph.D. Institute of Clinical Medicine, Pathology Faculty of Health Sciences Professor Hannele Turunen, Ph.D. Department of Nursing Science Faculty of Health Sciences Professor Olli Gröhn, Ph.D. A.I. Virtanen Institute for Molecular Sciences Faculty of Health Sciences Professor Kai Kaarniranta, M.D., Ph.D. Institute of Clinical Medicine, Ophthalmology Faculty of Health Sciences Lecturer Veli-Pekka Ranta, Ph.D. (pharmacy) School of Pharmacy Faculty of Health Sciences Distributor: University of Eastern Finland Kuopio Campus Library P.O.Box 1627 FI-70211 Kuopio, Finland http://www.uef.fi/kirjasto ISBN (print): 978-952-61-2051-5 ISBN (pdf): 978-952-61-2052-2 ISSN (print): 1798-5706 ISSN (pdf): 1798-5714 ISSN-L: 1798-5706 III Author’s address: School of Pharmacy, Faculty of Health Sciences University of Eastern Finland KUOPIO FINLAND Supervisors: Professor Jari Tiihonen, Ph.D., M.D. Forensic Psychiatry, School of Medicine, Faculty of Health Sciences, University of Eastern Finland Niuvanniemi Hospital KUOPIO FINLAND Adjunct Professor Markus Storvik, Ph.D. Pharmacology and Toxicology, School of Pharmacy, Faculty of Health Sciences University of Eastern Finland KUOPIO FINLAND Reviewers: Professor Mauri Aalto, Ph.D., M.D. School of Medicine University of Tampere TAMPERE FINLAND Adjunct Professor Anni-Maija Linden, Ph.D. Department of Pharmacology, Faculty of Medicine University of Helsinki HELSINKI FINLAND Opponent: Chief Lorenzo Leggio, Ph.D., M.D., M.Sc. Section on Clinical Psychoneuroendocrinology and Neuropsychopharmacology NIAAA and NIDA National Institutes of Health BETHESDA, MD UNITED STATES OF AMERICA IV V Kärkkäinen, Olli Post-Mortem Brains of Alcoholics: Changes in the Glutamatergic, Serotonergic, Endocannabinoid and Neuroactive Steroid Systems University of Eastern Finland, Faculty of Health Sciences Publications of the University of Eastern Finland. Dissertations in Health Sciences 333. 2016. 99 p. ISBN (print): 978-952-61-2051-5 ISBN (pdf): 978-952-61-2052-2 ISSN (print): 1798-5706 ISSN (pdf): 1798-5714 ISSN-L: 1798-5706 ABSTRACT Alcohol (ethanol) consumption is one of the leading risk factors for many serious diseases. The pharmacology of ethanol is extremely complex, affecting many of the signaling systems not only in the brain but widely throughout the body. Alcoholics are a heterogeneous group of subjects suffering a wide spectrum of problems. Cloninger’s typology divides the spectrum of alcoholics into two subgroups: anxiety-prone late onset type 1 alcoholics and early onset, impulsive and antisocial type 2 alcoholics. Here the post-mortem brain samples of Cloninger type 1 (N=9) and type 2 (N=8) alcoholics have been studied and compared to non-alcoholic controls (N=10). The first sub-study evaluated [3H]AMPA binding to AMPA receptors. Increased binding was observed in the anterior cingulate cortex of type 2 alcoholics in comparison with controls. This elevated [3H]AMPA binding could be associated with increased impulsivity in these individuals. The second study investigated the endocannabinoid levels in the post-mortem samples of hippocampus and amygdala. Increased docosahexaenoylethanolamide levels were observed in late-onset type 1 alcoholics in the amygdala. Furthermore, a negative correlation was observed between anandamide levels and previously published metabotropic glutamate receptor 1/5 levels in the hippocampus in type 1 alcoholics, but not in controls or type 2 alcoholics. These observations could be associated with the transient receptor potential vanilloid type 1 mediated synaptic plasticity which is dependent on metabotropic glutamate receptor 5 and anandamide function. In the third study, [3H]citalopram binding to serotonin transporters was measured in brain regions associated with social cognition. Decreased [3H]citalopram binding was observed in the posterior cingulate cortex and posterior insula in all alcoholics when compared to non- alcoholic controls. Furthermore, decreased [3H]citalopram binding in the parahippocampal gyrus was seen only in the antisocial type 2 alcoholics. The decreased serotonin transporter binding in alcoholics could be associated with altered social cognitive processes. The fourth study examined levels of neuroactive steroids in the post-mortem brain samples. Increased dehydroepiandrosterone levels were seen in all alcoholics compared to controls. There were also negative correlations detected between pregnenolone levels and the previously published [3H]naloxone binding to μ-opioid receptors and similarly, increased pregnenolone levels were observed only in a sub-group of alcoholics with decreased [3H]naloxone binding in comparison with the controls. Overall, the findings of the present thesis improve our understanding of the differences between the brains of alcoholics and controls. Furthermore, they highlight the need to recognize the spectrum of alcoholics in research which hopefully will be translated into improvements in the treatment of alcoholism. National Library of Medicine Classification: QV 84, QZ 59, WL 104, WL 300, WL 348, WM 274 Medical Subject Headings: Alcoholism/pathology; Alcoholics; Brain/pathology; Amygdala; Hippocampus; Receptors, AMPA; Endocannabinoids; Serotonin; Cognition; Steroids; Dehydroepiandrosterone; Pregnenolone VI VII Kärkkäinen, Olli Alkoholistien post-mortem aivot: muutokset glutamatergisessä, serotonergisessä, endokannabinoidien, ja neuroaktiivisten steroidien järjestelmissä Itä-Suomen yliopisto, terveystieteiden tiedekunta Publications of the University of Eastern Finland. Dissertations in Health Sciences 333. 2016. 99 s. ISBN (nid.): 978-952-61-2051-5 ISBN (pdf): 978-952-61-2052-2 ISSN (nid.): 1798-5706 ISSN (pdf): 1798-5714 ISSN-L: 1798-5706 TIIVISTELMÄ Alkoholin (etanoli) käyttö on riskitekijä moniin vakaviin sairauksiin. Alkoholin farmakologia on monimutkaista ja etanoli vaikuttaakin moniin aivojen viestinvälitysjärjestelmiin. Alkoholistit ovat heterogeeninen joukko, joilla on paljon ongelmia. Cloningerin typologiassa alkoholistit jaetaan kahteen alaluokkaan. Cloningerin tyypin 1 alkoholisteilla alkoholismi alkaa myöhäisellä iällä ja he ovat taipuvaisia ahdistuneisuuteen Epäsosiaalisilla ja impulsiivisilla tyypin 2 alkoholisteilla alkoholismi puhkeaa jo nuorella iällä. Tässä väitöskirjassa tutkittiin tyypin 1 (N=9) ja tyypin 2 (N=8) alkoholistien post-mortem aivokudosnäytteitä verrattuna kontrollien näytteisiin (N=10). Ensimmäisessä osatutkimuksessa mitattiin [3H]AMPA:n sitoutumista AMPA- reseptoreihin. Tyypin 2 alkoholisteilla havaittiin korkeampi [3H]AMPA sitoutuminen anteriorisessa pihtipoimussa kontrolleihin verrattuna. Tämä muutos saattaa liittyä heidän impulsiiviseen luonteeseensa. Toisessa osatutkimuksessa mitattiin endokannabinoiditasoja amygdala ja hippokampus alueiden aivonäytteistä. Tyypin 1 alkoholisteilla havaittiin kohonneet docosahexaoneylamide-tasot verrattuna kontrolleihin. Tyypin 1 alkoholistien hippokampuksessa havaittiin myös negatiivinen korrelaatio anandamidikonsentraatioiden ja metabotropisten glutamaattireseptorien 1/5 – tasojen välillä. Nämä tulokset voivat