MANEJO INTEGRADO DE ENFERMEDADES Y PLAGAS
SELECTIVIDAD DEL COMMAND EN PREEMERGENCIA EN EL PASO 144 Néstor Saldain 1/ , Enrique Deambrosi 1/
The overall goal of this master’s thesis research was to gain a better understanding of how adolescent binge drinking affects anxiety-related peptide, CRF using a voluntary alcohol self-administration model. Another goal of this thesis was to determine possible prescreening methods for alcohol intake during adolescence. Experiments herein demonstrated that the behavioral traits and molecular processes of anxiety are directly linked to alcohol intake during adolescence. We provided evidence that chronic binge drinking during adolescence has long-lasting effects on key regulators of the stress system, specifically the CeA. In particular, CRF-ir in the CeA is down-regulated as a result of binge drinking during adolescence and lasts well into adulthood. Interestingly, vapor-induced dependence in adulthood does not exacerbate the binge effect in the CeA. Social anxiety-like behaviors were measured before animals were exposed to alcohol to try and account for the individual variations of intake in the voluntary self-administration model. Using the social interaction test, frequency of self-grooming was found to be negatively correlated with alcohol intake. In other words, the more animals displayed this non-social behavior, the less alcohol they consumed during adolescence.
Everyday stressors can greatly affect the functioning of CRF-ir cells and the release of the peptide, leading to acute and long-term changes of the HPA system and changes in behavioral traits. Data in this thesis indicate that alcohol affects CRF peptide activity in the CeA. However, it is important to note that
55
CRF measurements throughout this thesis were for CRF peptide via immunolabeling. Histochemical labeling was nickel-enhanced to ensure that as we labeled as much CRF peptide as possible. Labeling involved both cell bodies and processes, however analysis only included cell bodies. Therefore, the decrease in CRF-ir cell number does not take into account total immunoreactivity (i.e. CRF-ir cell bodies and processes). Also, as previously mentioned, expression of peptide is not indicative of CRF mRNA levels and in situ hybridization is necessary to determine the effects of binge drinking on mRNA (see Chapter 3, page 31).
CRF cells control the entire stress system through their regulation of the HPA axis, however little is known about effects of alcohol on this system in females. Experiment 2 of this thesis also introduced the effects of alcohol on the
female stress system. Females are twice as likely to develop stress-related
psychiatric disorders (Bangasser and Valentino, 2012). It is known that gonadal hormones play a role in this sex difference in stress-related disorders. In fact, gonadal hormones can regulate CRF gene expression via estrogen and androgen response elements located at the promoter region of the CRF gene (Bao et al., 2006; Vamvakopoulos and Chrousos, 1993). Estrogen stimulates CRF expression, while androgens suppress CRF expression (Bohler et al., 1990; Figueiredo et al., 2007; Lund et al., 2004). Future studies can examine the effects of peripubertal gonadectomy on the CRF system’s response to adolescent binge drinking. This type of study will help to better understand the
56
link between pubertal development, alcohol intake during adolescence and the response of the stress system.
Self-grooming in rodents is depicted as a response to anxiogenic and stressful stimuli. Neurochemically, it is known that dopamine release is significantly lower in the substantia nigra, medial prefrontal cortex, and amygdala in animals that display high self-grooming behavior when presented with a stressful situation (Homberg et al., 2002). However, Homberg et al. (2002) found opposite effects compared to the findings in this thesis. Specifically, higher frequency of self-grooming was positively correlated with the self-administration of cocaine, whereas in the present study it was negatively correlated with the self-administration of alcohol. Still, it is important to note, that alcohol may not activate the same pathway in the reward system.
The ability to prescreen for alcohol intake for the will allow for better control in alcohol exposure and will account for the individual variations of intake while maintaining the integrity of the voluntary aspect of the operant self- administration model. Prescreening for intake can also allow for further investigation in certain measurements that are otherwise unattainable. For example, measuring CRF-ir in the CeA is not possible to do before alcohol exposure. By prescreening animals, animals that are potentially heavier drinkers can be capped at different levels of access to alcohol and better controlling the overall intake of each animal. Capping at different levels can be used to determine if the effect of alcohol on CRF-ir in the CeA is dose-dependent.
57
One limitation includes the inability to further examine what is happening to the CRF-ir cells on a morphological or molecular level. Is the number of CRF- ir cells found in the adolescent and adult CeA a true representation of a decrease in cell number or have some cells not been labeled because the peptide was already released? If it was a true decrease in cell number and not a decrease in peptide within the cells, was it due to alcohol-induced decrease in neurogenesis or increase in cell death? The research conducted for this thesis has opened the doors to further exploring the effects of alcohol on the CRF system. Future directions could include measuring CRF mRNA levels via in situ hybridization to get a true representation of the peptide gene expression. Also, examining markers for apoptosis, such as Caspase-3, to get a better idea of what is happening to these cells after exposure to alcohol. Interestingly, CRF mRNA and immunreactivity is down-regulated in rats that were selectively bred to prefer alcohol (Hwang et al., 2004). This effect occurs before animals are even exposed to alcohol. A future study could include repeating the experiments using P rats to determine if adolescent binge drinking and adult dependence further exacerbates this down-reguation in CRF. It would also be important to investigate the effects of adolescent drinking and adult dependence on the CeA CRF system in females, as the long-term consequences of alcohol may differ with sex.
In summary, the work presented in this thesis conveys an overall theme about the link between alcohol and anxiety-related molecular and behavioral responses. CRF is a small part of a much larger and complicated system and is
58
easily influenced by both endogenous and exogenous stimuli. As one of the most used and abused drugs among adolescents today, alcohol is one of the most prevalent stressor that indirectly and directly affects the stress system. Further investigation of the effects of alcohol on the function and regulation of the stress system the behavioral traits that could be linked to risk-seeking behavior could enhance pharmacological therapies and preventative measures for addiction.
59
REFERENCES
Adinoff B, Junghanns K, Kiefer F, Krishnan-Sarin S (2005) Suppression of the HPA axis stress-response: implications for relapse. Alcohol Clin Exp Res 29:1351-1355.
Almeida OFX, Soaib M, Deicke J, Fischer D, Darwish MH, Patchev VK (1998) Gender differences in ethanol preference and ingestion in rats. J Clin Invest 101:2677-2685.
American Academy of Pediatrics Binge Drinking, Washington, D.C, 1999.
Arnett J (1992) Reckless behavior in adolescence: a developmental perspective. Developmental Review 12:339-373.
Bale TL, Vale WW (2004)CRF and CRF receptors: role in stress responsivity and other behaviors. Annu. Rev. Pharmacol. Toxicol. 44:525–57.
Ballard HS (1997) The hematological complication of alcoholism. Alc Health Res
World 21:42-52.
Bibb JL, Chambless DL (1986) Alcohol use and abuse among diagnosed agoraphobics. Behav Res Ther 24:49–58.
Binder EB, Nemeroff CB (2009) The CRF system, stress, depression and anxiety – insights from human genetic studies. Molecular Psychiatry 1-15.
Bradley KA, Badrinath S, Bush K, Boyd-Wickizer J, Anawalt B (1998) Medical risks for women who drink alcohol. J Gen Intern Med 13:675-639.
Breese GR, Overstreet DH, Knapp DJ (2005) Conceptual framework for the etiology of alcoholism: a “kindling”/stress hypothesis. Psychopharm 178:367- 380.
Breu J, Touma C, Holter SM, Knapman A, Wurst W, Deussing JM (2012) Urocortin 2 modulates aspects of social behaviour in mice. Behavioural Brain
Research 233:331-336.
Brook JS, Whiteman M, Finch SJ (1992) Childhood aggression, adolescent delinquency and drug use: a longitudinal study. Journal of Genetic
Psychology 153:369–383.
Brook JS, Whiteman M, Finch SJ, Cohen P (1996) Young adult drug use and delinquency: childhood antecedents and adolescent mediators. J Amer Acad
60
Brooks-Gunn J, Attie I (1996) Devlepmental psychopathology in the context of adolescence. In: Lenzenweger MF, Haugaard JJ, editors. Frontiers of developmental psychopathology, New York, NY: Oxford University Press. p. 148-189.
Caldwell E, Riccio D (2010) Alcohol self-administration in rats: modulation by temporal parameters related to related mild social defeat stress. Alcohol 44:265-274.
Cappell H, Herman CP (1972) Alcohol and tension reduction. A review. Q J Stud
Alcohol 33:33–64.
Choi S, Weisberg SN, Kellogg CK (1997) Control of endogenous norepinephrine release in the hypothalamus of male rats changes over adolescent development. Brain Res Dev Brain Res 98:134-141.
Clark PA (1998) Puberty: When it comes too soon. Guidelines for the Evaluation of Sexual Precocity. KMA J 96:441-447.
Colbern D (1981) Water immersion, excessive grooming, and paper shredding in the rat. Behav Neural Biol 32:428-437.
Compas BE, Orosan PG, Grant KE (1993) Adolescent stress and coping: implications for psychopathology durng adolescence. J Adol 16:331-349. Cools AR, Wiegant VM, Gispen WH (1978) Distinct dopaminergic systems in
ACTH induced grooming. Eur J Pharm 50:265-268.
Cornelius JR, Bukstein O, Salloum I, Clark D (2003) Alcohol and psychiatric comorbidity. Recent Dev Alcohol 16:361–374.
Courtney KE, Polich J (2009) Binge drinking in young adults: Data, definitions, and determinants. Psychol Bull 135:142-156.
Cox KH, Rissman EF (2011) Sex differences in juvenile mouse social behavior are influenced by sex chromosomes and social context. Genes Brain Behav 10: 465-472.
Crews FT, Braun CJ, Hoplight B, Switzer RC 3rd, Knapp DJ (2000) Binge ethanol
consumption causes differential brain damage in young adolescent rats compared with adult rats. Alcohol Clin Exp Res 24:1712-1723.
Crews FT, Buckley T, Dodd PR, Ende G, Foley N, Harper C, He J, Innes D, Loh EW, Pfefferbaum A, Zou J, Sullivan EV (2005) Alcohol neurobiology: Changes in dependence and recovery. Alcohol Clin Exp Res 29:1504-1513.
61
Crum RM, Pratt LA (2001) Risk of heavy drinking and alcohol use disorders in social phobia: A prospective analysis. American Journal of Psychiatry 158: 1693-1700.
Csikszentmihalyi M, Larson R, Prescott S (1977) The ecology of adolescent activity and experience. Journal of Youth and Adolescence 6:281-294.
Dautzenberg FM, Hauger RL (2002) The CRF peptide family and their receptors: yet more partners discovered. Trends Pharmacol Sci 23:71-77.
Deroche-Gamonet V, Sillaber I, Aouizerate B, Izawa R, Jaber M, Ghozland S, Kellendonk C, Le Moal M, Spanagel R, Schutz G, Tronche F, Piazza PV (2003) The glucocorticoid receptor as a potential target to reduce cocaine abuse. J Neurosci 23:4785-4790.
de Wit H, Soderpalm AH, Nikolayev L, Young E (2003) Effects of acute social stress on alcohol consumption in healthy subjects. Alcohol Clin Exp Res 27:1270-1277.
Doremus TL, Brunell SC, Varlinskaya EI, Spear LP (2003) Anxiogenic effects during withdrawal from acute ethanol in adolescent and adult rats.
Pharmacol Biochem Behav 75:411-418.
Doremus TL, Brunell SC, Rajendran P, Spear LP (2005) Factors influencing elevated ethanol consumption in adolescent relative to adult rats. Alcohol
Clin Exp Res 29:1796-1808.
Drago F, Contarino A, Busa L (1999) The expression of neuropeptide-induced
excessive grooming behavior in dopamine D1 and D2 receptor-deficient mice.
Eur J Pharmacol 365: 125-131.
Draski LJ, Bice PJ, Deitrich RA (2001) Developmental alteration of ethanol sensitivity in bred high and low alcohol sensitive rats. Pharmacol Biochem
Behav 70:387-396.
Duka T, Townshend JM, Collier K, Stephens DN (2003) Impairment in cognitive functions after multiple detoxifications in alcohol inpatients. Alcohol Clin Exp
Res 27:1563-1572.
Dunn AJ, Berridge CW, Lai YI, Yachabach TL (1986) CRF-induced excessive grooming behavior in rats and mice. Peptides 8:841-844.
Dunn AJ, Berridge CW (1990) Physiological and behavioral responses to corticotropin-releasing factor administration: is CRF a mediator of anxiety or stress response? Brain Res Rev 15:71-100.
62
Equibar JR, Romero-Carbente JC, Moyaho A (2003) Behavioral differences
between selectively bred rats: D1 versus D2 receptors in yawning and
grooming. Pharm, Biochem and Behavior 74:827-832.
Erickson CK (1984) Ethanol clearance in nine inbred rat strains. Alc Clin Exp Res 8:491-494.
Erikkson K (1969) Effects of ovariectomy and contraceptive hormones on voluntary alcohol consumption in the albino rat. J Stud Alcohol 4:1-5
Faravelli C, Lo Sauro C, Lelli L, Pietrini F, Lazzeretti L, Godini L, Benni L, Fioravanti G, Talamba GA, Castellini G, Ricca V (2012) The role of life events and HPA axis in anxiety disorders: a Review. Curr Pharm Des, in press.
Grigoriadis DE, De Souza EB (1989) Heterogeneity between brain and pituitary corticotropin-releasing factor receptors is due to differential glycosylation.
Endocrinology 125:1877-1887.
Farber NB, Wozniak DF, Price MT, Labruyere J, Hus J, St. Peter H, Plney JW (1995) Age-specific neurotoxicity in the rat associated with NMDA receptor blockade: potential relevance to schizphrenia? Biological Psychiatry 38:788- 796.
File SE (1980) The use of social interaction as a method for detecting anxiolytic activity of chlordiazepoxide-like drugs. Journal of Neuroscience Methods 2: 219-238.
File SE, Seth P (2003) A review of 25 years of the social interaction test.
European Journal of Pharmacology 463:35-53.
Finn DA, Belknap JK, Cronise K, Yoneyama N, Murillo A, et al. (2005) A procedure to produce high alcohol intake in mice. Psychopharmacology
(Berl) 178: 471–480.
Forger NG, Morin LP (1982) Reproductive state modulates ethanol intake in rats: Effects of ovariectomy, ethanol concentration, estrous cycle and pregnancy.
Pharmacol Biochem Behav 17:323-331.
Freeza M, Di Padova C, Pozzato G, Terpin M, Baraona E, Lieber CS (1990) High blood alcohol levels in women. The role of decreased gastric alcohol dehydrogenase activity and first-pass metabolism. N Engl J Med 322:95-99.
63
Frias J, Rodriguez R, Torres JM, Ruiz E, Ortega E (2000) Effects of acute alcohol intoxication on pituitary-gonadal axis hormones, pituitary-adrenal axis hormones, b-endorphin and prolactin in human adolescents of both sexes.
Life Sciences 67:1081-1086.
Frias J, Torres JM, Miranda MT, Ruiz E, Ortega E (2002) Effects of acute alcohol intoxication on pituitary-gonadal axis hormones, pituitary-adrenal axis hormones, b-endorphin and prolactin in human adults of both sexes. Alcohol
Alcohol 37:169-173.
Funk CK, O’Dell LE, Crawford EF, Koob GF (2006) Corticotropin-releasing factor within the central nucleus of the amygdala mediates enhanced ethanol self- administration in withdrawal, ethanol-dependent rats. J Neurosci 26:11324- 11332.
Funk CK, Zorrilla EP, Lee MJ, Rice KC, Koob GF (2007) Corticotropin-releasing factor 1 antagonists selectively reduce ethanol self-administration in ethanol- dependent rats. Biological Psychiatry 61:78-86.
Gaillard RC (2003) Interactions between the immune and neuroendocrine systems: clinical implications. J Soc Biol 197:89-95.
Gatch MB, Wallis CJ, Lal H (1999) Effects of NMDA antagonists on ethanol- withdrawal induced "anxiety" in the elevated plus maze. Alcohol 19:207–211. Geracioti TD Jr, Loosen PT, Ebert MH, Ekhator NN, Burns D, Nicolson WE, Orth
DN (1994) Concentrations of corticotropin-releasing hormone, norepinephrin, MHPG, 5-hydroxyindoleacetic acid, and tryptophan in the cerebrospinal fluid of alcoholic patients: Serial sampling studies. Neuroendocrinology 60:635- 642.
Gilpin NW, Richardson HN, Cole M, Koob GF (2008) Vapor inhalation of alcohol in rats. Curr Protoc Neurosci Chapter 9: Unit 9 29.
Gilpin NW, Smith AD, Cole M, Weiss F, Koob GF (2009) Operant behavior and alcohol levels in blood and brain of alcohol-dependent rats. Alcohol Clin Exp
Res 33:2113-2123.
Gilpin NW (2012) Corticotropin-releasing factor (CRF) and neuropeptide Y (NPY): effects on inhibitory transmission in central amygdala, and anxiety- & alcohol-related behaviors. Alcohol 46:329-337.
Gilpin NW, Karanikas CA, Richardson HN (2012) Adolescent binge drinking leads to changes in alcohol drinking, anxiety, and amygdalar corticotropin releasing factor cells in the adulthood in male rats. PlosOne 7(2), e31466.
64
Gispen WH, Isaacson RL (1981) ACTH-induced excessive grooming in the rat.
Pharmacol Ther 12:209-246.
Goeders NE (2002) The HPA axis and cocaine reinforcement.
Psychoneuroendocrinology 27:13-33.
Grant EC, Mackintosh JH (1963) A comparison of the social postures of some common laboratory rodents. Behaviour 21:246-259.
Grant BF, Dawson DA, Stinson FS, Chou SP, Dufour MC, Pickering RP (2004) The 12-month prevalence and trends in DSM-IV alcohol abuse and dependence: United States, 1991-1992 and 2001-2002. Drug Alcohol
Depend 74:223-234.
Hartford TC, Grant BF, Yi H-Y, Chen CM (2005) Patterns of DSM-IV alcohol abuse and dependence criteria among adolescents and adults: results from the 2001 National Household Survey on Drug Abuse. Alcohol Clin Exp Res 29:810-828.
Hasin D, Van Rossem R, McCloud S, Endicott J (1997) Alcohol dependence and abuse diagnoses: validity in community sample heavy drinkers, Alcohol Clin
Exp Res 21:213–219.
Heilig M, Koob GF (2007) A key role for corticotropin-releasing factor in alcohol dependence. Trends in Neuroscience 30: 399-406.
Heim C, Nemeroff CB (2001) The role of childhood trauma in the neurobiology of mood and anxiety disorders: preclinical and clinical studies. Biol Psychiatry 49:1023-1039.
Herman JP, Ostrander MM, Mueller NK, Figueiredo H (2005) Limbic system mechanisms of stress regulation: Hypothalamo-pituitary-adrenocortical axis.
Prog Neuropsychopharmacol Biol Psychiatry 29:1201-13.
Hill KG, White HR, Chung IJ, Hawkins JD, Catalano RF (2000) Early adult outcomes of adolescent binge drinking: person- and variable-centered analyses of binge drinking trajectories. Alcohol Clin Exp Res 24: 892–901. Holter SM, Engelmann M, Kirschke C, Liebsch G, Landgraf R, Spanagel R
(1998) Long-term ethanol self-administration with repeated ethanol deprivation episodes changes ethanol drinking pattern and increases anxiety-related behaviour during ethanol deprivation in rats. Behav
65
Homberg JR, van den Akker M, Raaso HS, Wardeh G, Binnekade R, Schoffelmeer ANM, de Vries TJ (2002) Enhanced motivation to self- administer cocaine is predicted by self-grooming behaviour and relates to dopamine release in the rat medial prefrontal cortex and amygdala. Eur J
Neurosci 15: 1542-1550.
Huang MM, Overstreet DH, Knapp DJ, Angel R, Wills TA, Navarro M, Rivier J, Vale W, Breese GR (2010) Corticotropinn-releasing factor (CRF) sensitization of ethanol withdrawal-induced anxiety-like behavior is brain site specific and mediated by CRF-ir receptors: relation to stress-induced sensitization. J Pharmacol Exp Ther 332:298-307.
Hwang BH, Stewart R, Zhang JK, Lumeng L, Li TK (2004) Corticotropin-releasing factor gene expression is down-regulated in the central nucleus of the amygdala of alcohol-preferring rats which exhibit high anxiety: a comparison between rat lines selectively bred for high and low alcohol preference. Brain
Res 1026:143-150.
Irvine EE, Bagnalasta M, Marcon C, Motta C, Tessari M, File SE, Chiamulera C (2001) Nicotine self-administration and withdrawal: modulatory of anxiety I the social interaction test in rats. Psychopharmacology (Berl). 153:315-320. Jang KL, Livesley WJ, Vernon PA (1997) Gender-specific etiology differences in
alcohol and drug problems: a behavioural genetic analysis. Addiction 92:1265-1276.
Johnston AL, File SE (1991) Sex differences in animal tests of anxiety. Physiol
Behav 49:245-250.
Johnston LD, O’Malley PM, Bachman JG, Schulenberg JE (2006) Monitoring the future national results on adolescent drug use: overview of key findings, 2005. Ntl Inst on Drug Abuse.
Johnston LD, O’Malley PM, Backman JG, Schulenberg JE (2007) Monitoring the future national results on adolescent drug use: overview of key findings, 2006. In. Bethesda, MD.
Jolles J, Rompa-Barendregt J, Gispen WH (1979) ACTH-Induced excessive grooming in the rat; influence of environmental and motivational factors.
Horm Behav 12:60-72.
Kametani H (1988) Analysis of age related changes in stress-induced grooming in the rat. Ann NY Acad Sci 525:101-113.
Keller-Wood ME, Dallman MF (1984) Corticosteroid inhibition of ACTH secretion.
66
Kiess W, Meidert A, Dressendorfer RA, Schriever K, Kessler U, Konig A, Schwartz HP, Strasburger CJ (1995) Salivary cortisol-levels throughout childhood and adolescence – relation with age, pubertal stage, and weight.
Pediatr Res 37: 502-506.
Koob, G. (2003) Alcoholism: allostasis and beyond. Alcohol Clin Exp Res 27:232- 243.
Koob, G. (2008) A role for brain stress systems in addiction. Neuron 59:11-34. Korenbrot CC, Huhtaniemi IT, Weiner RI (1977) Preputial separation as an
external sign of pubertal development in the male rat. Biol Reproduction 17:298-303.
Lancaster FE (1994) Gender differences in the brain: implications for the study of human alcoholism. Alc Clin Exp Res 18:740-746.
Lancaster FE, Brown TD, Coker KL, Elliot JA, Wren SB (1996) Sex differences in alcohol preference and drinking patterns emerge during the early postpubertal period in Sprague-Dawley rats. Alc Clin Exp Res 20:1043-1049. Laviola G, Adriani W, Terranova ML, Gerra G (1999) Psychobiological risk
factors for vulnerability to psychostimulants in human adolescents and animal models. Neurosci Biobehav Rev 23:993-1010.
Laviola G, Adriani W, Morley-Fletcher S, Terranova M (2002) Peculiar response of adolescent mice to acute and chronic stress and to amphetamine: evidence of sex differences. Behavioral Brain Research 130:117-125.
Liu X, Mao X, Change SL (2011) Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking. Int J Clin
Exp Med 4:252-257.
Larkin JW, Binks SL, Li Y, Selvage D (2010) The Role of oestradiol in sexually dimorphic hypothalamic-pituitary-adrenal axis responses to Intracerebroventricular ethanol administration in the rat. J