Skip to main content

Advertisement

SpringerLink
Log in

Effects of different ethanol-administration regimes on mRNA and protein levels of steroid 5α-reductase isozymes in prefrontal cortex of adolescent male rats

  • Original Investigation
  • Published:
Psychopharmacology Aims and scope Submit manuscript

Abstract

Rationale

Underage drinking is a leading public health problem in developed countries. An increasing proportion of adolescents consume alcoholic beverages every weekend. Increased anxiety, irritability, and depression among adolescents may induce them to seek for the anxiolytic and rewarding properties of alcohol. Allopregnanolone (AlloP) shares rewarding effects of ethanol and modulates ethanol intake. The rate-limiting enzyme in the biosynthesis of AlloP is steroid 5α-reductase (5α-R), which is expressed as three isozymes, 5α-R1, 5α-R2, and 5α-R3.

Objective

The objective of this study was to quantify the expression levels of 5α-R isozymes in prefrontal cortex (PFC) of adolescent male rats after three different regimes of ethanol administration.

Methods

Adolescent male Wistar rats were administered with ethanol (4 g/kg) or saline intraperitoneally for 1 day (acute), for 7 days (chronic), or every 72 h for 14 days (chronic intermittent). Messenger (m)RNA and protein levels of 5α-R isozymes were measured by quantitative RT-PCR and Western blot, respectively.

Results

Ethanol significantly increased mRNA and protein levels of 5α-R1, 5α-R2, and 5α-R3 in the three different regimes of ethanol administration, being higher in the chronic intermittent regime in comparison with the others.

Conclusions

The expression of the AlloP-biosynthetic enzyme 5α-Rs increases in the prefrontal cortex of adolescent male rats under acute, chronic, and chronic intermittent regime of ethanol administration. The latter is very interesting because it mimics the teenage drinking behavior.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Bailey SL, Rachal JV (1993) Dimensions of adolescent problem drinking. J Stud Alcohol 54:555–565

    CAS  PubMed  Google Scholar 

  • Barbaccia ML, Affricano D, Trabucchi M, Purdy RH, Colombo G, Agabio R, Gessa GL (1999) Ethanol markedly increases “GABAergic” neurosteroids in alcohol-preferring rats. Eur J Pharmacol 384:R1–R2

    Article  CAS  PubMed  Google Scholar 

  • Bonomo YA, Bowes G, Coffey C, Carlin JB, Patton GC (2004) Teenage drinking and the onset of alcohol dependence: a cohort study over seven years. Addiction 99:1520–1528

    Article  PubMed  Google Scholar 

  • Bradford MN (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Boyd KN, Kumar S, O’Buckley TK, Morrow AL (2010) Chronic ethanol exposure produces tolerance to elevations in neuroactive steroids: mechanisms and reversal by exogenous ACTH. J Neurochem 115:142–152

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Broadwater M, Varlinskaya EI, Spear LP (2011) Chronic intermittent ethanol exposure in early adolescent and adult male rats: effects on tolerance, social behavior, and ethanol intake. Alcohol Clin Exp Res 35:1392–1403

    CAS  PubMed Central  PubMed  Google Scholar 

  • Cagetti E, Pinna G, Guidotti A, Baicy K, Olsen RW (2004) Chronic intermittent ethanol (CIE) administration in rats decreases levels of neurosteroids in hippocampus, accompanied by altered behavioral responses to neurosteroids and memory function. Neuropharmacology 46:570–579

    Article  CAS  PubMed  Google Scholar 

  • Cantagrel V, Lefeber DJ, Ng BG et al (2010) SRD5A3 is required for converting polyprenol to dolichol and is mutated in a congenital glycosylation disorder. Cell 142:203–217

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Castro B, Sánchez P, Torres JM, Ortega E (2013) Effects of adult exposure to bisphenol A on genes involved in the physiopathology of rat prefrontal cortex. PLoS One 8:e73584

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Cook JB, Dumitru AM, O’Buckley TK, Morrow AL (2014) Ethanol administration produces divergent changes in GABAergic neuroactive steroid immunohistochemistry in the rat brain. Alcohol Clin Exp Res 38:90–99

    Article  CAS  PubMed  Google Scholar 

  • Devaud LL, Fritschy JM, Morrow AL (1998) Influence of gender on chronic ethanol-induced alterations in GABAA receptors in rats. Brain Res 15:222–230

    Google Scholar 

  • Devaud LL, Purdy RH, Finn DA, Morrow AL (1996) Sensitization of gamma-aminobutyric acidA receptors to neuroactive steroids in rats during ethanol withdrawal. J Pharmacol Exp Ther 278:510–517

    CAS  PubMed  Google Scholar 

  • Eva C, Mele P, Collura D, Nai A, Pisu MG, Serra M, Biggio G (2008) Modulation of neuropeptide Y and Y1 receptor expression in the amygdala by fluctuations in the brain content of neuroactive steroids during ethanol drinking discontinuation in Y1R/LacZ transgenic mice. J Neurochem 104:1043–1054

    Article  CAS  PubMed  Google Scholar 

  • Finn DA, Sinnott RS, Ford MM, Long SL, Tanchuck MA, Phillips TJ (2004) Sex differences in the effect of ethanol injection and consumption on brain allopregnanolone levels in C57BL/6 mice. Neuroscience 123:813–819

    Article  CAS  PubMed  Google Scholar 

  • Ford MM, Nickel JD, Phillips TJ, Finn DA (2005a) Neurosteroid modulators of GABAA receptors differentially modulate ethanol intake patterns in male C57BL/6 J mice. Alcohol Clin Exp Res 29:1630–1640

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ford MM, Nickel JD, Finn DA (2005b) Treatment with and withdrawal from finasteride alter ethanol intake patterns in male C57BL/6 J mice: potential role of endogenous neurosteroids? Alcohol 37:23–33

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ford MM, Yoneyama N, Strong MN, Fretwell A, Tanchuck M, Finn DA (2008) Inhibition of 5alpha-reduced steroid biosynthesis impedes acquisition of ethanol drinking in male C57BL/6 J mice. Alcohol Clin Exp Res 32:1408–1416

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Fröjd S, Ranta K, Kaltiala-Heino R, Marttunen M (2011) Associations of social phobia and general anxiety with alcohol and drug use in a community sample of adolescents. Alcohol Alcohol 46:192–199

    Article  PubMed  Google Scholar 

  • Fronhoffs S, Totzke G, Stier S, Wernert N, Rothe M et al (2002) A method for the rapid construction of cRNA standard curves in quantitative real-time reverse transcription polymerase chain reaction. Mol Cell Probes 16:99–110

    Article  CAS  PubMed  Google Scholar 

  • Gessa GL, Serra S, Vacca G, Carai MA, Colombo G (2005) Suppressing effect of the cannabinoid CB1 receptor antagonist, SR147778, on alcohol intake and motivational properties of alcohol in alcohol-preferring sP rats. Alcohol Alcohol 40:46–53

    Article  CAS  PubMed  Google Scholar 

  • Golka K, Wiese A (2004) Carbohydrate-deficient transferrin (CDT)–a biomarker for long-term alcohol consumption. J Toxicol Environ Health B Crit Rev 7:319–337

    Article  CAS  PubMed  Google Scholar 

  • Grobin AC, Matthews DB, Devaud LL, Morrow AL (1998) The role of GABA(A) receptors in the acute and chronic effects of ethanol. Psychopharmacology 139:2–19

    Article  CAS  PubMed  Google Scholar 

  • Heilig M, Egli M, Crabbe JC, Becker HC (2010) Acute withdrawal, protracted abstinence and negative affect in alcoholism: are they linked? Addict Biol 15:169–184

    Article  PubMed Central  PubMed  Google Scholar 

  • Irwig MS (2013) Decreased alcohol consumption among former male users of finasteride with persistent sexual side effects: a preliminary report. Alcohol Clin Exp Res 37:1823–1826

    Article  CAS  PubMed  Google Scholar 

  • Janis GC, Devaud LL, Mitsuyama H, Morrow AL (1998) Effects of chronic ethanol consumption and withdrawal on the neuroactive steroid 3alpha-hydroxy-5alpha-pregnan-20-one in male and female rats. Alcohol Clin Exp Res 22:2055–2061

    CAS  PubMed  Google Scholar 

  • Kim HJ, Ha M, Park CH, Park SJ, Youn SM, Kang SS, Cho GJ, Choi WS (2003) StAR and steroidogenic enzyme transcriptional regulation in the rat brain: effects of acute alcohol administration. Brain Res Mol Brain Res 115:39–49

    CAS  PubMed  Google Scholar 

  • Koob GF (2003) Alcoholism: allostasis and beyond. Alcohol Clin Exp Res 27:232–243

    Article  CAS  PubMed  Google Scholar 

  • Kumar S, Porcu P, Werner DF, Matthews DB, Diaz-Granados JL, Helfand RS, Morrow AL (2009) The role of GABA(A) receptors in the acute and chronic effects of ethanol: a decade of progress. Psychopharmacology (Berl) 205:529–564

    Article  CAS  Google Scholar 

  • Kuntsche E, Knibbe R, Gmel G, Engels R (2005) Why do young people drink? A review of drinking motives. Clin Psychol Rev 2:841–861

    Article  Google Scholar 

  • Lenz B, Schöpp E, Müller CP, Bleich S, Hillemacher T, Kornhuber J (2012) Association of V89L SRD5A2 polymorphism with craving and serum leptin levels in male alcohol addicts. Psychopharmacology 224:421–429

    Article  CAS  PubMed  Google Scholar 

  • Makridakis N, Akalu A, Reichardt JK (2004) Identification and characterization of somatic steroid 5alpha-reductase (SRD5A2) mutations in human prostate cancer tissue. Oncogene 23:7399–7405

    Article  CAS  PubMed  Google Scholar 

  • Matsui D, Sakari M, Sato T, Murayama A, Takada I, Kim M, Takeyama K, Kato S (2002) Transcriptional regulation of the mouse steroid 5-alpha-reductase type II gene by progesterone in brain. Nucleic Acids Res 30:1387–1393

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mellon SH, Griffin LD, Compagnone NA (2001) Biosynthesis and action of neurosteroids. Brain Res Brain Res Rev 37:3–12

    Article  CAS  PubMed  Google Scholar 

  • Morava E, Wevers RA, Cantagrel V, Hoefsloot LH, Al-Gazali L et al (2010) A novel cerebello-ocular syndrome with abnormal glycosylation due to abnormalities in dolichol metabolism. Brain 133:3210–3220

    Article  PubMed  Google Scholar 

  • Morris SA, Kelso ML, Liput DJ, Marshall SA, Nixon K (2010) Similar withdrawal severity in adolescents and adults in a rat model of alcohol dependence. Alcohol 44:89–98

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Morrow AL, Janis GC, VanDoren MJ, Matthews DB, Samson HH, Janak PH, Grant KA (1999) Neurosteroids mediate pharmacological effects of ethanol: a new mechanism of ethanol action? Alcohol Clin Exp Res 23:1933–1940

    Article  CAS  PubMed  Google Scholar 

  • Morrow AL, Porcu P, Boyd KN, Grant KA (2006) Hypothalamic-pituitary-adrenal axis modulation of GABAergic neuroactive steroids influences ethanol sensitivity and drinking behavior. Dialogues Clin Neurosci 8:463–477

    PubMed Central  PubMed  Google Scholar 

  • Morrow AL, VanDoren MJ, Penland SN, Matthews DB (2001) The role of GABAergic neuroactive steroids in ethanol action, tolerance and dependence. Brain Res Brain Res Rev 37:98–109

    Article  CAS  PubMed  Google Scholar 

  • Paul SM, Purdy RH (1992) Neuroactive steroids. FASEB J 6:2311–2322

    CAS  PubMed  Google Scholar 

  • Paxinos G, Watson C (1986) The rat brain in stereotaxic coordinates, 2nd edn. Academic, New York

    Google Scholar 

  • Pereira G, Wood L, Foster S, Haggar F (2013) Access to alcohol outlets, alcohol consumption and mental health. PLoS One 8:e53461

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Pierucci-Lagha A, Covault J, Feinn R, Nellissery M, Hernandez-Avila C, Oncken C, Morrow AL, Kranzler HR (2005) GABRA2 alleles moderate the subjective effects of alcohol, which are attenuated by finasteride. Neuropsychopharmacology 30:1193–1203

    Article  CAS  PubMed  Google Scholar 

  • Poletti A, Negri-Cesi P, Rabuffetti M, Colciago A, Celotti F, Martini L (1998) Transient expression of the 5alpha-reductase type 2 isozyme in the rat brain in late fetal and early postnatal life. Endocrinology 139:2171–2178

    CAS  PubMed  Google Scholar 

  • Quoilin C, Didone V, Tirelli E, Quertemont E (2013) Higher long-lasting ethanol sensitization after adolescent ethanol exposure in mice. Psychopharmacology Dec 7 (in press)

  • Ramaker MJ, Ford MM, Fretwell AM, Finn DA (2011) Alteration of ethanol drinking in mice via modulation of the GABA(A) receptor with ganaxolone, finasteride, and gaboxadol. Alcohol Clin Exp Res 35:1994–2007

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sánchez P, Torres JM, Olmo A, O’Valle F, Ortega E (2009) Effects of environmental stress on mRNA and protein expression levels of steroid 5alpha-Reductase isozymes in adult rat brain. Horm Behav 56:348–353

    Article  PubMed  Google Scholar 

  • Sánchez P, Torres JM, Ortega E (2005) Effects of dihydrotestosterone on brain mRNA levels of steroid 5alpha-reductase isozymes in early postnatal life of rat. Neurochem Res 30:577–581

    Article  PubMed  Google Scholar 

  • Sanna E, Talani G, Busonero F, Pisu MG, Purdy RH, Serra M, Biggio G (2004) Brain steroidogenesis mediates ethanol modulation of GABAA receptor activity in rat hippocampus. J Neurosci 24:6521–6530

    Article  CAS  PubMed  Google Scholar 

  • Shope JT, Copeland LA, Dielman TE (1994) Measurement of alcohol use and misuse in a cohort of students followed from grade 6 through grade 12. Alcohol Clin Exp Res 18:726–733

    Article  CAS  PubMed  Google Scholar 

  • Silvers JM, Tokunaga S, Mittleman G, Matthews DB (2003) Chronic intermittent injections of high-dose ethanol during adolescence produce metabolic, hypnotic, and cognitive tolerance in rats. Alcohol Clin Exp Res 27:1606–1612

    Article  CAS  PubMed  Google Scholar 

  • Silvers JM, Tokunaga S, Mittleman G, O’Buckley T, Morrow AL, Matthews DB (2006) Chronic intermittent ethanol exposure during adolescence reduces the effect of ethanol challenge on hippocampal allopregnanolone levels and Morris water maze task performance. Alcohol 39:151–158

    Article  CAS  PubMed  Google Scholar 

  • Tanchuck MA, Long SL, Ford MM, Hashimoto J, Crabbe JC, Roselli CE, Wiren KM, Finn DA (2009) Selected line difference in the effects of ethanol dependence and withdrawal on allopregnanolone levels and 5alpha-reductase enzyme activity and expression. Alcohol Clin Exp Res 33:2077–2087

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Titus MA, Li Y, Kozyreva OG, Maher V, Godoy A, Smith GJ, Mohler JL (2013) 5α-reductase type 3 enzyme in benign and malignant prostate. Prostate. doi:10.1002/pros.22745

    PubMed Central  PubMed  Google Scholar 

  • Torres JM, Ortega E (2003a) Alcohol intoxication increases allopregnanolone levels in female adolescent humans. Neuropsychopharmacology 28:1207–1209

    CAS  PubMed  Google Scholar 

  • Torres JM, Ortega E (2003b) Differential regulation of steroid 5alpha-reductase isozymes expression by androgens in the adult rat brain. FASEB J 17:1428–1433

    Article  CAS  PubMed  Google Scholar 

  • Torres JM, Ortega E (2004) Alcohol intoxication increases allopregnanolone levels in male adolescent humans. Psychopharmacology 172:352–355

    Article  CAS  PubMed  Google Scholar 

  • Torres JM, Ortega E (2006) Steroid 5alpha-reductase isozymes in the adult female rat brain: central role of dihydrotestosterone. J Mol Endocrinol 36:239–245

    Article  CAS  PubMed  Google Scholar 

  • VanDoren MJ, Matthews DB, Janis GC, Grobin AC, Devaud LL, Morrow AL (2000) Neuroactive steroid 3α-hydroxy-5α-pregnan-20-one modulates electrophysiological and behavioral actions of ethanol. J Neurosci 20:1982–1989

    CAS  PubMed  Google Scholar 

  • Waszkiewicz N, Szajda SD, Zalewska A, Szulc A, Kępka A, Minarowska A, Wojewódzka-Żelezniakowicz M, Konarzewska B, Chojnowska S, Ladny JR, Zwierz K (2012) Alcohol abuse and glycol conjugate metabolism. Folia Histochem Cytobiol 50:1–11

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was funded in part by the European Fund of Regional Development (FEDER-BFU2008-05340) and the Andalusian Regional Government (Endocrinology and Metabolism Group).

Conflict of interest

The authors do not have direct financial conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Granada, Avda. de Madrid s/n, 18012, Granada, Spain

    Pilar Sánchez, Beatriz Castro, Jesús M. Torres & Esperanza Ortega

  2. Institute of Neurosciences, Faculty of Medicine, University of Granada, Granada, Spain

    Jesús M. Torres & Esperanza Ortega

Authors
  1. Pilar Sánchez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Beatriz Castro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jesús M. Torres
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Esperanza Ortega
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jesús M. Torres or Esperanza Ortega.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sánchez, P., Castro, B., Torres, J.M. et al. Effects of different ethanol-administration regimes on mRNA and protein levels of steroid 5α-reductase isozymes in prefrontal cortex of adolescent male rats. Psychopharmacology 231, 3273–3280 (2014). https://doi.org/10.1007/s00213-014-3558-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00213-014-3558-6

Keywords

Search

Navigation