Summary

适度的产前酒精暴露在成年大鼠社会行为的量化

Published: December 14, 2014
doi:

Summary

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

Abstract

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several independent laboratories have demonstrated robust alterations in the social behavior of rodents exposed to alcohol during brain development across a wide range of exposure durations, timing, doses, and ages at the time of behavioral quantification. Prior work from this laboratory has identified reliable alterations in specific forms of social interaction following moderate prenatal alcohol exposure (PAE) in the rat that persist well into adulthood, including increased wrestling and decreased investigation. These behavioral alterations have been useful in identifying neural circuits altered by moderate PAE1, and may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior. This paper describes procedures for performing moderate PAE in which rat dams voluntarily consume ethanol or saccharin (control) throughout gestation, and measurement of social behaviors in adult offspring.

Introduction

儿童估计有1-5%被诊断患有胎儿酒精谱系障碍(FASDs)2,其中包括胎儿酒精综合症(FAS),部分FAS(疫区),与酒精相关的神经发育障碍(ARNDs)3。在社会行为和认知赤字之间的儿童FASDs 4-7观察到的最常见的不良后果。消极后果不限于重产前酒精暴露(PAE),中度PAE不导致FAS的显眼形态,行为和认知缺陷特性可引起比较微妙,但仍然持续存在,缺陷在人类与FASDs 8-10和大脑发育11在非人类的动物暴露于乙醇。理解适度PAE的行为和相应的神经生物学后果的重要性是由这表明绝大多数的胎儿酒精中毒综合症病例属于少秒内目前的估计强调艾威范围频谱12。

几个独立的实验室报告了啮齿类动物的社会行为改变大脑发育过程中乙醇暴露相关,包括减少调查和互动1,13-15,改变打法14,16,17,增加响应积极的互动17,18,改建为社会刺激19-21,和赤字在社会上获得的食物的喜好和社会的认可存储器22。社会行为缺陷已观察暴露于重(血液乙醇浓度(BEC中)〜300毫克/分升)22,23或乙醇更温和的水平(BEC中〜80毫克/分升)1,并在广泛的范围内的其他显著参数因素,包括曝光定时,曝光的持续时间,并在行为测定时的年龄。

以前的研究已经证明,改变社会INT的具体方面eraction在成年大鼠歧视暴露在中等水平的酒精从控制动物暴露于糖精1,18。特别是,适度PAE一直被与强劲增长的摔跤,这表明增加的攻击行为,并在成年后社会调查( 例如 ,嗅探合作伙伴)水平较低有关。因为在社会行为改变是PAE的可靠的后果,以下的PAE社会行为的量化可以保持为前进朝向PAE相关的改变中的社会行为的神经基础的更完整的理解和介入方法的发展的重要性。本文和相关视频的目标是提供指令的温和PAE协议和方法,用于在成年子女社会行为具有的量化可靠地分辨产前醇暴露从非暴露鼠后代。

Protocol

在这里和在所附的视频中描述的所有程序已经批准的健康科学中心的机构动物护理和使用委员会和新墨西哥大学的主校区。 1.产前乙醇暴露获得所有需要的材料和化工产品:糖精钠盐水合物,190证明乙醇(95%的酒精体积比),玻璃珠饮水管,天然橡胶白色#4塞一个孔,1个“弯管球点,纸统治者从WWW印刷.vendian.org / mncharity / DIR3 / paper_rulers /。 获得成熟的成年?…

Representative Results

以上的许多育种轮的过程中雌性大鼠的乙醇条件一致喝平均约2.1克/公斤的每4小时饮用会话乙醇。鼠水坝引入饮用管后的第一个15至30分钟期间消耗大约一半的四个小时的总的,产生了峰值母体血清乙醇浓度为约60毫克/分升,在45分钟的时间点测量。在剩余3.5小时的饮用期间,他们继续消耗5%乙醇0.4克/公斤体重/小时的低,但相对稳定的速率。自愿乙醇消费的这种水平和模式,对孕妇体重增加,后?…

Discussion

此处所描述的产前酒精暴露范例涉及在妊娠期间的乙醇自愿食用(5%V / V)的大鼠水坝。有许多用于在文献中,这相对于乙醇给药时间,剂量,时间和路线,以及所研究的物种不同表示脑发育过程中暴露的非人类的动物到乙醇的协议。虽然这里不提供彻底治疗各种暴露协议的优点,自愿饮用方法PAE的几个优点在本协议中描述的高亮显示。以前我们使用的流质饮食协议,常用于研究这方面的做法,?…

Disclosures

The authors have nothing to disclose.

Acknowledgements

Support provided by grant AA019462 to DAH and AA019884 to DDS.

Materials

Saccharin sodium salt hydrate Sigma S1002
190 proof ethanol Sigma 493538
Beaded glass drinking tubes Fisher 14-955K
Natural rubber white #4 stopper one hole Plasticoid LSG4M181
1" bend tubes-ball point Ancare TD-199-3"
Paper rulers N/A N/A www.vendian.org/mncharity/dir3/paper_rulers
Apparatus for social interaction Custom built N/A 95 cm X 47 cm X 43 cm
Video cameras N/A N/A Capable of recording low/no light conditions
Infrared illuminators Vitek VT-IR1-12
Teklad laboratory grade sani-chips Harlan 7090A
Brush and dustpan N/A N/A
Isopropyl alcohol Sigma W292907
Chlorine Dioxide (1.5 mg Tablets) Quiplabs N/A Prepare per manufacturer's recommendation

References

  1. Hamilton, D. A., et al. Prenatal exposure to moderate levels of ethanol alters social behavior in adult rats: Relationship to structural plasticity and immediate early gene expression in frontal cortex. Behav. Brain Res. 207 (2), 290-304 (2010).
  2. May, P. A., et al. Approaching the prevalence of the full spectrum of Fetal Alcohol Spectrum Disorders in a South African population-based study. Alcohol. Clin. Exp. Res. 37 (5), 818-830 (2013).
  3. Chasnoff, I. J., Wells, A. M., Telford, E., Schmidt, C., Messer, G. Neurodevelopmental functioning in children with FAS, pFAS, and ARND. Journal of Developmental and Behavioral Pediatrics. 31 (3), 192-201 (2010).
  4. Disney, E. R., Iacono, W., McGue, M., Tully, E., Legrand, L. Strengthening the case: Prenatal alcohol exposure is associated with increased risk for conduct disorder. Pediatrics. 122 (6), E1225-E1230 (2008).
  5. Kelly, S. J., Goodlett, C. R., Hannigan, J. H. Animal models of fetal alcohol spectrum disorders: Impact of the social environment. Dev Disabil Res Rev. 15 (3), 200-208 (2009).
  6. Kelly, S. J., Day, N., Streissguth, A. P. Effects of prenatal alcohol exposure on social behavior in humans and other species. Neurotoxicol. Teratol. 22 (2), 143-149 (2000).
  7. Thomas, S. E., Kelly, S. J., Mattson, S. N., Riley, E. P. Comparison of social abilities of children with fetal alcohol syndrome to those of children with similar IQ scores and normal controls. Alcoholism: Clinical and Experimental Research. 22 (2), 528-533 (1998).
  8. Conry, J. Neuropsychological deficits in Fetal Alcohol Syndrome and fetal alcohol effects. Alcohol. Clin. Exp. Res. 14 (5), 650-655 (1990).
  9. Streissguth, A. P., et al. Fetal Alcohol Syndrome in adolescents and adults. JAMA-Journal of the American Medical Association. 265 (15), 1961-1967 (1991).
  10. Streissguth, A. P., Barr, H. M., Sampson, P. D. Moderate prenatal alcohol exposure: effects on child IQ and learning problems at age 7 1/2 years. Alcoholism: Clinical and Experimental Research. 14 (5), 662-669 (1990).
  11. Valenzuela, C. F., Morton, R. A., Diaz, M. R., Topper, L. Does moderate drinking harm the fetal brain? Insights from animal models. Trends Neurosci. 35 (5), 284-292 (2012).
  12. May, P. A., et al. Prevalence of children with severe Fetal Alcohol Spectrum Disorders in communities near Rome, Italy: New estimated rates are higher than previous estimates. Int. J. Env. Res. Public Health. 8 (6), 2331-2351 (2011).
  13. Tunc-Ozcan, E., Ullmann, T. M., Shukla, P. K., Redei, E. E. Low-dose thyroxine attenuates autism-associated adverse affects of fetal alcohol in male offspring’s social behavior and hippocampal gene expression. Alcohol. Clin. Exp. Res. 37 (11), 1986-1995 (2013).
  14. Middleton, F. A., Varlinskaya, E. I., Mooney, S. M. Molecular substrates of social avoidance seen following prenatal ethanol exposure and its reversal by social enrichment. Dev. Neurosci. 34 (2-3), 115-128 (2012).
  15. Varlinskaya, E. I., Mooney, S. M. Acute exposure to ethanol on gestational day 15 affects social motivation of female offspring. Behav. Brain Res. 261, 106-109 (2014).
  16. Meyer, L. S., Riley, E. P. Social play in juvenile rats prenatally exposed to alcohol. Teratology. 34 (1), 1-7 (1986).
  17. Royalty, J. Effects of prenatal ethanol exposure on juvenile play-fighting and postpubertal aggression in rats. Psychol Rep. 66 (2), 551-560 (1990).
  18. Hamilton, D. A., et al. Effects of moderate prenatal ethanol exposure and age on social behavior, spatial response perseveration errors and motor behavior. Behav. Brain Res. 269, 44-54 (2014).
  19. Kelly, S. J., Dillingham, R. R. Sexually dimorphic effects of perinatal alcohol exposure on social interactions and amygdala DNA and DOPAC concentrations. Neurotoxicol. Teratol. 16 (4), 377-384 (1994).
  20. Lugo, J. N., Marino, M. D., Cronise, K., Kelly, S. J. Effects of alcohol exposure during development on social behavior in rats. Physiology and Behavior. 78 (2), 185-194 (2003).
  21. Lugo, J. N., Marino, M. D., Gass, J. T., Wilson, M. A., Kelly, S. J. Ethanol exposure during development reduces resident aggression and testosterone in rats. Physiology and Behavior. 87 (2), 330-337 (2006).
  22. Kelly, S. J., Tran, T. D. Alcohol exposure during development alters social recognition and social communication in rats. Neurotoxicol. Teratol. 19 (5), 383-389 (1997).
  23. Mooney, S. M., Varlinskaya, E. I. Acute prenatal exposure to ethanol and social behavior: Effects of age, sex, and timing of exposure. Behav. Brain Res. 216 (1), 358-364 (2011).
  24. Savage, D. D., Becher, M., de la Torre, A. J., Sutherland, R. J. Dose-dependent effects of prenatal ethanol exposure on synaptic plasticity and learning in mature offspring. Alcohol. Clin. Exp. Res. 26 (11), 1752-1758 (2002).
  25. Rasmussen, D. D., et al. Chronic daily ethanol and withdrawal: 1. Long-term changes in the hypothalamo-pituitary-adrenal axis. Alcohol. Clin. Exp. Res. 24 (12), 1836-1849 (2000).
  26. Savage, D. D., et al. Effects of a Novel Cognition-Enhancing Agent on Fetal Ethanol-Induced Learning Deficits. Alcohol. Clin. Exp. Res. 34 (10), 1793-1802 (2010).
  27. Goodlett, C. R., Johnson, T. B. Neonatal binge ethanol exposure using intubation: Timing and dose effects on place learning. Neurotoxicol. Teratol. 19 (6), 435-446 (1997).
  28. Staples, M. C., Rosenberg, M. J., Allen, N. A., Porch, M. W., Savage, D. D. . Impact of Combined Prenatal Ethanol and Prenatal Stress Exposure on Anxiety and Hippocampal-Sensitive Learning in Adult. 37 (12), 2039-2047 (2013).
  29. Champagne, F. A., Francis, D. D., Mar, A., Meaney, M. J. Variations in maternal care in the rat as a mediating influence for the effects of environment on development. Physiol. Behav. 79 (3), 359-371 (2003).
  30. Hamilton, D. A., et al. Patterns of social-experience-related c-fos and Arc expression in the frontal cortices of rats exposed to saccharin or moderate levels of ethanol during prenatal brain development. Behav. Brain Res. 214 (1), 66-74 (2010).
  31. Barnett, S. A. . A study in behaviour: Principles of ethology and behavioural physiology displayed mainly in the rat. , (1963).
  32. Pellis, S. M., Pellis, V. C. Play-fighting differs from serious fighting in both target of attack and tactics of fighting in the laboratory rat Rattus-Norvegicus. Aggressive Behav. 13 (4), 227-242 (1987).
  33. Meaney, M. J., Stewart, J. A descriptive study of social-development in the rat (Rattus-Norvegicus). Anim. Behav. 29 (1), 34-45 (1981).
  34. Pellis, S. M., Pellis, V. C. The prejuvenile onset of play fighting in laboratory rats (Rattus norvegicus). Dev. Psychobiol. 31 (3), 193-205 (1997).
  35. Himmler, B. T., Pellis, V. C., Pellis, S. M. Peering into the dynamics of social interactions: measuring play fighting in rats. J. Vis. Exp. (71), e4288-e4288 (2013).
  36. Panksepp, J., Siviy, S., Normansell, L. The psychobiology of play : Theoretical and methodological perspectives. Neurosci. Biobehav. Rev. 8 (4), 465-492 (1984).
  37. Siviy, S. M., Panksepp, J. Sensory modulation of juvenile play in rats. Dev. Psychobiol. 20 (1), 39-55 (1987).
  38. Pellis, S. M., et al. The effects of orbital frontal cortex damage on the modulation of defensive responses by rats in playful and nonplayful social contexts. Behav. Neurosci. 120 (1), 72-84 (2006).
  39. Blanchard, R. J., Blanchard, D. C., Takahashi, T., Kelley, M. J. Attack and defensive behavior in albino-rat. Anim. Behav. 25 (3), 622-634 (1977).

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Cite This Article
Hamilton, D. A., Magcalas, C. M., Barto, D., Bird, C. W., Rodriguez, C. I., Fink, B. C., Pellis, S. M., Davies, S., Savage, D. D. Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats. J. Vis. Exp. (94), e52407, doi:10.3791/52407 (2014).

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