Name: moderate prenatal alcohol exposure and quantification of social behavior in adult rats
Uploaded: 2014-12-14T14:00:00
Description: Watch this Scientific Journal Video about Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats at JoVE.com

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

All procedures described here and in the accompanying video have been approved by the Institutional Animal Care and Use Committees of the Health Sciences Center and the main campus of the University of New Mexico.
1. Prenatal Ethanol Exposure
<ol>
	<li>Obtain all required materials and chemicals: Saccharin sodium salt hydrate, 190 proof ethanol (95% alcohol by volume), beaded glass drinking tubes , natural rubber white #4 stopper one hole , 1&#34; bend tubes-ball point, paper rulers printed ...

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

The prenatal alcohol exposure paradigm described here involves voluntary consumption of ethanol (5% v/v) by rat dams during pregnancy. There are a number of protocols for exposing non-human animals to ethanol during brain development represented in the literature, which differ with respect to the timing, dose, duration and route of ethanol administration as well as the species under investigation. Although a thorough treatment of the advantages of various exposure protocols is not provided here, several advantages of the...

An estimated 1-5% of children are diagnosed with Fetal Alcohol Spectrum Disorders (FASDs)2, which include Fetal Alcohol Syndrome (FAS), partial FAS (pFAS), and Alcohol-Related Neurodevelopmental Disorders (ARNDs)3. Deficits in social behavior and cognition are among the most common adverse outcomes observed in children with FASDs4-7. Negative consequences are not limited to heavy prenatal alcohol exposure (PAE), as moderate PAE that does not lead to the conspicuous morphological, behavioral and cognitive deficits characteristic of FAS can cause comparatively subtle, but nonetheless persistent, deficits in humans with FASDs8-10 and non-human animals exposed to ethanol during brain development11. The importance of understanding the behavioral and corresponding neurobiological consequences of moderate PAE is underscored by current estimates indicating that the large majority of FASD cases fall within the less severe range of the spectrum12.
Several independent laboratories have reported alterations in rodent social behavior related to ethanol exposure during brain development, including decreased investigation and interaction1,13-15, altered play14,16,17, increased aggressive interactions17,18, alterations in responsiveness to social stimuli19-21, and deficits in socially acquired food preferences and social recognition memory22. Social behavior deficits have been observed following exposure to heavy (blood ethanol concentrations (BECs) ~300mg/dl)22,23 or more moderate levels of ethanol (BECs ~80mg/dl)1, and across a broad range of parameters for other significant factors including exposure timing, duration of exposure, and age at the time of behavioral measurement.
Previous research has demonstrated that alterations in specific aspects of social interaction in adulthood discriminate rats exposed to moderate levels of alcohol from control animals exposed to saccharin1,18. In particular, moderate PAE has consistently been associated with robust increases in wrestling, which suggests increases in aggressive behavior, and lower levels of social investigation (e.g., sniffing of the partner) in adulthood. Because alterations in social behavior are reliable consequences of PAE, the quantification of social behavior following PAE may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior and the development of interventional approaches. The goal of this paper and the associated video is to provide instruction on the moderate PAE protocol and methods for quantification of social behavior in adult offspring that have reliably distinguished prenatal alcohol-exposed from non-exposed rat offspring.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Saccharin sodium salt hydrate</td>
			<td>Sigma</td>
			<td>S1002</td>
			<td></td>
		</tr>
		<tr>
			<td>190 proof ethanol</td>
			<td>Sigma</td>
			<td>493538</td>
			<td></td>
		</tr>
		<tr>
			<td>Beaded glass drinking tubes</td>
			<td>Fisher</td>
			<td>14-955K</td>
			<td></td>
		</tr>
		<tr>
			<td>Natural rubber white #4 stopper one hole</td>
			<td>Plasticoid</td>
			<td>LSG4M181</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#34; bend tubes-ball point</td>
			<td>Ancare</td>
			<td>TD-199-3&#34;</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper rulers</td>
			<td>N/A</td>
			<td>N/A</td>
			<td><a href="http://www.vendian.org/mncharity/dir3/paper_rulers">www.vendian.org/mncharity/dir3/paper_rulers</a></td>
		</tr>
		<tr>
			<td>Apparatus for social interaction</td>
			<td>Custom built</td>
			<td>N/A</td>
			<td>95 cm X 47 cm X 43 cm</td>
		</tr>
		<tr>
			<td>Video cameras</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Capable of recording low/no light conditions</td>
		</tr>
		<tr>
			<td>Infrared illuminators</td>
			<td>Vitek</td>
			<td>VT-IR1-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Teklad laboratory grade sani-chips</td>
			<td>Harlan</td>
			<td>7090A</td>
			<td></td>
		</tr>
		<tr>
			<td>Brush and dustpan</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl alcohol</td>
			<td>Sigma</td>
			<td>W292907</td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorine Dioxide (1.5 mg Tablets)</td>
			<td>Quiplabs</td>
			<td>N/A</td>
			<td>Prepare per manufacturer&#39;s recommendation</td>
		</tr>
	</tbody>
</table>

moderate prenatal alcohol exposure and quantification of social behavior in adult rats

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.

Over the course of many breeding rounds female rats in the ethanol condition consistently drink an average of about 2.1 g/kg of ethanol per 4 hr drinking session. Rat dams consume approximately one-half of the four hr total during the first 15 to 30 min after the introduction of the drinking tubes, resulting in a peak maternal serum ethanol concentration of about 60 mg/dl, measured at the 45 min time point. Over the remaining 3.5 hr of the drinking period, they continue to consume 5% ethanol at a lower, but relatively st...

Watch this Scientific Journal Video about Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats at JoVE.com

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

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.

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several ...

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Behavior

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

A Method for Evaluating the Reinforcing Properties of Ethanol in Rats without Water Deprivation, Saccharin Fading or Extended Access Training

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods require saccharin/sucrose fading, water deprivation and/or extended training to initiate operant responding in rats. This paper describes a novel and efficient method to quickly initiate operant responding for ethanol that is convenient for experimenters and does not require water deprivation or saccharin/sucrose fading, thus eliminating the potential confound of using sweeteners in ethanol operant self-administration studies. With this method, Wistar rats typically acquire and maintain self-administration of a 20% ethanol solution in less than two weeks of training. Furthermore, blood ethanol concentrations and rewards are positively correlated for a 30 min self-administration session. Moreover, naltrexone, an FDA-approved medication for alcohol dependence that has been shown to suppress ethanol self-administration in rodents, dose-dependently decreases alcohol intake and motivation to consume alcohol for rats self-administering 20% ethanol, thus validating the use of this new method to study the reinforcing properties of alcohol in rats.

This protocol describes a novel and efficient method to quickly initiate operant responding for ethanol in rats that, contrary to standard methods, does not require water deprivation or saccharin/sucrose fading to initiate responding.

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods ...

Examining Recall Memory in Infancy and Early Childhood Using the Elicited Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is commonly assessed using verbal report paradigms in adults, studying the development of this ability in preverbal infants and children proved challenging. Over the past 30 years, researchers have developed a non-verbal means of assessing recall memory known as the elicited or deferred imitation paradigm. In one variant of the procedure, participants are presented with novel three-dimensional stimuli for a brief baseline period before a researcher demonstrates a series of actions that culminate in an end- or goal-state. The participant is allowed to imitate the demonstrated actions immediately, after a delay, or both. Recall performance is then compared to baseline or to performance on novel control sequences presented at the same session; memory can be assessed for the individual target actions and the order in which they were completed. This procedure is an accepted analogue to the verbal report techniques used with adults, and it has served to establish a solid foundation of the nature of recall memory in infancy and early childhood. In addition, the elicited or deferred imitation procedure has been modified and adapted to answer questions relevant to other aspects of cognitive functioning. The broad utility and application of imitation paradigms is discussed, along with limitations of the approach and directions for future research.

The elicited imitation procedure was established to examine the development of recall memory in infancy and early childhood. This procedure has been widely used to establish a solid foundation of the nature of recall memory in infancy and early childhood.

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on well-being and can affect the comparability and reproducibility of research results, the amount of stress experienced by laboratory animals should be minimized and avoided when possible. Further, the direct contact between the rat and animal caretaker during the cage change bears hygiene risks and therefore possibly negatively impacts the well-being of the rats and the quality of the research. 
Our protocol aims to improve the routinely performed cage changing procedure. For this reason, we present a feasible protocol that enables rats to learn via clicker training and observation to voluntarily change to a clean cage. This training helps to reduce stress caused by the physical disturbance and handling associated with the cage changes and concurrently enables a reduction in direct contact between animal and animal caretaker after the training phase is completed.
The implementation of clicker training to rats is fast and easy. Rats are generally interested in the training and efficiently learn the desired behavior, which entails changing cages through a pipe. Even without training, the rats learn to perform the desired behavior by observation, as 80% of the observational learning group successfully changed cages when tested. The training further helps to establish a relationship of trust between trainer and animal. As hygiene and well-being are both very important in animal experiments, this protocol might also help to improve high-quality research.

This protocol introduces a method of cage change for rats via clicker training. Rats learn the desired behavior not only by direct training but also by observational learning. The implementation of this fast and easy protocol might help to improve well-being and hygiene in rodent facilities.

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on ...

Integrating Computerized Linguistic and Social Network Analyses to Capture Addiction Recovery Capital in an Online Community

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery capital development in online communities of recovery from alcohol and drug (AOD) addiction. Recovery capital was conceptualised as both engagement in the online recovery community and identification with the community. To measure recovery capital development, naturally occurring data were extracted from the social media page of a specific recovery program, with the page being set up as a resource for a face-to-face recovery program. To map engagement with the online community, social network analysis (SNA) capturing online social interaction was performed. Social interaction was measured through the linkages between the online contributors/members of the online community as represented by program clients, staff, and supporters from the broader community. To capture markers of social identification with the online community, computerised linguistic analysis of the textual data (content from posts and comments) was conducted. Recovery capital captured in this way was analysed against retention data (a proxy outcome indicator), as days spent in the (face-to-face) recovery program. The online data extracted was linked to participant data in regards to program retention to test prediction of a key recovery outcome. This approach allowed the examination of the role of online support communities and assessment of the association between recovery capital (developed via the online community of recovery) and recovery outcomes.

The article describes a novel approach for analyzing dynamic online social interactions (in an online context) exemplified by a study of an online community of recovery from alcohol and drug addiction.

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery ...

Impact of High-intensity Interval Exercise and Moderate-Intensity Continuous Exercise on the Cardiac Troponin T Level at an Early Stage of Training

An elevation in cardiac troponin T (cTnT), as a highly specific biomarker of cardiomyocyte damage, after moderate-intensity continuous exercise (MCE) has been described. The exercise-induced cTnT response distorts the diagnostic role of the cTnT assay. Although high-intensity interval exercise (HIE) is growing in popularity and concerns remain about its safety, available data related to cTnT release after HIE is limited, which hampers the use of HIE as a health intervention. Here, we present three representative HIE protocols [traditional HIE (repeated 4 min cycling at 90% V&#775;O2max interspersed with 3 min rest, 200 kJ/session); sprint interval exercise (SIE, repeated 1 min cycling at 120% V&#775;O2max interspersed with 1.5 min rest, 200 kJ/session); and repeated sprint exercise (RSE, 40 x 6 s all-out sprints interspersed with 9 s rest)] and one representative MCE protocol (continuous cycling exercise at an intensity of 60% V&#775;O2max, 200 kJ/session). Forty-seven sedentary, overweight young women were randomly assigned to one of four groups (HIE, SIE, RSE, and MCE). Six bouts of respective exercise were performed by every single group, with each being 48 h apart. Meanwhile, for four groups, the duration of the entire testing period was identical, being 10 days. Before and after the first and final exercise bouts, an assessment was conducted of cTnT. The current study provides a frame of reference giving a clear picture of how a specific exercise session affects the circulating cTnT concentration at the early stage of training. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise, especially for HIE.

Here, we present protocols of high-intensity interval and moderate-intensity continuous exercise to observe the response of circulating cardiac troponin T (cTnT) concentration to acute exercise over 10 days. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise.

An elevation in cardiac troponin T (cTnT), as a highly specific biomarker of cardiomyocyte damage, after moderate-intensity continuous exercise (MCE) has ...