Can Mice Make Friends? Social Behavior of Phenotypically Autistic Mice Samantha Giordano a , Sarah Guariglia b , Bradley Rehnberg a , and Guang Wen b a Department of Biological Science, York College of Pennsylvania b Institute of Basic Research, College of Staten Island Introduction ATSDR Report on Brick Township NJ showed higher than EPA standard levels of Bromoform, Chloroform and Tetrachloroethylene were in the drinking water. Autism Rates in Brick Township NJ ATSDR, 1998: 4.0 cases per 1,000 children Bertrand et al., 2001: 6.7 cases per 1,000 children Research found that the triad of chemicals found in Brick Township can cause an increase in the c-AMP RII subunit in the brain in clam embryos (Kreiling et al., 2005). Brain synapse functioning is disrupted in autism Guariglia tested the triad of chemicals on mice. Two of the 3 features of autism tested: 1)Verbal impairments 2)Cognitive/behavioral impairments 3)NO test of Social Impairments*** Question Do the mice treated with the triad of chemicals from Brick Township, NJ show social impairments?? Methods Female mice and their pups were given drinking water with different concentrations, control (0X), 1X, 10X, and 100X, of the triad of chemicals found in Brick Township NJ. Social Behavior Apparatus adapted from Moy et al. (2004) Social Behavior Test 1) Habituation: Experimental mouse was alone in the test apparatus 2) Socialization: Novel mouse 1 (same sex as the experimental mouse) was randomly placed into the left or right cup. The test mouse then entered the test apparatus. 3) Preference for Social Novelty: Novel mouse 2 (same sex as the Experimental Mouse) was placed into the empty cup. Both cups now contained control mice and the test mouse entered the test apparatus. Analysis • Entries into left and right chambers • Time in left and right chambers • Time spent with novel mouse 1 • Time spent with novel mouse 2 • Time spent sniffing novel mouse 1 • Time spent sniffing novel mouse 2 Literature Cited 1)Agency for Toxic Substances and Disease Registry. 2000. Brick township investigation. ATSDR. Available from: http :// www.atsdr.cdc.gov/hac/pha/brick/bti_p1.html 2) Bertrand, J., Mars, A., Boyle, C., Bove, F., Yeargin-Allsopp, M., and Decoufle, P. 2001. Prevalence of Autism in a Untied States Population: Brick Township, New Jersey, Investigation. Pediatrics 108: 1155-1161. 3) Kreiling, J.A., Stephens, R. E., Reinisch, C. L. 2005. A Mixture of environmental contaminants increase c- AMP-dependent protein kinase in Spisula embryos. Environmental Toxicology and Pharmacology 19: 9-18. 4) Moy, S. S., Nadler, J. J., Perez, A., Barbaro, R. P., Johns, J. M., Magnuson, T. R., Piven, J., and Crawley, J.N. 2004. Sociability and preference for social novelty in five inbred strains: an approach to assess autistic- like behavior in mice. Genes, Brain, and Behavior 3: 287-302. Acknowledgements A special thank you to the Institute of Basic Research and the College of Staten Island. 6.4c m 6.4c m 50.8cm 17.1cm ” 17.1cm 16.4cm 24.8cm Divider height: 21cm Doorway height: 6.4cm Behavioral Apparatus Results Conclusions 1) Both male and female mice were on average more interested in novel mouse 2 which is inconsistent with an autistic phenotype. 2) Both male and female mice showed a preference to be social during socialization. 3) Female mice are more social than male mice. 4) A larger sample size was needed to reduce variability. Key Points Figures 1 and 2: Most of the mice preferred being in the area of novel mouse 1. The mice disliked being alone. Key points Figure 2: The male mice, except for the 10X group, preferred novel mouse 2. The 10X group showed a more autistic behavior, a preference for the mouse they know better. Key Points Figure 3: The female treated mice showed behavior opposite the control mice (although not with significant data). The treated mice preferred novel mouse 2 while the control mice preferred novel mouse 1. Key Points Figure 5: The male mice showed a mixture of results; the 1X and 100X groups sniffed novel mouse 1 more than the control while the 10X group did not. All of the female experimental mice sniffed novel mouse 1 less than the control mice. Key points Figure 6: The male groups showed no sniff preference among novel mouse 1 and 2! Key points Figure 7: No definite conclusions could be drawn from the female mice sniff behavior. Fem ale Preference forSocial N ovelty Tim es Original Mouse No vel Mouse Original Mou se Novel Mouse Or iginal Mouse No vel Mouse Original Mou se No vel Mouse 0 100 200 300 400 C ontrol 100X 10X 1X Figure 4.The m ean tim e (sec)spentwith novel m ouse 1 and novel m ouse 2 for the m ale C ontrol,1X,10X,and 100X groups. Errorbars representone standard errorfrom the m ean and an asterisk denotes significance (p<0.05). Unpaired t-tests showed no difference in tim e spentwith novel m ouse 1 and novel m ouse 2 in the control,1X,and 100X groups,p=0.8289,p=0.1605 and p=0.7209, respectively. There was a significantdifference between tim e spentwith novel m ouse 1 and novel m ouse 2 in the 10X group,p=0.0040. * Tim e (seconds) Novel Mouse 1 Novel Mouse 2 Novel Mouse 1 Novel Mouse 2 Novel Mouse 1 Novel Mouse 2 Novel Mouse 1 Novel Mouse 2 0 100 200 300 400 C ontrol 1X 10X 100X M ale Preference forSocial N ovelty Tim es Figure 3. The m ean tim e (sec)spentwith novel m ouse 1 and novel m ouse 2 for the m ale C ontrol,1X,10X,and 100X groups. Errorbars representone standard errorfrom the m ean and the asterisk denotes significance (p<0.05). Unpaired t- tests showed no difference in tim e spentwith novel m ouse 1 and novel m ouse 2 in the control,1X,and 100X groups,p=0.1750,p=0.7081,and p=0.2701, respectively. There was a significantdifference between tim e spentwith novel m ouse 1 and novel m ouse 2 in the 10X group,p=0.0032. * Tim e (seconds) Fem ale Preference forSocial N ovelty SniffTim es 0 20 40 60 80 C ontrol 1X 10X 100X Figure 7. The m ean tim e (sec)spentsniffing novel m ouse 1 and novel m ouse 2 of m ale m ice in C ontrol,1X,10X,100X groups. Errorbars representone standard errorfrom the m ean and astericks denotes signifcantdifferences (p<0.05). Unpaired t-tests showed no significantdifferences between the snifftim e ofnovel m ouse 1 and novel m ouse 2 forthe C ontrol,10X,and 100X,p=0.8475,p=0.3823and p=0.5328, respectively. There was a significantdifference between the 1X sniffing tim e ofnovel m ouse 1 and novel m ouse 2,p=0.0207. * Tim e (seconds) Left Chamber Right Chamber Left Chamber Right Chamber Novel Mouse 1 Novel Mouse 2 M ale Preference forSocial N ovelty SniffTim es N ovelM ouse 1 N ovelM ouse 2 N ovelM ouse 1 N ovelM ouse 2 N ovelM ouse 1 N ovelM ouse 2 N ovelM ouse 1 N ovelM ouse 2 0 50 100 150 200 C ontrol 10X 100X 1X Figure 6. The m ean tim e (sec)spentsniffing novel m ouse 1 and novel m ouse 2 ofm ale m ice in C ontrol,1X,10X,100X groups. Errorbars representone standard errorfrom the m ean. Unpaired t-tests showed no significantdifferences (p<0.05)between the snifftim e ofnovel m ouse 1 and novel m ouse 2 forthe C ontrol,1X,10X,and 100X,p=0.6556, p=0.2864.p=0.5264,and p=0.4515,respectively. Tim e (seconds) Fem ale Socialization Tim es Novel Mouse Empty chamber Nov el Mouse Emp ty Chamber Nov el Mouse Empty Chamber Novel Mouse Empty Chamber 0 100 200 300 400 1X 10X 100X C ontrol Figure 2. M ean tim es (sec)spentwith the novel m ouse vs the em pty cham ber ofthe fem ale m ouse during Socialization in C ontrol,1X,10X,and 100X groups. Errorbars representone standard errorfrom the m ean and the asterisks denote a significantdifference between the groups (p<0.05). Unpaired t-tests showed the p values forthe control,1X,10X,and 100X were p=0.0122,p=0.0239,p=0.0131,and p<0.0001,respectively. * * * * Tim e (seconds) N ovelm ouse Em pty C ham ber N ovel M ouse Em pty C h am ber N ovelM ouse Em pty C h am ber N ovel M ouse Em pty C h am ber 0 100 200 300 400 1X 10X 100X C ontrol M ale Socialization Tim es Figure 1. M ean tim es (sec)spentwith the novel m ouse vs the em pty cham berof the m ale m ouse during Socialization in C ontrol,1X,10X,and 100X groups. Errorbars representone standard errorfrom the m ean and the asterisks denote a significantdifference between the groups (p<0.05). Upaired t-tests showed the p values forthe C ontrol,1X,10X,and 100X are p=0.9866,p=0.0163,p=0.0276, and p=0.0020,respectively. * * * Tim e (seconds) C ontrol 1X 10X 100X C ontrol 1X 10X 100X 0 20 40 60 80 Female Male M ale and Fem ale Socialization SniffTim es Figure 5. The m ean tim es (sec)spentsniffing novel m ouse 1 forboth the m ale and fem ale m ice groups. Errorbars representone standard errorfrom the m ean.ANO VA tests showed no significantdifference (p<0.05)between the control group and the m ale experim ental groups p=0.5400 and the control and fem ale experim ental groups p=0.1366. Tim e (seconds)