1. Block et al., 2012. Are anesthesia and surgery during infancy associated with altered academic performance
during childhood? Anesthesiology, 117:494-503.
Fredriksson et al., 2007. Neonatal exposure to a combination of N-methyl-D-aspartate and gamma-
aminobutyric acid type A receptor anesthetic agents potentiates apoptotic neurodegeneration and persistent
behavioral deficits. Anesthesiology, 107(3):427-36.
Jevtovic-Todorovic et al., 2003. Early Exposure to Common Anesthetic Agents Causes Widespread
Neurodegeneration in the Developing Rat Brain and Persistent Learning Deficits. The Journal of
Neuroscience, 23(3):876 – 882.
Mintz et al., 2013. Anesthetics interfere with axon guidance in developing mouse neocortical neurons in vitro
via a γ-aminobutyric acid type A receptor mechanism. Anesthesiology, (4):825-33.
Palanisamy et. al, 2011. Rats Exposed to Isoflurane In Utero during Early Gestation Are Behaviorally
Abnormal as Adults. Anesthesiology, 114(3):521-28.
Ramage et al., 2013. Distinct long-term neurocognitive outcomes after equipotent sevoflurane or isoflurane
anaesthesia in immature rats. British Journal of Anaesthesia 110 (S1): i39–i46.
Schuckit MA (1996) Recent developments in the pharmacotherapy of alcohol dependence. Journal of
consulting and clinical psychology 64:669-676.
 Anesthetic exposure during critical periods of development is associated with
cognitive and behavioral deficits
 Neuronal:
• Alterations of axonal guidance (Mintz, 2013).
• Evidence of cell death and neurotoxicity (Fredriksson et al., 2007; Jevtovic-Todorovic et al., 2003).
 Behavioral:
• Deficits in short-term, long-term, (Ramage et al., 2013) and spatial memory (Palanisamy et. al,
2011).
• Higher levels of anxiety (Palanisamy et. al, 2011).
• In humans- Decrease in standardized test scores (Block et al., 2012).
 Few studies have examined anesthetic effects during the vulnerable period of
adolescence
 We previously showed that adolescent anesthetic exposure alters sensitivity to
sedative/hypnotic and cognitive effects of alcohol:
• Persistent decreases in sensitivity to the sedative/hypnotic effects of alcohol
• Increased sensitivity to the memory-impairing effects of alcohol
• Alterations may be mediated by isoflurane-induced increases in a4 and d GABAA-Rs
 Importantly, reduction of sensitivity to sedative/hypnotic effects of ethanol is
associated with increased ethanol consumption (Schuckit 1996).
 In the present study, we examined the effects of isoflurane exposure in
adolescents on subsequent ethanol-intake behaviors.
General anesthetic exposure effects on ethanol intake in adolescent rats.
Kristin Eggers, Elizabeth Ventura, Justine D. Landin, David F. Werner
Center for Development and Behavioral Neuroscience, Department of Psychology, Binghamton University, Binghamton, NY 13902
 Isoflurane exposure increases animals’ propensity to drink both the
ethanol and sucrose solutions.
 This may indicate an alteration in appetitive reinforcement, and/or
reward pathways following adolescent isoflurane exposure
 Preliminary data suggests that adolescent isoflurane exposure
may not increase ethanol preference.
 This is likely due to the heightened sucrose consumption
in isoflurane-exposed adolescents.
 Future studies will continue to assess the effect of isoflurane
exposure on subsequent ethanol behaviors, and their associative
neurobiological alterations.
Figure 1. Adolescent anesthetic exposure increases
EtOH and sucrose drinking time one week later
Figure 4. Isoflurane-exposed adolescents do not differ
from air-exposed adolescents on % EtOH preference
Figure 1. Seconds spent with the ethanol bottle (A) and the sucrose bottle (B)
during 30 minute 2-bottle choice sessions were observed in air-exposed and
isoflurane-exposed adolescents. Data is collapsed across all 5 sessions. On
average, adolescent rats exposed to isoflurane spent significantly more time
drinking both the EtOH and sucrose bottles. Data is presented as mean + SEM,
** p < 0.005, *** p < 0.0005, n= 6/group.
Subjects: Experiments were conducted in accordance with the National
Institute of Health Guidelines under Institutional Animal Care and Use
Committee-approved protocols at the State University of New York –
Binghamton. Adolescent (P28) male Sprague-Dawley rats were used. Rats
were purchased from Taconic (Germantown, NY, USA) or bred in house. Rats
were group-housed and handled before testing. Rats were maintained on a
standard 12 h light–dark schedule with ad libitum access to rat chow and
water.
Drugs: The volatile anesthetic isoflurane (3.0%) or air was administered for 40
minutes 1 week prior to start of testing. The 2-bottle choice tests used one
bottle of a a super-saccharin (.3% sucrose, .125% saccharin) solution in water
and one bottle of 10% ethanol solution in the super-saccharin solution.
Ethanol Intake
2-Bottle Choice Test: Bottles were filled with EtOH or super-saccharin
solution and weighed. Rats were were weighed and littermates were
separated using a mesh divider. Rats were moved into the testing room for
acclimation, 15 minutes prior to start of testing. Rats were given a choice of
10% ethanol solution or sucrose for 30 minutes. The drinking session was
filmed for subsequent analysis. After the 30 minute session, bottles were
weighed to determine the amount of each solution the rats consumed and
calculate mL/kg consumption. In total, there were five 30 minute sessions
conducted every other day.
24-Hour Trial: Following five 2-bottle choice test trials, the animals underwent
a 24-hour trial in which they had free access to separate bottle containing 10%
ethanol in super-saccharin solution and water. Bottles and animals were
weighed prior to and immediately following the trial. Animal weight was
determined by averaging weight prior to and after the 24 hour session.
Statistics: All data utilized air-exposed adolescents as controls. All
experiments were analyzed using unpaired one-way t-tests. For multiple t-
tests in figure 2, a Bonferroni correction was used. Seconds spent drinking
was recorded by a trained observer.
References
Acknowledgements
Figure 2. Adolescent anesthetic exposure increases
time spent with ethanol bottles in a 2-bottle choice test
Figure 2. Seconds spent with the ethanol bottle during 30 minute 2-bottle
choice sessions was observed in air-exposed and isoflurane-exposed
adolescents. Data is represented for each session. On days 1, 3, and 5,
isoflurane-exposed adolescents spent more time with ethanol bottles. On day
2, control animals spent more time with ethanol bottles. Data is presented as
mean + SEM, ^ p = 0.052, * p < 0.05, n= 6/group.
Figure 3. EtOH preference was calculated from seconds with ethanol bottle (A)
or mL/kg consumption (B) across 5 drinking sessions (A) and in one 24 hour
session (B), using the formula: EtOH consumption/EtOH and sucrose
consumption * 100. Isoflurane exposure had no effect on preference for EtOH at
either 30 minutes or 24 hours. Data is presented as mean + SEM, n= 6/group.
A
B
A
B 24 hours
This work was supported by the National Institute of Health
grants AA017823 and AA019367.
0
20
40
60
80
100
Days 1-5
Air Iso
%EtOHpreference
Air
TimewithEtOH(sec.)TimewithEtOH(sec.)
TimewithEtOH(sec.)