Early-life exposure to three consecutive and brief isoflurane anesthesias or maternal separations do not produce long-lasting depression-related behavior.

<p>Saccharin preference (<b>A, E</b>). Latency to immobility (<b>B, F</b>) and relative immobility during 2–6 min (<b>C, G</b>) in forced swimming test. Stress-induced hyperthermia (<b>D, H</b>). Abbreviations: ISO, isoflurane; MS, maternal separation; FST, forced swimming test; SIH, stress-induced hyperthermia. N = 6-8/group.</p

Early-life exposure to three consecutive and brief isoflurane anesthesias or maternal separations do not produce long-lasting schizophrenia-related behavior.

<p>Nesting behavior (<b>A, C</b>). Prepulse Inhibition (prepulse 68dB or 80dB) (<b>B, D</b>). Abbreviations: ISO, isoflurane; MS, maternal separation; PPI, prepulse inhibition. N = 6-8/group.</p

Experimental time-line.

<p>At postnatal days 7, 8 and 9 (P7-9) or 15, 16 and 17 (P15-17) male mice were subjected to 30 min maternal separation or 30 min isoflurane anesthesia, or were left unhandled (sham). Animals were weaned at P21 (week 3; W3). Beginning from age of 9-weeks animals were subjected to a behavioral phenotyping battery consisting of circadian activity, nest construction and saccharin preference (InfraMot monitoring system), light-dark box, open field, prepulse inhibition (PPI), water maze, stress-induced hyperthermia and forced swim test. N = 6-8/group.</p

Animals exposed to repeated brief isoflurane anesthesia or maternal separation at postnatal days 15–17 show varying circadian activity at adult age.

<p>Hourly average circadian activity during 7-day monitoring (<b>A, D</b>), average circadian activity during different light-cycles (<b>B, E</b>) and average circadian activity during the first dark period in comparison to remaining dark periods (<b>C, F</b>). Lights off (active period; grey) during 6:00 PM– 6:00 AM. Abbreviations: ISO, isoflurane; MS, maternal separation. ***<0.001 two-way ANOVA followed by Newman-Keuls <i>post hoc</i> test MS vs. Sham. ###<0.001, ##<0.01 two-way ANOVA followed by Newman-Keuls <i>post hoc</i> test ISO vs. MS. N = 6-8/group.</p

Early-life exposure to three consecutive and brief isoflurane anesthesias or maternal separations at postnatal days 7–9 bring mild deficit in spatial navigation memory.

<p>Latency to escape (find the platform) (<b>A, E</b>) and time spent near the vicinity of pre-existing platform (quadrant, zone) (<b>B, F</b>) during the first learning trials and probe test, respectively. Latency to escape (find the platform) (<b>C, G</b>) and time spent near the vicinity of pre-existing platform (quadrant, zone) (<b>D, H</b>) during the reverse learning trials and probe test, respectively. Abbreviations: ISO, isoflurane; MS, maternal separation. *<0.05, **<0.01, two-way ANOVA followed by Newman-Keuls <i>post hoc</i> test. N = 6-8/group.</p

Early-life exposures to three consecutive and brief isoflurane anesthesias at postnatal days 15–17 produce mild long-lasting decrease in anxiety-related behavior.

<p>Distance travelled (and habituation) (<b>A, G</b>) and time in center (<b>B, H</b>) in a novel open environment (open field test). Latency to light (<b>C, I</b>), relative distance travelled (<b>D, J</b>), distance in light (<b>E, K</b>) and rearings in light (<b>F, L</b>) in the light-dark box test. Abbreviations: ISO, isoflurane; MS, maternal separation; OF, open field test; LD, light-dark box test. *<0.05, two-way ANOVA followed by Newman-Keuls <i>post hoc</i> test. N = 6-8/group.</p

Immunohistochemical localization of CD73 in mouse brain.

<p>Coronal sections of WT (panels A, C and E) and CD73 deficient (panels B, D and F) mouse cerebrums stained with CD73 antibody. The specific CD73 staining is detected in caudoputamen (CP), olfactory tubercle (OT), globus pallidus (GP) and meninx (arrow). Scale bar 500 μm.</p

CD73 deficient and wt mice display similar pre-pulse inhibition but startle response is enhanced in CD73 deficient mice compared to wt mice.

<p>Percentage of pre-pulse inhibition averaged across all pre-pulse intensities. Insert bar graphs represent percentage of alteration of startle response without pre-pulse stimulus from the wild type animals’ startle response level (wt mice, n = 38; CD73 deficient mice, n = 29). The results from both sexes are combined, because there was no <i>sex</i> effect or <i>sex</i> X <i>genotype</i> interaction. Mean values are plotted with SEM, **p<0.01 t-test.</p

CD73 controls circadian changes in locomotor activity in isolated mice.

<p>(A) Locomotor activity presented as activity counts during 72 hr observation period, when the mice were separated in individual cages. These data are presented for both sexes together, because there was no <i>sex</i> effect or <i>sex</i> X <i>genotype</i> interaction (CD73 deficient mice, n = 25; wt mice, n = 30). (B) Locomotor activity measured as corner visits in IntelliCage. In IntelliCage analyses only females were used (CD73 deficient, n = 9; wt, n = 8). Mean values are plotted with SEM, # p<0.05 and ## p<0.01, repeated ANOVA.</p

CD73 deficient mice display altered social behaviour.

<p>(A) Percentages of the times pushed out in the tube test (32 mice/group in both genotypes). (B) Percentage of the time spent in social activity towards the intruder in the resident-intruder test (wt mice, n = 15; CD73 deficient mice, n = 13). (C) Time spent in sniffing the stranger in the social novelty preference test, and (D) time spent in sniffing a new and an old stranger in the sociability test (wt, n = 16; CD73 deficient, n = 16). Mean values are plotted with SEM, *p<0.05 t-test, **p<0.01 t-test.</p