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Results: 9

2.
Figure 3

Figure 3. MALTT mice show altered pup ultrasonic vocalization patterns. From: Multiple autism-like behaviors in a novel transgenic mouse model.

Pup ultrasonic vocalizations (USVs), when separated from mother and littermates, were assessed on PND 3–14. (a) Male pup USV development curve is shown, WT n = 21; TG n = 24. (b) Female pup USV development curve is shown, WT n = 19; TG n = 22. All data are presented ± SEM.

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
3.
Figure 2

Figure 2. MALTT mice are hyperactive and exhibit a rotational stereotypy. From: Multiple autism-like behaviors in a novel transgenic mouse model.

Total distance was measured during 10 minutes of open field exploration for mice aged 17, 22, 28, and 45 days. (a) Male total distance is shown. (b) Stereotypies in males were measured as total revolutions/total distance traveled for male mice. (c) Average absolute numbers of revolutions for males on PND45 are shown. (d) Female total distance is shown. All data are presented ± SEM. Males: WT n = 9–17, TG n = 9–13; females: WT n = 12–17, TG n = 11–17; * p ≤ 0.05, ** p ≤ 0.01

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
4.
Figure 6

Figure 6. Juvenile MALTT mice show minimal vocalization in a social pair. From: Multiple autism-like behaviors in a novel transgenic mouse model.

Direct social interaction task two: MALTT + MALTT partner pairs or WT + WT partner pairs were analyzed for USVs during direct social interaction; MALTT n = 9 pairs; WT n = 8 pairs. (a) Mean total USVs emitted from same genotype pairs is shown. Individual pair data is indicated (black circles), with the exception of TG:TG data point “0” which includes 7 pairs’ values. (b) Mean duration of social and nonsocial behaviors for same genotype pairs is shown. All data are presented ± SEM. ** p ≤ 0.01

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
5.
Figure 7

Figure 7. Juvenile male MALTT mice can detect and differentiate nonsocial and social odors. From: Multiple autism-like behaviors in a novel transgenic mouse model.

A habituation/ dishabituation olfactory test was used to assess sense of smell. WT n = 13; TG n = 11. (a) Nonsocial odor 1 (banana or almond) was presented repeatedly followed by a second nonsocial odor (banana or almond) and mice were assessed for dishabituation on first presentation of novel odor and habituation by the third presentation of the odor. (b) Same task as (a) but nonsocial odors were replaced with social odors. For both tasks odor 1 and odor 2 were alternated pseudorandomly between subjects. All data are presented ± SEM. + denotes p≤0.05 for habituation to odor for both genotypes, # denotes p≤0.05 for detection of novel odor for both genotypes, open circle denotes Tg, closed circle denotes WT, ** p ≤ 0.01 for a between-subjects effect

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
6.
Figure 4

Figure 4. Juvenile MALTT mice show decreased social interest. From: Multiple autism-like behaviors in a novel transgenic mouse model.

Three-chamber partition task: (a) Diagram of the testing apparatus with two outer side chambers, each housing a novel object or stranger mouse behind perforated Plexiglas, and the center chamber where the subject is started. Two doorways allowed the subject to move freely between all chambers. (b) A MALTT or WT subject was allowed to explore the apparatus. Mean total duration each subject spent per chamber (including time at partitions within side chambers) is shown. choose to spend time investigating a stranger mouse or novel object through separate partitions, WT n = 8; TG n = 9. (c) Mean total duration at stranger or object partition is indicated. (d) Frequency of subject entry into each side chamber from the center chamber is shown. All data are presented ± SEM. * p ≤ 0.05

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
7.
Figure 5

Figure 5. Juvenile MALTT mice show social and social communication-related deficits. From: Multiple autism-like behaviors in a novel transgenic mouse model.

Direct social interaction task one: Two stranger mice were allowed to directly interact for 10 minutes while USVs were concomitantly recorded. (a) Mean duration of active, passive, and nonsocial behaviors for the subject mouse within a pair is shown. The subject of the pair was a WT in a WT + FVB/NJ stranger dyad, n = 8 pairs, or a TG in a TG + FVB/NJ stranger dyad, n = 9 pairs; (a–d). (b) Mean total USVs emitted by pairs is shown. (c) Representative event plot indicating (foreground) a pair’s USV events as they overlapped the subject mouse’s behavioral state (background). (d) Shown is the mean number of USVs emitted from the pair during the WT or TG subjects’ respective behavioral states. All data are presented ± SEM. * p ≤ 0.05, ** p ≤ 0.01

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
8.
Figure 8

Figure 8. MALLT mice show aberrant secondary ASD-associated behaviors. From: Multiple autism-like behaviors in a novel transgenic mouse model.

(a) A tactile sensitivity assay indicates minimum threshold for hind paw plantar surface detection of force for male mice, WT n = 8; TG n = 7. (b) Mean percent prepulse inhibition (PPI) response for male mice is shown. Data is collapsed over ages 17, 22, 28, 45 days. The same subjects tested for OFA were tested second on PPI. (c) Mean percent PPI response for female mice is shown collapsed over ages 17, 22, 28, 45 days. (d) Percent of mice exhibiting seizure activity in response to an audiogenic stimulus is shown for males WT n = 11, TG n = 19; females WT n = 9, TG n = 13. All data are presented ± SEM. * p ≤ 0.05, ** p ≤ 0.01

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.
9.
Figure 1

Figure 1. Initial characterization of the MALTT transgenic line. From: Multiple autism-like behaviors in a novel transgenic mouse model.

(a) A fluorescently labeled (FITC) anti-digoxigenin antibody indicated the location of the TyBS minigene insertion (left) and Geimsa staining indicated (black arrow) the labeled chromosome was X (right). (b) A schematic diagram of the mouse X chromosome indicates at minimum two copies of 4.4kb minigene construct (0.4kb MuLV sequence+1.8kb promoter region+2.2kb tyrosinase (tyr) cDNA) inserted in the non-coding region between genes Gm732 and Brwd3, as well as the 5’ integration sequence (black lettering: X chromosome, gray lettering:minigene). A Southern blot probe against the minigene is indicated in red. (c) A Southern hybridization image indicates 2 minigene positive bands (0.8kb, ~2.9kb) apparent in MALTT (lane 1) but not WT (lane 2) BanI-digested gDNA (left). (d) A single band (minimum 20kb) was apparent in MALTT (lane 1) but not WT (lane 2) BbsI-digested gDNA (right). (e) Exemplary sagittal and coronal Nissl-stained images of WT and MALTT brain sections are shown.

Shannon M. Hamilton, et al. Behav Brain Res. ;218(1):29-41.

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