Age-related changes in behavior in C57BL/6J mice from young adulthood to middle age methods
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Decreased locomotor activity and altered anxiety-like behaviors in older C57BL/6J mice
The open field test is widely used to measure locomotor activity and anxiety-like behavior [ ]. In the first 5-min period of the test, there were significant effects of Age on the distance traveled (Fig.; F 3,1451 = 8.51, p < 0.0001) and stereotypic counts (Fig.; F 3,1451 = 8.049, p < 0.0001), which achieved study-wide significance. Subjects in older age groups traveled significantly shorter distances than those in younger age groups (8-12 mo < 4-5 and 2-3 mo, p = 0.0009 and p < 0.0001, respectively; 6-7 mo < 2-3 mo, p = 0.0012). The 6-7- and 4-5-month-old subjects showed significantly more stereotypic counts than 2-3-month-old subjects ( p = 0.0007 and p < 0.0001, respectively). Durin...
Decreased social contacts in older C57BL/6J mice
The social interaction test, in which two unfamiliar mice are placed together in a novel chamber, is used to assess social behavior. There were significant effects of Age on the distance traveled (Fig.; F 3,546 = 8.48, p < 0.0001), number of contacts (Fig.; F 3,546 = 11.71, p < 0.0001), and total duration of active contacts (Fig.; F 3,546 = 8.58, p < 0.0001), all of which reached study-wide significance. Older age subjects traveled significantly shorter distances than subjects in the youngest age group (8-12, 6-7, and 4-5 mo < 2-3 mo, p < 0.0001, p = 0.0043, and p = 0.0055, respectively). The number of contacts in the older age groups was significantly lower than those in the younger age groups (8-12 and 6&#...
Decreased immobility in older C57BL/6J mice
The Porsolt forced swim test, in which a mouse is placed into a cylinder filled with water for 10 min a day for 2 days, is performed to measure immobility time as an index of depression-related behavior. One-way ANOVAs revealed significant effects of Age on immobility time (Fig.: day 1, F 3,1084 = 3.18, p = 0.0234; day 2, F 3,1084 = 5.64, p = 0.0008) and distance traveled (Fig.: day 1, F 3,1084 = 5.51, p = 0.0009; day 2, F 3,1084 = 1.06, p = 0.3671). On day 1, immobility times in older age groups were lower than those in the youngest age group, although these differences did not survive a Bonferroni correction (8-12, 6-7, and 4-5 mo < 2-3 mo, p = 0.0255, p = 0.03, and p = 0.0383, respe...
General health and neurological screen
The general health check and neurological screen was conducted as previously described [ - ]. Body weight and rectal temperature were measured. Neuromuscular strength was assessed using the grip strength and wire hang tests. A grip strength meter (O'Hara & Co., Tokyo, Japan) was used to assess forelimb grip strength. Mice were lifted and held by their tail so that their forepaws could grasp a wire grid. The mice were then gently pulled backward by the tail until they released the grid. The peak force applied by the forelimbs of the mouse was recorded in Newtons (N). Each mouse was tested three times, and the largest value was used for statistical analysis. In the wire hang test, the mouse was placed on a wire mesh that was then inverted, and the latency to fall from the wire was recorded with a 60 s cut-off time.
Social interaction test in a novel environment
Social interaction test [, ] was conducted as previously described [ ]. Two mice of same genotype that were previously housed in different cages, were placed into a box together (40 × 40 × 30 cm; O'Hara & Co., Tokyo, Japan) and were allowed to explore freely for 10 min. Mouse behavior was analyzed automatically using ImageSI software (see Section, "Image analysis"). The total duration of contacts (s), number of contacts, total duration of active contacts (s), mean duration per contact, and total distance traveled (cm) were measured. The active contact was defined as follows: images were captured at three frames per second, and distance traveled between two successive frames was calculated for each mouse. If the two mice contacted each other and the distance traveled by either mouse was 5 cm and more, the behavior was...
Startle response/prepulse inhibition test
A startle reflex measurement system (O'Hara & Co., Tokyo, Japan) was used to measure startle response to a loud noise and prepulse inhibition of the startle response. A test session began by placing a mouse in a plastic cylinder where it was left undisturbed for 10 min. White noise (40 ms) was used as the startle stimulus for all trial types. The startle response was recorded for 400 ms starting with the onset of the startle stimulus. The background noise level was 70 dB. The peak startle amplitude was used as a dependent variable. A test session consisted of six trial types (e.g., two types of startle-stimulus-only trials, and four types for prepulse inhibition trials). The intensity of the startle stimulus was either 110 or 120 dB. The prepulse sound was presented 100 ms before the onset of the startle stimulus, and its intensity was 74 or 78...
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The open field test is widely used to measure locomotor activity and anxiety-like behavior [ ]. In the first 5-min period of the test, there were significant effects of Age on the distance traveled (Fig.; F 3,1451 = 8.51, p < 0.0001) and stereotypic counts (Fig.; F 3,1451 = 8.049, p < 0.0001), which achieved study-wide significance. Subjects in older age groups traveled significantly shorter distances than those in younger age groups (8-12 mo < 4-5 and 2-3 mo, p = 0.0009 and p < 0.0001, respectively; 6-7 mo < 2-3 mo, p = 0.0012). The 6-7- and 4-5-month-old subjects showed significantly more stereotypic counts than 2-3-month-old subjects ( p = 0.0007 and p < 0.0001, respectively). During the first 5 min, there were trends toward significant effects of Age on the vertical activity (Fig.; F 3,1451 = 2.25, p = 0.0806) and center time (Fig.; F 3,1451 = 2.38, p = 0.0681). The older age groups tended to exhibit reduc...
The social interaction test, in which two unfamiliar mice are placed together in a novel chamber, is used to assess social behavior. There were significant effects of Age on the distance traveled (Fig.; F 3,546 = 8.48, p < 0.0001), number of contacts (Fig.; F 3,546 = 11.71, p < 0.0001), and total duration of active contacts (Fig.; F 3,546 = 8.58, p < 0.0001), all of which reached study-wide significance. Older age subjects traveled significantly shorter distances than subjects in the youngest age group (8-12, 6-7, and 4-5 mo < 2-3 mo, p < 0.0001, p = 0.0043, and p = 0.0055, respectively). The number of contacts in the older age groups was significantly lower than those in the younger age groups (8-12 and 6-7 mo < 2-3 mo, p = 0.0002 and p < 0.0001, respectively; 8-12 and 6-7 mo < 4-5 mo, p = 0.0079 and p = 0.0017, respectively). Subjects in older age groups showed a significantly shorter duration of...
The Porsolt forced swim test, in which a mouse is placed into a cylinder filled with water for 10 min a day for 2 days, is performed to measure immobility time as an index of depression-related behavior. One-way ANOVAs revealed significant effects of Age on immobility time (Fig.: day 1, F 3,1084 = 3.18, p = 0.0234; day 2, F 3,1084 = 5.64, p = 0.0008) and distance traveled (Fig.: day 1, F 3,1084 = 5.51, p = 0.0009; day 2, F 3,1084 = 1.06, p = 0.3671). On day 1, immobility times in older age groups were lower than those in the youngest age group, although these differences did not survive a Bonferroni correction (8-12, 6-7, and 4-5 mo < 2-3 mo, p = 0.0255, p = 0.03, and p = 0.0383, respectively). Similar results were obtained for immobility time on day 2 (8-12 and 4-5 mo < 2-3 mo, p = 0.0013 and p = 0.0013, respectively). On day 1, the 6-7-month-old subjects swam longer distances than the 4-5- and 2-3-month-o...
The general health check and neurological screen was conducted as previously described [ - ]. Body weight and rectal temperature were measured. Neuromuscular strength was assessed using the grip strength and wire hang tests. A grip strength meter (O'Hara & Co., Tokyo, Japan) was used to assess forelimb grip strength. Mice were lifted and held by their tail so that their forepaws could grasp a wire grid. The mice were then gently pulled backward by the tail until they released the grid. The peak force applied by the forelimbs of the mouse was recorded in Newtons (N). Each mouse was tested three times, and the largest value was used for statistical analysis. In the wire hang test, the mouse was placed on a wire mesh that was then inverted, and the latency to fall from the wire was recorded with a 60 s cut-off time.
Social interaction test [, ] was conducted as previously described [ ]. Two mice of same genotype that were previously housed in different cages, were placed into a box together (40 × 40 × 30 cm; O'Hara & Co., Tokyo, Japan) and were allowed to explore freely for 10 min. Mouse behavior was analyzed automatically using ImageSI software (see Section, "Image analysis"). The total duration of contacts (s), number of contacts, total duration of active contacts (s), mean duration per contact, and total distance traveled (cm) were measured. The active contact was defined as follows: images were captured at three frames per second, and distance traveled between two successive frames was calculated for each mouse. If the two mice contacted each other and the distance traveled by either mouse was 5 cm and more, the behavior was considered an "active contact."
A startle reflex measurement system (O'Hara & Co., Tokyo, Japan) was used to measure startle response to a loud noise and prepulse inhibition of the startle response. A test session began by placing a mouse in a plastic cylinder where it was left undisturbed for 10 min. White noise (40 ms) was used as the startle stimulus for all trial types. The startle response was recorded for 400 ms starting with the onset of the startle stimulus. The background noise level was 70 dB. The peak startle amplitude was used as a dependent variable. A test session consisted of six trial types (e.g., two types of startle-stimulus-only trials, and four types for prepulse inhibition trials). The intensity of the startle stimulus was either 110 or 120 dB. The prepulse sound was presented 100 ms before the onset of the startle stimulus, and its intensity was 74 or 78 dB (20 ms). Four combinations of prepulse and startle stimuli were used (74-110, 78-110, 74-120, and 78-120 dB). Six blocks of the six trial types were presented in a pseudorandom order such that each trial type was presented once within a block. The average inter-...
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