Comprehensive behavioral analysis of the Cdkl5 knockout mice revealed significant enhancement in anxiety- and fear-related behaviors and impairment in both acquisition and long-term retention of spatial reference memory methods

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    "name": "Comprehensive behavioral analysis of the Cdkl5 knockout mice revealed significant enhancement in anxiety- and fear-related behaviors and impairment in both acquisition and long-term retention of spatial reference memory methods",
    "description": "Protocol for Comprehensive behavioral analysis of the Cdkl5 knockout mice revealed significant enhancement in anxiety- and fear-related behaviors and impairment in both acquisition and long-term retention of spatial reference memory methods",
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        "position": 1,
        "name": "Cdkl5 -/Y mice show significantly enhanced anxiety-like behaviors",
        "text": "We performed the open field test for total duration of 120 minutes. In this test, exploration of a brightly lit, novel and unprotected environment is assessed during the first 5 minutes, and general activities during habituation to novelty are scored across a longer time [ ]. In the whole period, there was no significant difference of total distance traveled between two genotypes (F 1,42 = 0.063, p = 0.8038, two-way repeated measures ANOVA, ); however, during the first 5 minutes, Cdkl5 -/Y mice showed a marked decrease of the traveled distance compared to control mice ( ). The vertical activity of Cdkl5 -/Y mice was significantly decreased compared to control mice (F 1,42 = 17.73, p = 0.0001, two-way repeated measures ANOVA, ). The center time of Cdkl5 -/Y mice in the whole period was not significantly different from that of control mice (F 1,42 = 1.932, p = 0.1719, two-way repeated m..."
      },
      {
        "@type": "HowToStep",
        "position": 2,
        "name": "Cdkl5 -/Y mice exhibit a mild alteration in the gait",
        "text": "Previous studies reported impaired motor coordination of Cdkl5 KO mice, using the rotarod and CatWalk tests [, ]. We also identified an altered gait of our Cdkl5 -/Y mice. The stride pattern of the front paws (% of brake duration: F 1,38 = 5.92, p = 0.0198, one-way ANOVA, ) and the angles of hind paws (F 1,38 = 7.451, p = 0.0096, one-way ANOVA, ) were significantly different, and the stance widths of both front and hind paws were significantly shorter (front paw: F 1,38 = 4.58, p = 0.0388, one-way ANOVA, hind paw: F 1,38 = 7.708, p = 0.0085, one-way ANOVA, ). These results suggest a mild coordination disturbance in our Cdkl5 KO mice. The one-day rotarod test showed no significant difference in the latency to fall between genotypes (F 1,42 = 1.36, p = 0.2502, one-way ANOVA, )."
      },
      {
        "@type": "HowToStep",
        "position": 3,
        "name": "Cdkl5 -/Y mice exhibit a mild alteration in the gait",
        "text": "(A) Gait analysis of front paws. Cdkl5 +/Y, n = 22; Cdkl5 -/Y, n = 19. (i) Percent of stride duration. Cdkl5 KO mice show a significantly lower percentage of brake duration. (ii) Percent of stance duration. There is no significant difference in the duration of brake or propel between two genotypes. (iii) Cdkl5 -/Y mice show a significantly shorter stance width. (iv) There is no significant difference in the stride length between two genotypes. (v) There is no significant difference in the step angle between two genotypes. (vi) There is no significant difference in the paw angle between two genotypes. (B) Gait analysis of hind paws. Cdkl5 +/Y, n = 22; Cdkl5 -/Y, n = 19. (i) There are no significant differences in the duration of swing, brake, or propel between two genotypes. (ii) There is no significant difference in the duration of brake or propel between two genotypes. (iii) Cdkl5 -/..."
      },
      {
        "@type": "HowToStep",
        "position": 4,
        "name": "Cdkl5 -/Y mice show significantly enhanced anxiety-like behaviors",
        "text": "(A-D) The light/dark transition test. (A) Cdkl5 -/Y mice show a significantly decreased distance traveled in the light chamber and an increased distance in the dark chamber. (B) Cdkl5 -/Y mice show a significantly decreased stay time in the light chamber. (C) Cdkl5 -/Y mice show a significantly decreased number of transitions between chambers. (D) Cdkl5 -/Y mice show a tendency to increase in the latency to light chamber, but without statistical significance. (E-H) The open field test. (E) (i) Total distance traveled (cm) in 120 minutes. Each bin represents a mean value in 5 minutes. There is no significant difference between two genotypes (p = 0.8038). (ii) Total distance traveled during the first 5 minutes. Each bin represents a mean value in one minute. Marked reduction in the distance traveled of Cdkl5 -/Y mice is observed. (F) The vertical activity. Cdkl5 -/Y mice show a signific..."
      },
      {
        "@type": "HowToStep",
        "position": 5,
        "name": "Social interaction of Cdkl5 -/Y mice is uniquely altered",
        "text": "In order to evaluate the social interaction of Cdkl5 -/Y mice, we conducted the social interaction test in a novel environment (one-chamber test) ( ), three-chamber test ( ), and 24-hour home cage monitoring ( ). In the one-chamber test, we measured the total duration of contacts ( ), mean duration per contact ( ), number of contacts ( ), total duration of active contacts ( ), and distance traveled ( ). Active contacts denote physical contacts of two mice that run into each other and continue the locomotion for longer than 2 cm. Whereas physical contacts that reflect the social interaction correspond to spontaneous touching, rubbing, or sniffing behaviors toward each other, active contacts can occur just upon chance encounters during locomotion. The duration of active contacts should be greatly influenced by the activity level of the mice, and can be viewed as a parameter to reflect t..."
      },
      {
        "@type": "HowToStep",
        "position": 6,
        "name": "Social interaction of Cdkl5 -/Y mice is uniquely altered",
        "text": "(A-E) The social interaction test in a novel environment (one-chamber test). Cdkl5 -/Y mice show a significant increase in the total duration of contacts (A), a significant increase in the mean duration per contact (B), a significant decrease in the number of contacts (C), a significant decrease in the total duration of active contacts (D), and a significant decrease in the distance traveled (E). Cdkl5 +/Y, n = 11 pairs; Cdkl5 -/Y, n = 11 pairs. Data indicate means ± SEM. (F-K) The three-chamber social interaction test. Cdkl5 -/Y mice show a significant decrease in the total distance traveled (F), and a tendency toward decrease in the average speed (G). (H) Time spent in each chamber (sec). Cdkl5 -/Y mice show a significant preference for the center chamber, whereas Cdkl5 +/Y mice spend significantly more time in other chambers. (I) Time spent around cage (sec). Cdkl5 +/Y mice sp..."
      },
      {
        "@type": "HowToStep",
        "position": 7,
        "name": "Social interaction of Cdkl5 -/Y mice is uniquely altered",
        "text": "Twenty-four hour monitoring in the home cage under familiar conditions was conducted for one week to analyze the interaction and circadian activities of Cdkl5 KO mice. Social interaction was measured by counting the number of \"particle\", which represents an image of each mouse (see ); two particles indicated that the mice were not in contact with each other, and one particle indicated contact between them. Cdkl5 -/Y mice spent significantly more time staying together compared to control mice during daytime (mean number of particles F 1,20 = 75.79, p < 0.0001, two-way repeated measures ANOVA, ), but not in the nighttime (mean number of particles F 1,20 = 1.444, p = 0.2436, two-way repeated measures ANOVA, ). The genotype difference during the whole period was significant (F 1,20 = 14.166, p = 0.0012, two-way repeated measures ANOVA, ). The activity level of Cdkl5 -/Y mice w..."
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      {
        "@type": "HowToStep",
        "position": 8,
        "name": "Three-chamber social interaction test",
        "text": "The three-chamber social interaction test was performed as previously described [ ]. The testing apparatus consisted of a rectangular, three-chambered box and a lid with an infrared video camera (O'hara & Co., Tokyo). Each chamber was 20 cm x 40 cm x 22 cm and the dividing walls were made of clear Plexiglas, with small square openings (5 x 3 cm) allowing access into each chamber. An unfamiliar C57BL/6 male (stranger 1), that had had no prior contact with the subject mouse, was placed in one of the side chambers. The location of stranger 1 in the left vs. right side chamber was systematically alternated between trials. The stranger mouse was enclosed in a small, round wire cage, which allowed nose contact between the bars, but prevented fighting. The cage was 11 cm in height, with a bottom diameter of 9 cm, vertical bars 0.5 cm, and horizontal bars spaced 1 cm apart. The subject..."
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      "@type": "ScholarlyArticle",
      "headline": "Comprehensive behavioral analysis of the Cdkl5 knockout mice revealed significant enhancement in anxiety- and fear-related behaviors and impairment in both acquisition and long-term retention of spatial reference memory",
      "datePublished": "2018",
      "author": [
        {
          "@type": "Person",
          "name": "Kosuke Okuda"
        },
        {
          "@type": "Person",
          "name": "Keizo Takao"
        },
        {
          "@type": "Person",
          "name": "Aya Watanabe"
        },
        {
          "@type": "Person",
          "name": "Tsuyoshi Miyakawa"
        },
        {
          "@type": "Person",
          "name": "Masashi Mizuguchi"
        },
        {
          "@type": "Person",
          "name": "Teruyuki Tanaka"
        }
      ],
      "identifier": "10.1371/journal.pone.0196587"
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