Human insulin gene insertion in mice. Effects on the sleep-wake cycle?
JEAN-LOUIS VALATX (1), PHILIPPE DOUHET (2) and DANIELLE BUCCHINI (3) J.
Sleep Res. (1999) 8, Suppl. 1, 65-68
Table of Contents

Inroduction

Materials and methods

Results

Discussion

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MATERIALS AND METHODS

Animals

In a first series of experiments, nine adult male transgenic mice expressing HIg in the median habenular nucleus (DELTA- 168 mice) and six age-and sex-matched control C57BL/6 J (B6) mice were used. In a second set of experiments, two groups of transgenic mice were used, containing the HIg with altered 5'-flanking sequences resulting in either expression of the transgene only in pancreatic P-islets (DELTA-258) (n = 4) or in its nonexpression (A58) (n = 5) (Itier et al. 1996).

Surgery and recordings

Under pentobarbital anaesthesia (80 mg/kg, i.p.), mice were implanted with five cortical and three muscular electrodes, then caged singly in a Plexiglas jar and housed in a light-and temperature-controlled room (light on, 07.00 h; light off, 19.00 h; 23 + PC) with free access to water and food. All animals were treated according to guidelines approved by the European Communities Council Directive of 24 November 1986 (86/609/EEC). After 10 days of habituation to experimental conditions, continuous recordings were performed for at least 5 days, then the mice were subjected to a 10-h sleep deprivation period (09:00-19:00), using the water tank technique (Kitahama and Valatx 1980), and their recovery monitored over a further 2 days.

Data analysis

All recordings were scored visually, in 30-s epochs, for wakefulness (W), SWS, and paradoxical sleep (PS) using standard criteria (Valatx 1971). The data were then stored in a computer for further analysis using software designed in our own laboratory. The results are expressed as the mean + SD. Statistical analyses for the day, night, or full 24-h periods were performed by multiple factor analysis of variance, followed by multiple range test analysis. For sleep rebound analysis (first 24 h), Student's t-test was used, the significance level being set at P < 0.05.

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