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Grip Strength (week 9) (Mice were fed on Mouse Breeder Diet (5021, Labdiet) from weaning): Neuromuscular function and muscle strength are assessed with a Bioseb grip strength meter. Three trials each of fore paws and all paws are performed immediately following each other.

Rotarod (week 10) (Mice were fed on Mouse Breeder Diet (5021, Labdiet) from weaning): Mice are assessed for motor activity and coordination by rotarod (Bioseb). Three 5 minute trials on a rod constantly accelerating from 4-40rpm are performed with a 15 minute inter-trial interval, and the latency and reason for ending each trial is recorded. The trial is ended when the mouse either falls off the rod, jumps off the rod, or performs a passive rotation (the mouse grasps the surface of the rod and rides around the rod instead of walking on it).

Indirect Calorimetry (week 12, male mice prioritised) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: Respiratory metabolic function is assessed using the LabMaster system (TSE-systems). Mice are housed individually in calorimetry cages for a period of approximately 21 hours including a 5 hour light phase acclimatisation period. This is followed consecutively by a 12 hour dark (19:30 - 07:30) and minimum 3 hour light phase. Cumulative food intake, activity (beam breaks), volume of oxygen consumed and volume of carbon dioxide produced are all measured. From this data the respiratory exchange ratio and energy expenditure are derived. Water bottles are weighed before and after the experiment to assess water consumption.

Hotplate (week 12) (Mice were fed on Mouse Breeder Diet (5021, Labdiet) from weaning): Mice are assessed for thermal pain perception using a hotplate (TSE systems). Each mouse is placed on the plate heated to 52?C, and the latency to the first response, and response type, are recorded. Mice that do not respond after 30 seconds are removed.

Open Field (week 9) (Mice were fed on Mouse Breeder Diet (5021, Labdiet) from weaning): Anxiety and exploratory drive are assessed over 20 minutes using open field apparatus (PhenoMaster ActiMot; TSE Systems)