Explants of horses' hooves remained intact for up to 8 days when incubated in Dulbecco's modified Eagle medium (D-MEM) containing 25 mmol/l glucose but separated within 36 h when incubated in saline. The separation occurred between the basal epidermal cells and their basement membrane which is characteristic of the hoof separation that occurs in laminitis. Separation of hoof explants was prevented by addition of glucose to saline and was induced by adding 2-deoxyglucose or aminophenylmercuric acetate to D-MEM. Glucose consumption by the hoof explants was inhibited by 2-deoxyglucose and aminophenylmercuric acetate. The explants consumed relatively large amounts of glucose during the first 2 days of incubation and then little over the next 6 days. Despite the reduced glucose consumption, the hoof explants did not separate over 8 days of incubation. The results indicated that the integrity of the hoof explants was initially dependent on consumption of glucose and provide a possible explanation for the development of laminitis caused by conditions such as carbohydrate overload, acute inflammatory conditions, corticosteroid therapy and hyperlipidaemia. It would be expected that these conditions would induce a major hormonally-mediated metabolic shift away from glucose consumption by many peripheral tissues. It is suggested, therefore, that if the metabolic change occurred faster than the hoof tissue could adapt to an alternative energy substrate, then hoof separation and laminitis would occur.