Merging classical and modern genetic tools in the identification of disease genes

Abstract

We investigated if coloured microspheres could be used for repeated measurements of pancreatic islet blood flow in rats. An initial injection of 1.0-1.5 x 10⁵ microspheres (black colour), with a size of 10 or 15 µm, was made into the ascending aorta, while an arterial reference sample was collected from the femoral artery. Twelve min later, 1 ml of saline or 30% D-glucose was injected intravenously. Three min after this injection a second injection of 10- or 15-µm microspheres (green colour) was given. The animals were then killed, and the pancreas and adrenals were removed and samples (150-200 mg) were secured from the duodenum, ileum, colon, right kidney and liver. The microsphere contents were determined with the aid of a freeze-thawing technique and blood flow values were calculated. Our results suggest that 10-µm microspheres, but not 15-µm microspheres, provide reproducible islet and total pancreatic blood flow measurements when repeatedly injected. Values for the blood flow to the intestines, kidney and liver were less sensitive to the size of the microspheres. We conclude that repeated administration of 15-µm microspheres induces a high risk for erroneous islet and total pancreatic blood flow measurements, whereas two such measurements can be performed if 10-µm microspheres are used.