Determination of Vitamin D-dependent calcium absorption by 45Ca gavage in the rat

Abstract

Precise determination of Vitamin D-dependent intestinal calcium absorption in longitudinal studies is problematic. We have assessed Vitamin D-dependent intestinal calcium absorption by 45Ca gavage. Rats were gavaged with a 1mL solution containing 45Ca (CaCl2, 9.3MBq/mL) maintained at 37°C. Total Ca concentration of the gavage fluid was optimised by comparing the absorption curves for fluids made up to 0.025, 2.025, 4.025 and 40.025mmol/L with 40CaCl2. The effect of varying dietary Ca on fractional Ca absorption was determined in rats fed semi-synthetic diets containing either 0.05%, 0.2%, 0.4% or 1.0% Ca for 50 days. Serum 1,25 dihydroxyvitamin D (1,25D) was determined by radioimmunoassay. Total gavage Ca of 0.025mmol/L achieved the highest peak fractional absorption and was adopted for all future experiments. Fifty days after allocation to the diets both fractional Ca absorption and 1,25D were highest in rats fed 0.05% Ca and lowest in those fed 1.0% Ca (absorption, P<0.05 and 1,25D, P<0.05). There was a strong logarithmic relationship between 1,25D and fractional Ca absorption (R2 0.69, P<0.001). Weekly repetition of the procedure did not cause a fall in haematocrit over 7 weeks. Radiocalcium (45Ca) absorption by gavage provides a simple measure of Vitamin D-dependent Ca absorption for repetitive use in longitudinal studies.