Mice were housed at the University of Pittsburgh Department of Laboratory Animals. Experimental procedures were approved by the University of Pittsburgh Institutional Animal Care and Use Committee. Mice were propagated in the C57Bl/6J background. All experimental mice were the product of crosses between male and female Muc1+/− mice. Genotyping was performed as previously described (39 (link)). All Muc1+/+ (control) mice were littermate controls. Mice were fed Prolab Isopro RMH 3000, LabDiet chow (1.09% Ca2+, 0.24% Mg2+, 0.94% K+, and 0.23% Na+) and water purified by reverse osmosis. They were maintained on a 12 h/12 h light/dark cycle.
Urine collection was performed as follows: to ensure voiding and to prevent volume depletion, animals were first injected with 7.5% (volume/body weight) sterile NSS and then placed in a metabolic cage. Urine voided in the first 30 min was discarded. Urine was then collected over the next 3.5 h. Animals were sacrificed, and bladder urine was aspirated and combined with urine collected in metabolic cages.
At time of sacrifice, mice underwent nonsurvival surgery under isoflurane anesthesia to collect blood, kidney, and duodenum specimens.
To examine urinary response to PTH receptor activation, mice were injected with the stable PTH analog, TPT. Mice were given an injection of vehicle alone (5% volume/body weight NSS, i.p.), and urine was collected in metabolic cages as six 1-h fractions. Two days later, mice were again injected with 5% body weight NSS, this time with 150 μg/kg TPT. Urine was again collected in metabolic cages as six 1-h fractions to assess urinary excretion of Na, phosphorus, and Ca.
Urine collection was performed as follows: to ensure voiding and to prevent volume depletion, animals were first injected with 7.5% (volume/body weight) sterile NSS and then placed in a metabolic cage. Urine voided in the first 30 min was discarded. Urine was then collected over the next 3.5 h. Animals were sacrificed, and bladder urine was aspirated and combined with urine collected in metabolic cages.
At time of sacrifice, mice underwent nonsurvival surgery under isoflurane anesthesia to collect blood, kidney, and duodenum specimens.
To examine urinary response to PTH receptor activation, mice were injected with the stable PTH analog, TPT. Mice were given an injection of vehicle alone (5% volume/body weight NSS, i.p.), and urine was collected in metabolic cages as six 1-h fractions. Two days later, mice were again injected with 5% body weight NSS, this time with 150 μg/kg TPT. Urine was again collected in metabolic cages as six 1-h fractions to assess urinary excretion of Na, phosphorus, and Ca.
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