1. Energy is expended by the wall cells of the ascending limb to pump Na+ ions from the filtrate to the interstitial fluid.
Therefore the interstitial fluid is hypertonic to the filtrate, but water stays inside the filtrate as the ascending limb is impermealbe to water.
300 mOsm + 200 mOsm = 500 mOsm
2. Fluid moves down the water-permeable descending limb, so water moves out by osmosis into the interstitial fluid.
The interstitial fluid is now hypotonic to the filtrate. Eventually so much water is drawn out of the descending limb that the interstitial fluid and the filtrate are isotonic, at 500 mOsm.
3. Protein pumps in the ascending limb pump out even more Na+ ions and raise the concentration of the interstitial fluid to 700 mOsm.
Filtrate in the descending limb reaches 700 mOsm.
This continues until a maximum of 1,200 mOsm is reached in the interstitial fluid.
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