Diluted and concentrated urine

- The kidneys responsible for cleaning of plasma from different substances and excrete them as urine
➔ The urine volume and concentration isn’t constant all the time
➔ It is dependent on the body status and food, for example:
✓ You may excrete 1.5 – 2 L of urine with concentration of 50 – 300 mosmol
 If you get high amount of water
✓ You may excrete 0.5 L of urine with concentration of 1200 mosmol
 If you get low amount of water
So, how we do this?

❖ Reabsorption in the different parts of nephron:

- Proximal convoluted tubule
➔ Reabsorb about 65 – 70 % of the substances
- Loop of Henle
➔ Thin descending
✓ Allow water reabsorption mainly
✓ Has low permeability to urea and ions
➔ Ascending part
✓ Thin segment
 Allow for Na reabsorption
 Impermeable for water
✓ Thick segment
 Allow for heavy Na reabsorption
 So, it is called the dilution segment
 Impermeable for water
 Notes:
- The Ascending part have many Na-K ATPase pump for active transportation of NaCl that have the
ability to generate 200 mosmol concentration difference
➔ Can pump Na from each point until the difference in the concentration between the inside & the
outside is 200 mosmol

- Distal convoluted tubule & Collecting ducts

➔ Moderate reabsorption of salts and Impermeable for water unless ADH is present
✓ If there is no ADH this part still impermeable to water.
✓ If there is ADH this part is highly permeable to water, especially in the cortex.
The loop of Henle is the part that is responsible for producing concentrated or diluted urine.

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 Diluted urine
- The nephron always produce a diluted urine
- How is it done?

- The amount of solutes in the filtrate is equal to that in the plasma so have the same osmolarity
(300mosmol) which is iso-osmotic
- In the PCT the reabsorption of both solutes & water is the same so it will still have the same
osmolarity (300mosmol)
- When it comes to the descending loop of Henle
➔ Water reabsorption is more than solutes reabsorption
➔ As we go downward the osmolarity become hypertonic until we reach tip with (1200mosmol)
How is the water reabsorbed?
▪ By osmosis the water diffuse from the area of high water to the area of low water
▪ The medullary-interstitial fluid has concentration gradient that increase as we go down in medulla
➔ From 300 at the beginning to 1200 at the end
✓ It maybe more than 1200.
Why as we go downward the osmolarity become hypertonic?
▪ By osmosis the water diffuse from the area of high water to the
area of low water
▪ Due to medullary-interstitial hypertonicity the water moves to the
outside, for example:
➔ 300 inside the loop and 400 outside
➔ The water in the inside will exit until it become 400 “equilibrium”

- In the Ascending loop of Henle

➔ Many Na-K ATPase pump for active transportation of NaCl
➔ As we go upward the osmolarity become hypotonic
(200mosmol)
Why as we go upward the osmolarity become hypotonic?
▪ By active transport the Na-K ATPase pump will pump Na to the outside, but it is impermeable for
water. Thus, the tonicity will decrease.
▪ At the border between the DCT & Ascending loop of Henle the filtrate will be hypotonic

- In the DCT and collecting duct

➔ There will be moderate NaCl reabsorption that decrease the osmolarity to reach (100mosmol)
➔ In the medullary collecting duct:
✓ Urea will be reabsorbed this will decrease the osmolarity to reach (50mosmol)
- Nephron produce diluted urine in cases of high water in the body
- So, it work by continuous reabsorb the solutes while failing to reabsorb water in distal tubule and
collecting duct, due to:
➔ Reducing ADH secretion leading to excrete urine with 50 mOsm/L.
➔ If large excess of water in the body, kidneys excrete up to 20 L/day
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 Concentrated urine
We produce a highly concentrated urine in conditions in which we want to preserve our fluid like
dehydration.
- How is it done?
1. Via the Countercurrent mechanism that involves 2 processes:
➔ Countercurrent Multiplication.
✓ To create a hypertonic medulla (medullary-interstitial
concentration gradient)
➔ Countercurrent exchanging.
✓ Maintenance of this medullary-interstitial
concentration gradient
✓ Via vasa recta
2. Via ADH

❖ Countercurrent mechanism
Countercurrent Multiplication
-

① Firstly, we have isotonic solution inside the tube in comparison to the outside with 300mosmol
② Na-K ATPase pump work and creates a 200mosmol difference
➔ The outside became 400mosmol
✓ Which is hypertonic to the first part
③ By osmosis the water will move until we reach the equilibrium making:
➔ The tube have the same osmolarity with outside
④ New volume of fluid is arrived which is 300mosmol & the 400mosmol fluid is pushed up
⑤ Again, Na-K ATPase pump work and creates a 200mosmol difference
➔ The outside became 500 and 350
✓ Concentration gradient start to be formed
⑥ By osmosis the water will move until we reach the equilibrium making:
➔ The tube have the same osmolarity with outside
⑦ New volume of fluid is arrived which is 300mosmol, and the previous will be repeated

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Countercurrent exchanging
- Factors keeping hyperosmolarity:
➔ Medullary blood flow is low 1 to 2% of total renal blood flow
✓ Which lead to make it very slow
➔ Vasa recta structure: descending and ascending segment which is U-
shape that:
✓ Minimizes loss of solute from interstitial
✓ There is a bulk flow of fluid & solutes into blood in which there
is exchange of water & solutes because it is capillary.
How is it Minimizes loss of solute from interstitial?
The reverse structure of it allows for preserve the hypertonicity in the medulla as the following:
- At the descending limb of vasa recta
➔ Blood enters isotonic then due to the concentration gradient:
✓ H2O will get out of vasa recta
✓ NaCl will get into vasa recta
➔ As blood goes downward it will be hypertonic until we reach the tip (osmolarity = 1200mosmol)

- The Ascending limb of vasa recta

➔ As blood goes upward due to the concentration gradient:
✓ H2O will get back into of vasa recta
✓ NaCl will get back out of vasa recta
➔ Blood enters isotonic then become hypertonic and leaves isotonic.
What is the unique in this? This is normal due to osmosis?!!
- The unique is the structure, and how it conserve the water and solutes inside the medulla without
take them out of it into circulation.

 Note:
- There is very poor lymphatic supply to the kidney to prevent the loss of hyperosmolarity

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Concentrated urine is not produced until now, so ADH will work.

❖ The role of ADH:

- ADH is hormone that is synthesized in the hypothalamus then stored into & secreted by posterior
pituitary gland.
- Released in case of low water in the body to preserve the water by decrease the amount of urine.

- Assume that someone have low water in his body then the amount of ADH will (increase)
➔ After it increased it will affect the DCT & collecting duct and increase their permeability to water
How ADH “vasopressin” increase DCT & collecting duct permeability to water?
- When vasopressin binds to its receptor V2R in the basolateral
membrane this will activates cascade of events that will lead to:

➔ Express of AQP-2 in the luminal membrane

➔ Express of AQP-3 or AQP-4 in the basolateral membrane

- This expressed channels that will allow water reabsorption from DCT & collecting duct efficiently
➔ This will lead to decrease the amount of water in the filtrate
➔ Then increase its osmolarity, & decrease water volume in it thus preserve the water in the body
The pathway of producing a concentrated urine:
- The amount of solutes in the filtrate is equal to that in the plasma so have the same osmolarity
(300mosmol) which is isotonic
- In the PCT the reabsorption of both solutes & water is the same so it will still have the same
osmolarity (300mosmol)
- When it comes to the descending loop of Henle
➔ Water reabsorption is more than solutes reabsorption
➔ As we go downward the osmolarity become hypertonic until we reach tip with (1200mosmol)

- In the Ascending loop of Henle

➔ Many Na-K ATPase pump for active transportation of NaCl
➔ As we go upward the osmolarity become hypotonic (200mosmol)

- In the DCT and collecting duct

✓ Due to the presence of ADH they will become very permeable for water
Water reabsorption in the DCT & collecting duct
- The majority of water reabsorption is occurred in the cortex by the cortical nephrons
➔ To conserve the medullary hypertonicity

- Then in the medullary collecting duct and due to the concentration gradient, the water will be
reabsorbed almost completely keeping the wastes with very low amounts of water.
➔ That’s why in very hot conditions when you don’t drink water your urine will become very dark.
5
The Role of Urea in the medullary concentration gradient and urea cycle
Urea reabsorption in the medullary collecting duct
- When the ADH is found the urea transporters UT1
will be expressed that allows the urea to reabsorbed
easily from it
➔ Going into the medulla increasing the
concentration of it
➔ Also, contribute to the formation of the
concentration gradient

- Then some of it go to the loop of Henle

➔ That will move again into the tubular system towards the collecting ducts
➔ Making a concentration gradient for urea to allow its exit

- This will be repeated again and again → this what we call the Urea Cycling
What is the role of urea in the medulla?
- Urea creates 500mOsm/L of the medullary concentration gradient and the others 700mosmol are
from the NaCl & water reabsorption
- This is why we excrete only about 50% of urea and other 50% are reabsorbed.
 Notes:
▪ The medullary concentration gradient may reach (10,000) in some animals like: Australian hopping
mouse.

Renal clearance of substances

- As we said the main renal function is to clean the plasma from the toxic substances
- But how efficient is that? How can we measure the renal function?
➔ We need a numbers for that, to compare and evaluate the renal function effectively

- How efficient is the kidney do their function?

➔ Through knowing the ability of the kidney to clean the bad substances from plasma into urine
This can be measured by:
- Renal clearance of substance, which defined as:
➔ The volume of plasma cleared of a particular substance per minute.
➔ The volume of plasma that is completely cleared of that substance by the kidneys
➔ ‫يعني قدرة الكلية حتى تنقي مقدار من البالزما من مادة معينة‬
➔ ‫احنا بنبحث عن كمية البالزما الي نقتها الكلية من مادة معينة بشكل كامل‬
➔ It refers not to the amount of the substance removed but to the volume of plasma from which
that amount was removed.
➔ It expresses kidneys’ effectiveness in removing various substances from internal environment.

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 .‫لنفرض انه المربع الكبير هاظ هو البالزما و المكعبات جوا هي الماده الي بدي اتخلص منها‬
‫لو نقيت الماده كلها من داخل المربع الكبير في وحدة الزمن بتكون الكليرنس = حجم المربع الكبير‬
‫لو نقيت الماده كلها من واحد من المربعات الصغيره بتكون الكليرنس = حجم المربع الي نقيته‬

- The effectiveness depends on:

➔ The value of renal clearance & The substance that is cleared
For instance:
- Assume that we have renal clearance of substance (X) that equal to GFR. Then, this substance will
go to the urine.
➔ If (X) is toxic substance, then:
✓ This is very good for our life & the renal function is efficient
➔ If (X) is glucose or AA, then:
✓ This is very bad for our life & the renal function is impaired
Calculated as:
quantity mL
(urine concentration of the substance ( )× urine flow rate ( ))
- Clearance rate of a substance (mL/min) =
mL of urine
plasma concentration of the substance (quantity/mL plasma)
min

- The plasma clearance rate varies for different substances, depending on how the kidneys handle
each substance:
Plasma Clearance Rate for a Substance Filtered but Not Reabsorbed or Secreted
- Assume that a substance X is freely filterable but is not reabsorbed or
secreted.
- As 125 mL/min of plasma are filtered and subsequently reabsorbed
➔ The quantity of substance X originally contained within the 125 mL is
left behind in the tubules to be excreted.
- Thus, 125 mL of plasma are cleared of substance X each minute.
Thus, the plasma clearance rate of a substance filtered but not reabsorbed or
secreted always equals the GFR
- This is applied for:
➔ Inulin (exogenous substance from ‫)نبات اسمه خرشوف القدس‬
✓ Doesn’t used to determine GFR
➔ Creatinine (endogenous substance)
✓ Often used to determine GFR

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Plasma Clearance Rate for a Substance Filtered and Reabsorbed
- Assume that a substance Y is freely filterable & reabsorbed.
- As 125 mL/min of plasma are filtered and subsequently reabsorbed
➔ The quantity of substance Y will decrease within the 125 mL.

- Some or all of a reabsorbable substance that has been filtered is

returned to the plasma
➔ Less than the filtered volume of plasma is cleared of the substance,
The plasma clearance rate of a reabsorbable substance is always less
than the GFR
- Reabsorbable substance are divided into:
➔ Completely reabsorbed
✓ None of the substance is cleared from plasma
✓ Clearance rate = Zero
✓ This is applied for: Glucose and Amino acids and Vitamins
➔ Partially reabsorbed
✓ Some of the substance is cleared from plasma
✓ Clearance rate less than GFR
✓ This is applied for: Urea (50%).

Plasma Clearance Rate for a Substance Filtered and Secreted

- Assume that a substance Z is freely filterable & secreted.
- As 125 mL/min of plasma are filtered and subsequently reabsorbed
➔ The quantity of substance z will increase within the 125 mL.
Why?
- Only 20% of the plasma entering the kidneys is filtered & the
remaining 80% passes unfiltered into the peritubular capillaries.
➔ The only way to clear it of any substance before being returned
to the general circulation is by secretion.
Thus, the plasma clearance rate for a secreted substance is always
greater than the GFR
- This is applied for:
➔ H+ & K+ & para-aminohippuric acid (PAH)

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Para-aminohippuric acid (PAH)
- It is filtrated by 20% & secreted by 80% thus completely excreted out of the body.
- Which means that:
➔ The amount of plasma that is cleaned from PAH is equal to the amount of
plasma that reach the kidney through the renal artery.
Thus, the plasma clearance for PAH is equal to the rate of plasma flow through the
kidneys.

 Notes:
▪ Clearance rates for inulin (or creatinine) and PAH can be used to
determine the filtration fraction.
▪ Practically no single volume of plasma is completely cleared of
substance.
▪ If a substance is completely cleared from plasma, then:
➔ The amount that delivered to kidneys = the amount excreted
➔ The clearance of substance = to the total renal plasma flow
➔ Like PAH

▪ If a substance is filtrated without secretion or reabsorption, then:

➔ Clearance = GFR
➔ Amt. Filtered = Amt. Excreted
➔ Like inulin & creatinine

▪ Calculations:
➔ Amt. delivered to the kidney = renal plasma flow x Plasma concentration of substance
➔ Amt. excreted from the body = Urine concentration of the substance x urine flow rate

The End