MECHANISM OF URINE FORMATION

There are four steps involved in the process:

I. ULTRA FILTRATION:
1. Thin membrane (2) separates blood in the capillary and cavity of the Malpighian
capsule. The pores are like holes of a sieve.
2. The tufts of glomeruli increase the surface area for filtration (about one and a half
sq. meter / kidney in mammals).
3. The renal artery is a short and wide one that branches immediately after entering
the kidney. Hence blood enters the kidney at a high pressure.
4. The lumen of the efferent arteriole is smaller than the lumen of afferent arteriole.
This leads to increase in blood pressure in each glomerulus.
5. Nervous and hormonal factors calibrate the diameter of the afferent and efferent
arterioles.
6. The hydrostatic pressure (A) of blood at the afferent glomeruli is about 60 mm Hg
in contrast to 25 mm Hg in the capillaries elsewhere in the body. This is sufficient
to overcome the resisting forces (B) existing in the plasma. They are:
1) Hydrostatic force exerted by fluid in Bowman’s space (15 mmHg)
2) Osmotic force due to presence of proteins in plasma (27mg). Proteins are almost
absent in filtrate. So, zero osmotic force in the filtrate.

Therefore, A minus B: 60mmHg – 42mmHg = 18 mmHg (Net filtration pressure)

There are about 10,00,000 glomeruli per kidney in a human being. Therefore, the
two kidneys put together filter 120ml / minute or 170 liters per day.

As a result of ultra filtration 20% of the plasma enters the Bowman’s capsule and
passes through the renal tubule. Only blood cells and High mol. wt. proteins (>60,000)
are retained in the blood and the rest are allowed to move into the filtrate. Eg: Gelatin
(35,000) and Egg albumin (34,500) are filtered while Serum albumin (67,500) and
Serum globulin (103,000) are not filtered. Glomerular filtrate at the end of first stage
contains about 95% water, low mol. wt. proteins, glucose, urea and salts.

Filtrate = Blood plasma - High mol. wt proteins (>60,000).

II. SELECTIVE REABSORPTION:

Resorption is carried out in the tubular part of the nephron which is richly
supplied with blood vessels coming from the glomerular artery. This enables formation
of hyper-osmotic urine (The glomerular filtrate is relatively dilute).
Many useful substances like glucose, amino acids, salts, water, electrolytes, are
reabsorbed and waste substances like urea and uric acid are allowed to be passed out
for subsequent elimination.
Renal Threshold denotes plasma concentration above which a given substance
appears in the urine. For example, in man the renal threshold for glucose is 120 –
160mg/dl plasma. Any amount above this is excreted and appears in the urine.
 Plasma clearance is the amount of a substance that normally appears in the urine.
Since no glucose appears in urine normally, the plasma clearance of glucose is
‘O’ and renal threshold is ‘100’. Penicillin presents an exact reverse situation. Its renal
threshold is ‘O’ and renal clearance is ‘100’.
Glucose is a high threshold substance and Penicillin is a non-threshold substance.
Such substances like urea, uric acid, salts and organic molecules which are partly
excreted and are also considered as non-threshold substances.
Water absorption is a passive process. It goes on due to establishment of a
concentration gradient due to reabsorption of substances like glucose, aminoacids,
sodium etc.
As sodium is actively transported into the tubular cells, water also passively
diffuses into the tubular cells from the lumen of the tubule. About 75% of filtrate is
reabsorbed before it reaches the Loop of Henle.
Some substances are transported passively along an electrical gradient. As
sodium is transported into the interstitial fluid from the lumen, chloride ions follow
sodium passively. The inner surface of the distal tuble is negatively charged. So,
positive ions from lumen of the tubule enter into the peritubular fluid. This takes place
mostly in the proximal tubule and the descending limb of loop of Henle. The walls of
these regions are thicker.
Some substances are exchanged continuously between of the tubule and the
peritubular fluid. Sodium is exchanged for Hydrogen ions throughout the nephron, a
process which aids in the maintenance of PH and electrolyte balance. In the distal tubule
Na+, Cl – and HCo3- are taken into the tubule wall while K+, H+ and NH4+ are put into
the filtrate flowing in the lumen.
Sugars, amino acids, lipids as well as many mono- & divalent- ions are absorbed
by the cells either by diffusion or active transport back into the circulation.
Urine is much more concentrated as compared to the filtrate. In urecotelic animals,
resorption of water is so extreme that the excreta is semi-solid (Birds, and Reptiles).

III. TUBULAR SECRETION:

Just before the urine is collected in its final from some substances are added into
it by this step. Ions like potassium are excreted into the lumen of the distal table when
the animal is placed under conditions of potassium loading. In birds and reptiles, Uric
acid is excreted entirely by secretion. In animals inhibiting dry environment the
glomeruli are reduced in size and / or number. Therefore urine formation goes on
almost entirely by this step.
As urine formed, mucus cells and epithelial cells are added before it is conveyed
into the bladder as urine.

IV. METABOLISM OF THE TUBULE CELLS:

The forth process is also important with respect to some substances. Renal
tubule cells synthesize for example Ammonium ion, which is later added to the lumen
fluid and excreted. It is utilized in regulation of plasma H+ concentration. Secondly,
tubule cells also catabolize peptides and eliminate them from the body.
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COUNTER CURRENT MULTIPLIER HYPOTHESIS FOR THE FORMATION
OF HYPEROSMOTIC URINE:

Types of mammalian kidneys: 1. Superficial nephron

2. Juxtaglomerular nephron
3. Juxtamedullary nephron
Most mammals produce hyperosmotic urine. The term ‘counter current’ is used because
the movements of fluid in the descending and ascending limbs are in opposite directions.
A small osmotic difference exists between the descending and the ascending
limbs of Loop of Henle. Due to counter current flow, the effect of osmotic difference at a
given level is further amplified as fluid courses through in the two regions of the tubule.
The walls of the descending limb of the Loop of Henle are permeable to the
sodium ions. As a result, sodium ions enter the glomerular filtrate. It is because sodium ion
concentration is higher in the interstitial fluid. This in turn is because of active reabsorption
taking place in the descending tubule. This is key to counter current multiplier system. In
other words, ascending limb actively cotransports chloride and sodium ions into descending
limb; but, at the same time being impermeable to water, does not allow water to follow the
salt. As more sodium enters the descending limb, the filtrate becomes increasingly
concentrated. Concentration reaches maximum at the tip of the loop.
Because of higher osmotic concentration in the interstitial fluid, more water
moves out of the tubule (the osmotic concentration of interstitial fluid is higher than that of
the tubule at each successive level). At the bend the osmotic concentration of interstitial
fluid is still slightly higher compared to tubular concentration.
The bend of the Loop of Henle lies deep in the medulla. As the fluid passes from
the bend of the loop into the ascending tubule and rises, the filtrate becomes progressively
less in its osmotic concentration by the time it reaches the cortex – medulla junction. As it
enters the distal convoluted tubule it is isotonic or hypotonic with the interstitial fluid. This
happens because of two reasons (explained earlier as well):
a) The ascending tubule is relatively impermeable to water. So water doesn’t move out
of the tubule.
b) Chloride is actively moved out of the tubular filtrate into the interstitial fluid.
Sodium passively follows chloride. Therefore, there is net movement of sodium
chloride from the tubular filtrate causing a drop in osmotic concentration.

The filtrate moves into the distal convoluted tubule which is again permeable to
water. As a result, water leaves the tubule and enters the interstitial fluid of the cortex
leading to gradual concentration of the filtrate.
Finally, the filtrate enters the collecting tubule. As it passes down, it gets more
and more concentrated. The final adjustment to osmotic concentration (in a small way) and
volume of urine in a major way is done here. Action of ADH hormone controls these events.
An increase in ADH level increases the permeability to water in distal and collecting
tubules. So, more water moves out into the interstitial fluid making the urine concentrated.
A decrease in the level of ADH level has the opposite effect leading to production of dilute
urine.
THE ROLE OF VASA RECTA: To maintain the counter current multiplier mechanism
efficiently, maintaining a gradient of osmotic concentration between interstitial fluid and
tubular filtrate is a mandatory requirement. This is done by the net work of capillaries (vasa
recta) formed from the efferent arteriole as it emerges out of the Bowman’s capsule. It has
two portions – descending thick set and ascending thin set of capillaries.
As blood passes through the thick capillary system of vasa recta present around
the ascending tubule, water diffuses outwardly because of higher osmotic concentration of
the interstitial fluid. As a result, osmotic concentration of blood increases as it reaches the
tip in the inner medullar region.
The ascending thin set of capillaries occurring around descending tubule group
together and form larger venules. They in turn join to form arcuate veins at the cortex –
medulla junction. Several of these join the renal portal vein and move the blood away from
the kidney.
When blood courses towards through the thin ascending set of capillary system
present around descending tubule towards the cortex, water diffuses inwardly. Sodium
chloride diffuses outwardly decreasing the osmotic concentration of blood again due to
osmotic gradient.
Whatever excess Na+ and Cl- ions are present in the interstitial fluid is removed
from the region along with water. Removal of that much amount of excess sodium, chloride
and water from the medulla maintains osmotic gradient necessary for counter current
multiplier effect.
This is how urine is concentrated in order to save the much precious water from
being wasted especially among those animals living in arid / desert conditions. It is common
knowledge that during summer volume of urine produced is very limited, concentrated and
yellowish in color where as during winter it is copious, watery and colorless.

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ROLE OF KIDNEY IN REGULATION OF SODIUM, POTASSIUM AND WATER:

RENAL SODIUM REGULATION: Sodium is freely filtered from the glomerular capillaries
in the Bowman’s capsule and there after actively reabsorbed back in the proximal tubule
and the loop of Henle: but, it is never secreted.

Therefore, Sodium excretion = Soduim filtered – Sodium reabsorbed

Sodium is a major extracellular solute / ion. It is a key solute responsible for maintaining
the extracellular osmotic concentration within the tissues. In case there is a decrease in the
body sodium levels, it leads to similar lowering of its levels in the extracellular fluid. As a
result, reabsorption of water from the glomerular filtrate into extracellular fluid decreases
leading to a drop in the blood volume and consequently a fall in BP. A sequence of events
occurs that restore normalcy.
Suppose there is an accident or diarrhea it results in a sever loss of blood and along with it
body sodium. Whenever there is a decrease in total body sodium, its excretion is lowered
so as to correct the situation in two ways:
1) Reduction in Glomerular Filtration Rate (GFR) and; 2) Increased Sodium reabsorption.
Lowering of the arterial pressure is sensed by baroreceptors that occur in the heart (atria)
and carotid sinus. These convey the information to the brain which in turn through the
sympathetic neurons elicits a response in the form of vasoconstriction of the afferent
arterioles. This leads to decrease in the GFR resulting in lesser filtration of sodium in to the
glomerular filtrate. This how sympathetic nerves influence the GFR.
Macula densa located on the inside of the tubule senses the sodium and chloride
concentration in the filtrate flowing past it. Whenever there is a decrease in the levels of
these ions and the consequent drop in pressure, it promotes increased secretion of renin by
JG cells.
JG cells that a specialized group of cells of the afferent arterial wall function as Intra-renal
Baroreceptors. They sense the drop in the pressure and respond by: a) bringing about
vasoconstriction of the afferent arteriole; b) increasing the production of renin which
triggers a chain of events that increase sodium reabsorption.
Macula densa and JG cells together constitute the JG apparatus (JGA). Both of them bring
about vasoconstriction of the afferent arteriole leading to lowering of the GFR. However,
these are not significant and efficient in increasing sodium reabsorption and restoration of
body sodium.

RENIN-ANGIOTENSIN-ALDOSTERONE SYSTEM (RAAS):

Renin Angiotensin-I Converting Enzyme Binding to receptors acts on

Angiotensinogen → Angiotensin-I → Angiotensin-II → Aldosterone
(From Liver) Tubule cells in Kidney Endothelial cells in Lung Cortical cells in Adrenal gland

Aldosterone acts on Kidney Tubules leads to ↑ Sodium Reabsorption & ↑Blood Vol & ↑BP

The action of aldosterone is on the distal tubule and cortical portion of the collecting duct.

NOTE: Cells of cardiac atria produce a peptide hormone that is antagonistic to the action of
aldosterone called Atrial Natriuretic Factor (ANF). It acts on kidney tubules and inhibits
Sodium reabsorption and also increases GFR resulting in increased Sodium excretion.

RENAL POTASSIUM REGULATION: Potassium is the most abundant intracellular ion. Only
2% of total body Potassium is extracellular. However, it is an extremely important one,
especially for excitability of nerve and muscle cells. An increase in extracellular potassium
depolarizes plasma membrane causing production in action potentials (nerve impulses) for
that moment. Following that, the membrane cannot produce further action potentials –
Leads to arrhythmic heart beat. On the other hand, a decrease hyperpolarizes plasma
membranes lowering their ability to generate action potentials – Leads to weakness in the
skeletal muscles abnormal conduction in the cardiac muscles.

Like Sodium, Potassium also is freely filtered from the glomerular capillaries in the Bowman’s
capsule and there after actively reabsorbed back in the proximal tubule and the loop of
Henle. However, distal tubule and the cortical portion of the collecting duct can actively
secrete Potassium to adjust the overall body levels during potassium overloading (eating
Potassium rich foods) under the direct influence of aldosterone. Na-K ATPase pump is
involved the regulation.
The cells of the adrenal cortex are very sensitive to Potassium ions. When there is an
increase in the level of these ions, the cortical cells increase production of aldosterone. The
hormone acts on the tubule producing: Potassium excretion and concurrent Sodium
reabsorption. The two cannot actions are interlinked and cannot be separated. On balance,
excess extracellular Potassium is more harmful compared to higher levels of Sodium.
Actually, in such circumstances, more insulin is released which enables the cells to take in
more of potassium ions thereby normalizing the extracellular Potassium in the body!!!

RENAL WATER REGULATION: As in the case of Sodium regulation, the baroreceptor

controlled reflexes are responsible for determining the volume of water excreted. However,
the major factor is not the rate of golmerular filtration (GFR), but the rate of water
reabsorbtion. This is under the control of Antidiuretic Hormone (ADH) also called as
Vassopressin released from neurohypophysis (Posterior Pituitary).
The pituitary releases ADH on the basis of the inputs received from baroreceptors and
osmoreceptors. They influence the pituitary via the hypothalamus located in the brain. A
decrease in the volume of extracellular fluid triggers RAAS which leads to increased
aldosterone production. In addition, they cause increased secretion of ADH. This increases
permeability of the walls of the collecting duct to water leading to increased water
reabsorption and decreased water excretion. If large volume of water is ingested, it would
lead to increased extracellular volume. In such a situation ADH secretion decreases leading
to increased water reabsoprtion.
The condition of increased urine excretion due to low levels of ADH is called as ‘Water
diuresis’. When ability to produce ADH is lost (usually due to damage to hypothalamus) it
results in a disease called ‘diabetes insipidus’ (literally meaning tasteless). This is distinct
from ‘diabetes mellitus’ caused due to insufficient production of insulin.

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