Dr J.

Mohan

Renal Physiology

April, 2011

J. Mohan, PhD.
Lecturer,
Physiology Unit,
Faculty of Medical Sciences,
U.W.I., St Augustine.

Office : Room 105, Physiology Unit.

References:

Koeppen B.E. & Stanton B.A. (2010). Berne & Levy Physiology.
6th Edition. Mosby, Elsevier.

Marieb, E. & Hoehn, K. (2010). Human Anatomy & Physiology.

8th Edition, Pearson, Benjamin Cummings.

Stanfield, C.L. & Germann W.J. (2008). Principles of Human

Physiology. 3rd Edition, Pearson, Benjamin Cummings.

Hall, J.E. (2011). Guyton and Hall Textbook of Medical

Physiology. 12th Edition, Elsevier, Saunders.

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Dr J. Mohan

Physiology Objectives

1. Give an account of the main cellular mechanisms involved in

the modification of the glomerular filtrate as it flows through
the tubular segments of the nephron (transport and tubular
re-absorption of ions, organic solutes and water).

2. Summarise the events related to re-absorption of sodium

ions and water throughout the nephron.

Today’s Topics
• Solute and water transport along the nephron.

– Proximal Tubule
• Na+ Reabsorption.
• Water Reabsorption.
• Protein Reabsorption.
• Secretion of Organic Anions & Cations.

– Henle’s Loop

– Distal Tubule & Collecting Duct

• Regulation of NaCl & H20 Reabsorption

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Mechanisms of Urine Formation

• Urine formation
and adjustment of
blood composition
involves three
major processes

– Glomerular
filtration

– Tubular
reabsorption

– Secretion

Figure 25.10; Marieb & Hoehn, 2010

Solute and Water Transport along the Nephron

• Filtered at the glomeruli : ~ 180 L/day of essentially

protein-free fluid

• BUT < 1% of the filtered H20 & NaCl, and variable

amounts of other solutes are excreted in urine

Table 33.1, Koeppen & Stanton, 2010

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Solute and Water Transport along the

Nephron

• reabsorption & secretion = important processes by which

renal tubules modulate the volume & composition of urine

precise control of volume, osmolality & pH of the ECF & ICF

• mediated by transport proteins in cell membranes of the

nephron

• genetic & acquired defects in transport proteins
kidney

diseases + many transport proteins
important drug targets

Proximal Tubule
• reabsorbs :

– approximately 67% of filtered water, Na+, Cl-, K+, and

other solutes

– virtually all the glucose & amino acids filtered by the

glomerulus

• reabsorption of every substance, including water, is linked to

the operation of Na+,K+-ATPase in the basolateral membrane
of PT

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Na+ Reabsorption in 1st half of PT

Figure 33.1, Koeppen & Stanton, 2010

Na+ Reabsorption in 1st half of PT

• Ist half of PT

• Na+ entry to PT cell coupled to exit of H+ and entry of

HCO3-

– Na-H+ anti-port
• active transport of Na+ at basolateral membrane via
Na+,K+-ATPase
Na+ enters cell & H+ leaves at
apical membrane

– HCO3- transporters
• CO2 + H2O ↔ H2CO3 ↔ HCO3- + H+
• HCO3- leaves the cell at basolateral membrane

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Na+ Reabsorption in 1st half of PT

• Ist half of PT
• Na+ entry to PT cell coupled to entry of organic solutes
– Na+ enters proximal cells via several symporter
mechanisms, including Na+-glucose, Na+-amino acid,
Na+-Pi, & Na+-lactate
– glucose and other organic solutes that enter the cell with
Na+ leave the cell across the basolateral membrane via
passive transport mechanisms
– Na+-glucose symporter
• active tranport of Na+ at basolateral membrane via Na+/K+-
ATPase

• Na+ enters PT cell at apical membrane with glucose
• glucose leave the cell at basolateral membrane

Na+ Reabsorption in 1st half of PT

Summary

• reabsorption of Na+ in the first half of the PT is coupled to

that of HCO3- and a number of organic molecules

• reabsorption of Na+ - HCO3 & Na+ - organic solutes across

the PT
transtubular osmotic gradient
driving force for the
passive reabsorption of water by osmosis

• more water than Cl- is reabsorbed in the first half of the PT,
the [Cl-] in tubular fluid rises along the length of the PT

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Na+ Reabsorption in 2nd half of PT

Figure 33.3, Koeppen & Stanton, 2010

Na+ Reabsorption in 2nd half of PT

2nd half PT

• Na+ is mainly reabsorbed with Cl- across transcellular and

paracellular pathways

• Transcellular
– Na+ enters the cell across the apical membrane primarily
via the parallel operation of an Na+-H+ antiporter and one
or more Cl - - anion antiporters
– because the secreted H+ and anion combine in the
tubular fluid and reenter the cell, operation of the Na+-H+
and Cl- -anion anti-porters is equivalent to uptake of NaCl
from tubular fluid into the cell
–Na+ leaves the cell via Na+,K+-ATPase, and Cl- leaves
the cell and enters the blood via a K+-Cl- symporter in the
basolateral membrane

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Na+ Reabsorption in 2nd half of PT

• Paracellular

• paracellular NaCl reabsorption occurs because the rise in [Cl-]

in tubule fluid in the first half of the proximal tubule creates a
[Cl-] gradient (140 mEq/L in the tubule lumen and 105 mEq/L
in the interstitium)

• concentration gradient favors diffusion of Cl- from the tubular

lumen across the tight junctions into the lateral intercellular
space

• movement of negatively charged Cl- results in the tubular fluid

becoming positively charged relative to blood
positive
transepithelial voltage causes the diffusion of positively
charged Na+ out of the tubular fluid across the tight junction
into blood

Na+ Reabsorption in 2nd half of PT

• in the 2nd half of the proximal tubule, some Na+ and Cl- are
reabsorbed across the tight junctions via passive diffusion

• reabsorption of NaCl establishes a transtubular osmotic

gradient that provides the driving force for the passive
reabsorption of water by osmosis

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Na+ Reabsorption in PT
Summary

• reabsorption of Na+ by different mechanisms in the 1st & 2nd halves

of the PT

• in the 1st half of the PT reabsorption of Na+ is coupled to that of

HCO3- and a number of organic molecules (transcellular)

• in the 2nd half of PT , Na+ is mainly reabsorbed with Cl- across

transcellular and paracellular pathways

• approximately 67% of the NaCl filtered each day is reabsorbed in

the PT

• of this, two thirds moves across the transcellular pathway, whereas

the remaining third moves across the paracellular pathway

Water Reabsorption
• PT reabsorbs 67% of filtered water

Table 33.5, Koeppen & Stanton, 2010

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Water Reabsorption

Figure 33.4, Koeppen & Stanton, 2010

Water Reabsorption
• driving force for H20 reabsorption
– transtubular osmotic gradient established by reabsorption
of solute (e.g., NaCl, Na+-glucose)
– reabsorption of Na+ along with organic solutes, HCO3-&
Cl- from tubular fluid
lateral intercellular spaces

osmolality of the tubular fluid and
 the osmolality of
the lateral intercellular space

• PT- highly permeable to water
water reabsorbed via

osmosis
– apical & basolateral membranes of PT cells express
aquaporin water channels
water primarily reabsorbed
across PT cells
– some water also reabsorbed across tight junctions

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Water Reabsorption

• accumulation of fluid and solutes within the lateral intercellular

space
 hydrostatic pressure in this compartment
forces
fluid and solutes into the capillaries

• some solutes, especially K+ & Ca 2+ are dissolved in the

reabsorbed fluid, and so are reabsorbed by the process of
solvent drag (across tight junction)

• reabsorption of virtually all organic solutes, Cl- and other ions,

and water is coupled to Na+ reabsorption

• therefore, changes in Na+ reabsorption influence the

reabsorption of water and other solutes by the proximal tubule

Protein Reabsorption
• peptide hormones, small proteins & small amounts of large
proteins e.g. albumin are filtered by the glomerulus

• only a small percentage of proteins cross the glomerulus and

enter Bowman's space (i.e., the concentration of proteins in
the glomerular ultrafiltrate is only 40 mg/L)

• BUT, the amount of protein filtered per day is significant

because GFR is so high:

• Filtered Protein = GFR X [Protein] in ultrafiltrate

= 180L/day X 40 mg/L
= 7200 mg/day or 7.2 g/day

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Protein Reabsorption
• proteins undergo endocytosis either intact or after being
partially degraded by enzymes on the surface of proximal
tubule cells

• inside the cell, enzymes digest proteins
amino acids

leave the cell across the basolateral membrane by transport
proteins and are returned to the blood

• normally, this mechanism reabsorbs virtually all the proteins

filtered, and hence the urine is essentially protein free

• however, because the mechanism is easily saturated, an

increase in filtered proteins ( as could result from disruption of
the glomerular filtration barrier) causes proteinuria
(appearance of protein in urine)

Secretion of Organic Anions and Organic

Cations
• PT cells secrete organic cations & organic anions (end
products of metabolism that circulate in plasma)

• important in limiting the body's exposure to toxic compounds

derived from endogenous and exogenous sources

Tables 33.6 & 33-7, Koeppen & Stanton, 2010

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Secretion of Organic Anions and Organic

Cations
• exogenous organic compounds e.g. drugs & toxic chemicals

• many of these organic compounds can be bound to plasma

proteins and are not readily filtered
not eliminated from the
body via excretion after filtration alone

• secreted from the peritubular capillary
tubular fluid

• substances removed from plasma by both filtration &

secretion

Mechanisms of organic anion (OA-)

transport across the PT

Figure 33.5, Koeppen & Stanton, 2010

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Mechanisms of organic anion (OA-)

transport across the PT
• Basolateral membrane

– Na+/K+ATPase

– α-KG inside the cells via metabolism of glutamate and by an Na+-α-KG

symporter (i.e., a Na+-dicarboxylate transporter [NaDC])

– OA- -α-KG antiporter mechanisms : OAT1 & OAT3 (take up OA-s in

exchange for α-ketoglutarate (α-KG))

• resulting high intracellular concentration of OA- provides a driving force for

exit of OA- across the apical membrane membrane into tubular fluid

• Apical membrane

– OA-s are transported across the apical membrane by OAT4 & by MRP2
(multidrug resistance-associated protein 2)

Mechanisms of organic cation (OC+)

transport across the PT

Figure 33.6, Koeppen & Stanton, 2010

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Mechanisms of organic cation (OC+)

transport across the PT
• Basolateral membrane

– Na+/K+ATPase
– 4 pathways : passive diffusion & 3 uniporters (OCT1, OCT2, &
OCT3)

–
OC+s are transported into PT cells across the basolateral

membrane by three related transport proteins (OCT1, OCT2, and
OCT3) + diffusion
–
OC+s are taken up into the cell driven by cell negative
potential difference

• Apical membrane

– OC+s leave the cell across the apical membrane in exchange for
H+ by 2 OC+ - H+ antiporters (OCTN1, OCTN2 & MDR1)

Today’s Topics
• Solute and water transport along the nephron.

– Proximal Tubule
• Na+ Reabsorption.
• Water Reabsorption.
• Protein Reabsorption.
• Secretion of Organic Anions & Cations.

– Henle’s Loop

– DCT & Collecting Duct

• Regulation of NaCl & H20 Reabsorption

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Henle's Loop
• Henle's loop (LoH) reabsorbs approximately 25% of the
filtered NaCl and 15% of the filtered water (See Tables 33-4 &
33-5)

– thin ascending and thick ascending limbs of LoH

• reabsorption of NaCl
• impermeable to water

– descending thin limb

• water reabsorption via AQP1 water channels

• Ca2+ & HCO3- are also reabsorbed in the LoH

Henle's Loop
• thin ascending limb reabsorbs NaCl by a passive mechanism

• reabsorption of water, but not NaCl, in the descending thin

limb increases [NaCl] in the tubule fluid entering the thin
ascending limb

• as the NaCl-rich fluid moves toward the cortex, NaCl diffuses

out of the tubule fluid across the thin ascending limb into the
medullary interstitial fluid, down a concentration gradient
directed from the tubule fluid to the interstitium

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Mechanisms of solute transport across thick

ascending limb of LoH

Figure 33.7, Koeppen & Stanton, 2010

Mechanisms of solute transport across thick

ascending limb of LoH
• Basolateral membrane

– Na+/K+ATPase
– K+ - Cl- symporter
– HCO3- ?

• Apical membrane

– 1Na+-1K+-2Cl- symporter (NKCC2)

– K+ channel
– Na+-H+ antiporter

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Mechanisms of solute transport across

Henle's Loop

• increased NaCl transport by the thick ascending limb

increases the magnitude of the positive voltage in the lumen

• this voltage is an important driving force for the reabsorption

of several cations, including Na+, K+, Mg2+, & Ca2+, across
the paracellular pathway

Henle's Loop
Summary

• NaCl reabsorption across the thick ascending limb occurs via

the transcellular and paracellular pathways

• Fifty percent of NaCl reabsorption is transcellular and 50% is

paracellular

• Because the thick ascending limb does not reabsorb water,

reabsorption of NaCl and other solutes reduces the osmolality
of tubular fluid to less than 150 mOsm/kg H2O : "diluting
segment"

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Today’s Topics
• Solute and water transport along the nephron.

– Proximal Tubule
• Na+ Reabsorption.
• Water Reabsorption.
• Protein Reabsorption.
• Secretion of Organic Anions & Cations.

– Henle’s Loop

– DCT & Collecting Duct

• Regulation of NaCl & H20 Reabsorption

Distal Tubule and Collecting Duct

• The distal tubule and collecting duct :

– reabsorb approximately 8% of the filtered NaCl

– secrete variable amounts of K+ & H+

– reabsorb a variable amount of water (≈8% to 17%)

(See Tables 33-4 & 33-5)

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Mechanism for reabsorption of Na+ & Cl- in

the early segment of the DT

Figure 33.8, Koeppen & Stanton, 2010

Distal Tubule and Collecting Duct

• Early DT
– reabsorbs Na+, Cl- & Ca++
– impermeable to water

• Basolateral membrane
– Na+/K+ATPase
– Cl- channels

• Apical membrane
– Na+Cl- symporter

• dilution of tubular fluid begins in the thick ascending limb and

continues in the early segment of the DT

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Distal Tubule and Collecting Duct

• Late DT & CD
– principal cells
• reabsorb NaCl & water and secrete K+

– intercalated cells
• secrete either H+ or HCO3- ( acid-base balance)
• reabsorb K+

Mechanisms of transport in the

late DT & CD

Cl-

Figure 33.9, Koeppen & Stanton, 2010

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Mechanisms of transport in Principal Cells

Principal cells
• Basolateral membrane
– Na+,K+-ATPase
– K+ channels
– AQP 3&4

• Apical membrane
– epithelial Na+-selective channels (ENaCs)
– K+ channels
– AQP2

Mechanisms of transport in Principal Cells

• Cl- reabsorbtion
– reabsorption of Na+ generates a negative luminal voltage
across the late DT & CD, which provides the driving force
for reabsorption of Cl- across the paracellular pathway

• H20 reabsorbtion
– water reabsorption is mediated by the AQP2 water
channel located in the apical plasma membrane & by
AQP3 and AQP4 located in the basolateral membrane of
principal cells

– in the presence of antidiuretic hormone (ADH), water is

reabsorbed
– in the absence of ADH, the DT & CD reabsorb little water

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Mechanisms of transport in Principal Cells

• K+ secretion
– K+ is secreted from blood into tubular fluid by principal
cells in two steps :
– (1) uptake of K+ across the basolateral membrane is
mediated by the action of Na+,K+-ATPase
– (2) K+ leaves the cell via passive diffusion
• K+ channels
• K+ diffuses down its concentration gradient through apical cell
membrane K+ channels into tubular fluid
• although the negative potential inside the cells tends to retain
K+ within the cell, the electrochemical gradient across the
apical membrane favors secretion of K+ from the cell into
tubular fluid

Mechanisms of transport in the

late DT & CD

Cl-

Figure 33.9, Koeppen & Stanton, 2010

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Mechanisms of transport in Intercalated

Cells
• Intercalated cell

– Basolateral membrane
• HCO3- (passive diffusion)

– Apical membrane
• H+ leaves the cell
• H+-K+-ATPase mediates K+ reabsorbtion

Today’s Topics
• Solute and water transport along the nephron.

– Proximal Tubule
• Na+ Reabsorption.
• Water Reabsorption.
• Protein Reabsorption.
• Secretion of Organic Anions & Cations.

– Henle’s Loop

– DCT & Collecting Duct

• Regulation of NaCl & H20 Reabsorption

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Regulation of NaCl & Water Reabsorption

NaCl reabsorption

• hormones
– angiotensin II, aldosterone, NE, Epi, natriuretic peptides,
and uroguanylin, dopamine & adrenomedullin

• Starling forces

• Glomerulotubular balance

H20
• ADH (direct)

Regulation of NaCl & Water Reabsorption

NaCl reabsorption

Angiotensin II

• Major Stimulus
–  renin

• Site of Action
– PT, thick ALoH, DT/CD

• Effect on Transport
–  reabsorption of NaCl & H20

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Regulation of NaCl and Water Reabsorption

NaCl reabsorption

Aldosterone

• Major Stimulus
–  AgII,  plasma [K+]

• Site of Action
– thick ALoH, DT/CD

• Effect on Transport
–  reabsorption of NaCl & H20
–  secretion of K+

Mechanisms for Reabsorption of NaCl by

Aldosterone
• early distal tubule
–  # of the Na+-Cl- symporter

• principal cells – DT/CD

– (1)  the amount of Na+,K+-ATPase in the basolateral
membrane
– (2)  expression of the Na+ channel (ENaC) in the apical
cell membrane
– (3)  Sgk1 (serum glucocorticoid-stimulated kinase) levels

 the expression of ENaC in the apical cell membrane
– (4) stimulating CAP1 (channel-activating protease, also
called "prostatin"), a serine protease that directly activates
ENaCs by proteolysis

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Mechanisms for Reabsorption of NaCl by

Aldosterone
• reabsorption of Na+
negative luminal voltage across the
distal tubule and collecting duct.

• this negative voltage in the lumen provides the

electrochemical driving force for reabsorption of Cl- across the
tight junctions (i.e., paracellular pathway) in the distal tubule
and collecting duct

• through its stimulation of NaCl reabsorption in the collecting

duct, aldosterone also indirectly increases water reabsorption
by this nephron segment

Mechanisms for Reabsorption of NaCl by

Natriuretic peptides (ANP & BNP &
• Major Stimulus
urodilatin)
–  ECFV;  BP

• Site of Action
– Medullary CD

• Effects
–  reabsorption of NaCl & H20
 excretion
–  ADH-stimulated water reabsorption across the collecting
duct
–  secretion of ADH from the posterior pituitary

• actions of ANP & BNP mediated by the activation of membrane-

bound guanylyl cyclase receptors
 cGMP

• Urodilatin > profound natriuresis & diuresis than ANP >

profound natriuresis & diuresis than BNP

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Mechanisms for Reabsorption of NaCl by

NE & Epi
• Major Stimulus
–  ECFV;  BP

• Site of Action
– PT, thickALoH, DT/CD

• Effects
–  reabsorption of NaCl & H20

Mechanisms for Reabsorption of H20 by

ADH
• Major Stimulus
–  Posm;  ECFV

• Site of Action
– DT/CD

• Effects
–  reabsorption of H20

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Mechanisms for Reabsorption of NaCl &

H20 by Starling’s forces

Figure 33.9, Koeppen & Stanton, 2010

Mechanisms for Reabsorption of NaCl &

H20 by Starling forces
• Starling forces that favor
movement from the
interstitium into the
peritubular capillaries are
πpc & Pi

• the opposing Starling forces

are πi & Ppc

• major Starling force driving • Recall : glomerular filtration


reabsorbtion = π pc (high
oncotic pressure of the
peritubular capillary blood)
the [protein ] and π of glomerular
capillaries; this blood leaves
glomerular capillaries
efferent
arterioles
peritubular
capillaries

April 07 & 11, 2011 29

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Mechanisms for Reabsorption of NaCl &

H20 by Starling forces
• therefore, peritubular capillary oncotic pressure (πpc) is
partially determined by the rate of formation of the glomerular
ultrafiltrate

• E.g.  in GFR, at constant RPF in the afferent arteriole


[plasma proteins] in the efferent arteriole & peritubular
capillary
 π pc
• πpc is directly related to the filtration fraction :
FF = GFR
RPF

•  FF resulting from a  GFR, at constant RPF
 π pc

 net reabsorption of solute and water across the PT

G-T Balance
Glomerulotubular (G-T) balance

• major mechanism for regulating solute & water reabsorption

by the PT

• describes the balance between filtration (glomerulus) and

reabsorbtion (PT)

• depends on Starling forces

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G-T Balance
• spontaneous changes in GFR markedly alter the filtered load
of Na+ (filtered load = GFR × [Na+] in the filtered fluid)

• e.g.  GFR
 filtered Na+
 urinary excretion of Na+

disturbed Na+ balance, if Na+ reabsorption is not rapidly
adjusted

• however, spontaneous changes in GFR do not alter Na+

excretion in urine or Na+ balance because of the
phenomenon of G-T balance

G-T Balance
• when body Na+ balance is normal (i.e., ECF volume is
normal), G-T balance refers to the fact that reabsorption of
Na+ & water increases in proportion to the increase in GFR
and filtered load of Na+

• therefore, a constant fraction of the filtered Na+ and water is

reabsorbed from the PT despite variations in GFR

• the net result of G-T balance is to reduce the impact of

changes in GFR on the amount of Na+ & water excreted in
urine

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G-T Balance
• 2nd mechanism

– increase in the filtered load of glucose & amino acids

 reabsorption of Na+ & H20

– reabsorption of Na+ in the 1st half of the proximal tubule

is coupled to that of glucose & amino acids

–the rate of Na+ reabsorption therefore partially depends

on the filtered load of glucose & amino acids

– as the GFR and filtered load of glucose & amino acids

increase, reabsorption of Na+ & water also rises

G-T Balance
Summary of G-T balance

•  in GFR

–  FF
 π pc
 solute & H20 reabsorbtion in PT

–  filtered glucose & aa
 reabsorbtion Na+ in PT

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G-T Balance

Summary of GT-balance

•  in GFR

–  FF
 π pc
 reabsorbtion in PT
–  filtered glucose & aa
 reabsorbtion Na+ in PT

• Proportionality of filtration and reabsorbtion is maintained i.e.

there is glomerular tubular balance

• Na+ homeostasis (and ECF volume and blood pressure)

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