URINE FORMATION AND

MEASUREMENT
 LO
Defined ginjal function
Defined urine formation
Defined how to make dilute and concentrate
urine
Defined renal clearance
 KIDNEY FUNCTIONS
Regulation of blood ionic composition

Regulation of blood pH

Regulation of blood volume

Regulation of blood pressure

Maintenance of blood osmolarity

Production of hormones

Regulation of blood glucose level

Excretion of wastes and foreign substances

 URINE FORMATION

Glomerular Tubular
Tubular secretion
filtration reabsorption

Tortora et al, 2009

Glomerular filtration
The filtration
membrane
Fenestration
(pore) of
glomerular
endothelial cell
Basal lamina of
glomerulus
Slit membrane
between pedicels
Glomerular Basal Lamina Podocyte
endothelial cell

Pore : 0,07- Contain Filtration slit

0,1 m collagen and Permit
Prevent proteoglicans passage
filtraton of Prevent molecule <
RBC & filtration of 0,006-0,007
platelet larger protein m (example,
Mesangial cell albmin)
contractile
cell help
glomerular
filtration
 Net Filtration Pressure

Tortora et al, 2009

 GLOMERULAR FILTRATION RATE
The amount of filtrate formed in all the renal
corpuscles of both kidneys each minute
+ 125 mL/min in males and 105 mL/min in females.
Homeostasis of body fluids requires that the kidneys
maintain a relatively constant GFR.
If the GFR is too high, needed substances may pass so
quickly through the renal tubules that some are not
reabsorbed and are lost in the urine.
If the GFR is too low, nearly all the filtrate may be
reabsorbed and certain waste products may not be
adequately excreted.
 The basic mechanisms that
regulate GFR
Neural
Regulation
Renal Hormonal
Autoregulation Regulation
(1) adjusting blood
flow into and out of
the glomerulus (2)
altering the
glomerular capillary
surface area for
filtration.
 Renal Regulation of GFR

Neural Hormonal
Renal regulation Regulation Regulation
Sympatic At rest Atrial natriuretic
Renal Angiotensin II
stimulation sympatheti reduces GFR
peptide (ANP)
Autoregulation Norepinephrine c stimulation increases GFR
is moderately
Afferent and
vasoconstriction low the efferent
The myogenic afferent and contriction Mesangial
mechanism : stretching by activation of cell relaxation
alfa1 receptors efferent
triggers contraction of increase
in the smooth arterioles are capilary
smooth muscle cells in dilated and
the walls of afferent muscle fibers of surface of
afferent renal filtration
arterioles autoregulatio
arterioles
n of GFR
Tubuloglomerular prevails
feedback
 TGF

Tortora et al, 2009

 TUBULAR REABSORPTION
Reabsorptionthe return of most of the
filtered water and many of the filtered solutes
to the bloodstream
About 99% of the filtered water is reabsorbed.
Epithelial cells all along the renal tubule and
duct carry out reabsorption, but proximal
convoluted tubule cells make the largest
contribution.
Solutes that are reabsorbed by both active and
passive processes include glucose, amino acids,
urea, and ions (Na, K, Ca2+, Cl, HCO3 and HPO4)
 TUBULAR SECRETION
The transfer of materials from the blood and
tubule cells into tubular fluid.
Secreted substances include ions (H, K, NH4
creatinine, and certain drugs such as
penicillin)
Tubular secretion has two important
outcomes:
(1) The secretion of H helps control blood pH.
(2) The secretion of other substances helps
eliminate them from the body.
 REABSORPTION ROUTE
Two route :
Paraceluller :
moving between
tubule cells
Transeluller :
passing through a
tubule cell

Tortora et al, 2009

 Transport Mechanisms
Primary active transport the energy derived from
hydrolysis of ATP is used to pump a substance
across a membrane; the sodium potassium pump is
one such pump.
Secondary active transport the energy stored in an
ions electrochemical gradient, rather than hydrolysis
of ATP, drives another substance across a membrane.
- Symporters are membrane proteins that move two
or more substances in the same direction across a
membrane.
- Antiporters move two or more substances in
opposite directions across a membrane.
 Water Reabsorption
Obligatory water reabsorption
90%
Water reabsorbed with solutes in tubular fluid
Occurs in the proximal convoluted tubule and the
descending limb of the loop because these segments
of the nephron are always permeable to water.

Facultative water reabsorption

10%
capable of adapting to a need
reabsorption is regulated by antidiuretic hormone
and occurs mainly in the collecting ducts
 Reabsoption and Secretion in PCT
Most solute reabsorption in the proximal
convoluted tubule (PCT) involves Na.
Na transport occurs via symport and antiport
mechanisms in the proximal convoluted
tubule.
Normally filtered glucose, AA, lactid acid,
water-soluble vitamin are completely
reabsorbed in the first half of the proximal
convoluted tubule by several types of Na
symporters located in the apical membrane
Reabsorption of glucose by Na-glucose
symporters in cells of PCT

Tortora et al, 2009

 In another secondary active transport process,
the Na/H antiporters carry filtered Na down
its concentration gradient into a PCT cell as H
is moved from the cytosol into the lumen
causing Na to be reabsorbed into blood and H
to be secreted into tubular fluid.
PCT cells produce the H needed to keep the
antiporters running in the following way.
Tortora et al, 2009
Tortora et al, 2009
 Solute reabsortion
in PCT promotes
osmosis of water by
transleluler and
paraceluller routes
PCT and ascending
loop cell very
permeable to water
because Aquaporin-
1
Tortora et al, 2009
 REABSORPTION IN LOOP HENLE
Tubular composition quite different with
glomerular filtrate because glucose, AA and
other nutrients are no longer presents
Reabsorbs :
15% of filtered water
20-30% of filtered Na and K
35% of filtered Cl
10-20% of filtered HCO3
and a variable amount of filtered Ca and Mg
Descending limbs reabsorb 15% of water, but
ascending limbs is impermeable to water
REABSORPTION IN LOOP HENLE
(THICK ASCENDING)
Main effect of
Na+-K+-2Cl-
sympoters is
reabsoption of
Na and Cl

Tortora et al, 2009

 REABSORPTION IN THE EARLY DCT
Reabsorb
10-15% of filtered water
5% of filtered Na+
5% of filtered Cl-
Reabsorption Na and Cl by Na+ Cl- symporter in
the apical membran
Reabsorption Na and Cl by Sodium-potasium
pumps and Cl leakage channel in basolateral
membrans
Major site where PTH stimulates reabsorption of
Ca
REABSORPTION AND SECRETION IN THE LATE
DCT AND COLLECTING DUCT
There are two type cell :
principal cell : reabsorb Na+, secrete K+
intercalatus cell : reabsorb K+ and HCO3-,
secrete H+
The amount of water and solute reabsorption
and secretion vary depending on the bodys
needs
Tortora et al, 2009
RESUME
Despospolulus et al, 2003
HORMONAL REGULATION OF TUBULAR
REABSORPTION AND SECRETION

RAA system

PTH ADH

ANP
 Decreases the glomerular By contraction of afferent
filtration arteriol

Enhances reabsorption of By stimulating activity of

RAA
Na, Cl and water in PCT Na/H antiporters

Stimulate principal cell in

collecting ducts reabsorb
Stimulates adrenal cortex to
more Na and Cl and secrete
release aldosterone
K + excreting less water
increases blood volume
 By increasing water
permeability of
principal cell in DCT
and collecting duct
Increase facultative
water reabsorption
Stimulates insertion of
aquaporin-2 on apical
membran via
exocytosis

ADH Kidneys can produce

When ADH as little as 400-500 ml
concentration maximal of very concentrate
urine

ADH defect diabetes Excrete up to 20 L of

insipidus very dilute urine daily
 Inhibit Na
reabsorption and
water in PCT and
collicting duct
ANP

Supress secretion of
Aldosteron and ADH
 Stimulated cell in
Increases Ca 2+
early DCT to
in blood
reabsorb Ca
PTH
Inhibits HPO42- Promoting
reabsorption in phosphate
PCT excretion
 FORMATION OF DILUTE AND
CONCENTRATE URINE
Eventhough our fluid intake can highly
variable, the total volume of fluid in our body
normally remains stable.
Highly fluid intake large volume of dilute
urine
Low fluid intake or loss of fluid small
volume of concentrate urine
PRODUCTION OF DILUTE URINE

Tortora et al, 2009

 PRODUCTION OF CONCENTRATE
URINE
When water intake is low or water loss is high
(such as during heavy sweating), the kidneys
must conserve water while still eliminating
wastes and excess ions
Under the influence of ADH, the kidneys
produce a small volume of highly
concentrated urine.
Urine can be four times more concentrated
(up to 1200 mOsm/liter) than blood plasma or
glomerular filtrate (300 mOsm/liter).
 Two main factors contribute to building and
maintaining this osmotic gradient:
(1) differences in solute and water
permeability and reabsorption in different
sections of the long loops of Henle and the
collecting ducts,
(2) the countercurrent flow of fluid through
tube-shaped structures in the renal medulla
Two types of countercurrent mechanisms
exist in the kidneys: countercurrent
multiplication and countercurrent exchange
COUNTERCURRENT MECHANISM
Tortora et al, 2009
The Countercurrent Multiplier System
Extrusion of NaCl from
ascending limb makes
surrounding interstitial
fluid more concentrated
Concentration multiplied
due to descending limb
Passively permeable to H2O
Fluid concentration increases
As surrounding interstitial
fluid becomes more
concentration

Deepest region of medulla

1,400mOsm
Ascending Limb Loop of Henle

Thin segment in depths

of medulla and thick
segment toward cortex
Impermeable to H2O;
permeable to salt
Thick segment Actively
Transports NaCl out of
filtrate

Active Transport of salt

filtrate becomes dilute
(100 mOsm) by end of
Loop of Henle
Active Transport in Ascending Limb
Na+ actively transported across basolateral membrane by Na+/ K+
pump
Cl- passively follows Na+ down electrical gradient
K+ passively diffuses back into filtrate
Transport of Ions in Ascending Limb
In thick segment Na+ and K+ together with 2 Cl- enter tubule cells
Na+ then actively transported out into interstitial space, Cl- follows
passively
K+ diffuses back into filtrate; some also enters interstitial space
 Vasa Recta
For countercurrent multiplier system to be effective:
Most of the salt extruded from ascending limbs must remain in
the interstitial fluid of the medulla
Most of the water that leaves descending limbs must be
removed by the blood

This is accomplished by the vasa recta

Thin-walled capillaries parallel LH of juxtamedullary nephrons
Walls permeable to water because of aquaporins channels,
NaCl, and urea but not plasma proteins
Therefore colloid osmotic (oncotic) pressure in vasta recta is
higher than in surrounding tissue fluid
results in movement of H2O from interstitial fluid into ascending vasa
recta that can be removed from the renal medulla
 Countercurrent Exchange in Vasa Recta
Important component of
countercurrent multiplier
Permeable to salt, H2O (via
aquaporins), and urea
Recirculates salt, trapping
some in medulla interstitial
fluid (maintains hypertonicity)
Reabsorbs H2O coming out of
descending limb
Descending section has urea
transporters
Ascending section has
fenestrated capillaries
Tortora et al, 2009
 Effects of Urea
Urea contributes to
high osmolality in
medulla
Deep region of
collecting duct is
permeable to urea
& transports it
The Role of Urea in Urine Concentration

1. Urea diffuses out of

inner collecting duct (in
renal medulla) into
interstitial fluid
2. Urea then passes into
ascending limb
Recirculates in interstitial
fluid of renal medulla
Urea and NaCl in
interstitial fluid make it
very hypertonic, so

3. Water leaves the CD by

osmosis
 Osmolality of Different Regions of the Kidney
Countercurrent multiplier
system in LH and
countercurrent exchange
in vasa recta
Creates hypertonic renal
medulla

Under influence of ADH

CD becomes more
permeable to H2O
Thus more H2O is drawn
out by osmosis into
hypertonic renal medulla
and peritubular capillaries
Osmotic Gradient in the Renal Medulla

Figure 25.13
Urine Concentrating Mechanisms
 Summary Counter-Current Multiplier
Peritubular fluid 1 2 3 4
Tubular fluid Fluid
300 300 300 300 300 300 300 300 300 300 300 300
Cortex

Medulla 300 300 300 300 400 200 400 400 200 300 400 200

Na+ H2 O Na+
Cl Cl
300 400 200 400 400 200
300 300 300 300 400 200
No osmotic H2 O
K+ K+
gradient
(mOsm) 300 400 200 400 400 200
300 300 300 400 400 400

300 300 300 300 400 200 400 400 200 400 400 400

Fluid Active transport Water moves Iso-osmotic state in More fluid Active
enters of Na+, Cl, K+ ions out of descending descending limb; enters tubule, transport of
tubule into medullary limb by osmosis osmotic difference pushing fluid Na+, Cl, K+
interstitial fluid between descending through by ions into
increases and ascending limbs bulk flow medullary
osmolarity interstitial
fluid increases
osmolarity

5 6 7

300 300 200 300 300 200 300 300 100

300 300 100

300 350 150 350 350 150
H2 O
500 500 300
300 350 150 350 350 150 Osmotic
gradient
H2 O established 900 900 700
400 500 300 500 500 300 (mOsm)
H2 O 1200 1200 1100

1400
400 500 300 500 500 300
1400

Water moves Iso-osmotic state in descending More water System

out of descending limb; osmotic difference enters tubule is in steady
limb by osmosis between descending and and process state
ascending limbs continues
RENAL CLEARANCE
 Urinary Excretion of Solutes
Sometimes tricky to calculate.
Things like univalent electrolytes, glucose, aas are
freely filterable.
However, if solute binds to protein, for example, then
its not (e.g. Ca2+, PO42-, Mg2+, PAH).
For these, you must measure plasma binding and
correct for the non-filterable fraction of solute.
Solute must also not be synthesized or degraded.
Ammonium is synthesised, and glutamate/glutamine
is degraded, as are other organic acids.
There are also complex combinations of reabsorption
and secretion with K+, uric acid and urea.
Must be careful, otherwise you will get inaccurate
measurements.

Urinary
Filtered Reabsorption Secretion by
Excretion = - +
Load by Tubules Tubules
of Solute
Filtration, reabsoption, and excretion rates of substances by the kidneys

Filtered Reabsorbed Excreted Reabsorbed

(meq/24h) (meq/24h) (meq/24h) (%)

Glucose (g/day) 180 180 0 100

Bicarbonate (meq/day) 4,320 4,318 2 > 99.9

Sodium (meq/day) 25,560 25,410 150 99.4
Chloride (meq/day) 19,440 19,260 180 99.1
Water (l/day) 169 167.5 1.5 99.1
Urea (g/day) 48 24 24 50
Creatinine (g/day) 1.8 0 1.8 0
 Introduction to Renal Measurement
There are 2 main categories of tests for assessing
renal function.
1. Modern imaging techniques macroscopic views of
renal blood flow, filtration and excretory function.
2. Measurements of renal clearance of various
substances to evaluate the ability of the kidneys to
handle solutes and water.
 Glomerular filtration is the flow of fluid from the
glomerular capillaries into the Bowmans capsule.
The volume filtrate formed per unit time is called
glomerular filtration rate
The rate of glomerular filtration averages 135-
180L/day in a normal adult.
 What is Clearance?
The clearance of a substance is the volume of plasma from
which that substance is cleared by the kidneys per unit time.
Each substance in the plasma has its own distinct clearance
value but it is always in the form of volume of plasma per
time.
To measure GFR
Substance able to achieve constant plasma concentration
Freely filtered at the glomerulus
Not reabsorbed or secreted
Filtered substance (eg inulin)= excreted substance
GFR x Pin = Uin x V
Use creatinine clearance as estimate of GFR
Creatinine clearance=Ucr x V/Pcr
Substance X is filtered and secreted but not reabsorbed.
Substance Y is filtered and some of it is reabsorbed.
Substance Z is filtered and completely reabsorbed.
 Measurement of GFR
GFR also assessed using principles of clearance.
GFR = vol. of fluid filtered into Bowmans capsule per
unit time.
Same equation, GFR is Cx if X has certain required
properties (i.e. Cinulin).

Conc. of X in urine
Volume of urine
formed in given

time
GFR = Ux x V
Glomerular Conc. of X in
filtration rate Px systemic blood
plasma
 Solutes used to measure GFR
Required properties are:
1. Solute is freely-filtered (conc. in Bowmans space = that in
blood plasma).
2. Tubules do not absorb, secrete or metabolize X.
Thus, amount of X in urine per unit time = that
which glomerulus filters per unit time.
WHAT GOES IN = WHAT COMES OUT!
INULIN is such a substance that satisfies all of these
criteria and is commonly used to measure GFR.
 Qualities of agents to measure GFR
Inulin: (Polysaccharide from Dahalia plant)
Freely filterable at glomerulus
Does not bind to plasma proteins
Biologically inert
Non-toxic, neither synthesized nor metabolized in
kidney
Neither absorbed nor secreted
Does not alter renal function
Can be accurately quantified
Low concentrations are enough (10-20 mg/100 ml
plasma)
 Drawbacks of Inulin
Most reliable method of measuring GFR, not useful clinically.
Inulin must be administered by IV to get relatively constant
plasma levels.
Chemical analysis of inulin in plasma and urine is technically
demanding.
Use radiolabelled compounds instead like radioactive Vitamin
B or EDTA.
However, these may also bind to proteins and distort results
slightly.
Problems of IV infusion of GFR marker avoided by using an
endogenous substance with inulin-like properties
CREATININE.
Renal handling of inulin

Amount filtered = Amount excreted

Pin x GFR Uin x V
Inulin Clearance
 Renal Plasma Flow
PAH delivery to kidney=PAH excretion
ERPF x P PAH =U PAH x V

Note: PAH only 85-90% is removed by kidney on

single pass, thus it will underestimate renal
plasma flow (filtered and secreted)
 Measurement of Renal Blood Flow

Not all blood delivered to glomerulus is filtered into

glomerular capsule
20% is filtered; rest passes into efferent arteriole and back into
circulation
Substances that aren't filtered can still be cleared by active
transport (secretion) into tubules
 p-aminohippurate (PAH)
There are certain special cases where the kidneys
completely clear X from plasma during a SINGLE
PASSAGE through them.
In this case, renal clearance of X = arterial renal
plasma flow.
p-aminohippurate (PAH) is such a special solute.
Thus, PAH clearance is a good estimate of renal
plasma flow.
 Renal handling of PAH
PAH is an organic acid that is
not usually present in the
body, so must give by IV
infusion.
Note that there is none left
in the renal vein - all cleared
in first pass.
Drawback is that mustnt
give too much PAH,
otherwise we overwhelm
the PAH secretory system
and the data can be
misleading.
 Use of PAH Clearance to Estimate Renal Plasma Flow

Paraminohippuric acid (PAH) is freely filtered and secreted

and is almost completely cleared from the renal plasma

1. amount enter kidney =

RPF x PPAH

2. amount entered =~ amount excreted

3. ERPF x Ppah = UPAH x V
~ 10 % PAH
ERPF = UPAH x V remains
PPAH
ERPF = Clearance PAH
 Creatinine clearance
Creatinine is an end product of muscle
metabolism
Muscle mass is constant; creatinine is constant
Normal 0.7 1.5 mg/ dL in plasma
Can then be compared to creatinine in urine
over 24 hour period to determine clearance

74
 Creatinine clearance is an indirect measure of
GFR and renal blood flow
Creatinine is neither reabsorbed nor secreted,
just freely filtered.
Amount excreted = amount filtered
Useful to monitor changes in chronic renal
function
Increases with trauma with massive muscle
breakdown

75
TERIMA KASIH