MECHANISMS OF

PASIVE AND ACTIVE

TRANSPORT
THROUGH THE
MEMBRANES
 OSMOSIS
 Osmosis is the diffusion of water through a
semi-permeable membrane.

 More specifically, it is the movement of water across a

semi-permeable membrane from an area of high
water potential (low solute concentration) to an area of low
water potential (high solute concentration).

 It is a physical process in which a solvent moves, without

input of energy, across a semi-permeable membrane
(permeable to the solvent, but not the solute) separating
two solutions of different concentrations.

 Osmosis releases
energy, and can be
made to do work (as
when a growing tree root
splits a stone).

Diffusion and Osmosis are

both types of PASSIVE
TRANSPORT - that is,
no energy is required for
the molecules to move
into or out of the cell.
 https://www.youtube.com/watch?v=GQnp
W82TXhA

 http://www.youtube.com/watch?v=rR7NO
SRyzhM
 http://www.youtube.com/watch?v=l7Piud
E0QbM&list=TLQQhP6FqpP3oxIieROW
ERnBEguHz1WM4Y
Net movement of solvent is from the less-concentrated (hypotonic) to
the more-concentrated (hypertonic) solution, which tends to reduce the
difference in concentrations. This effect can be countered by increasing
the pressure of the hypertonic solution, with respect to the hypotonic.
Partially-Permeable Membranes

• A partially-permeable membrane will allow

certain molecules to pass through it, but not
others.
Partially permeable
• Generally, small membrane
particles can pass
through…

…but large
particles
cannot 6
 Partially-permeable
More free water molecules membrane
on this side of membrane

Water-solute
particle is too large
to pass through
membrane

Free water molecules diffuse in this direction

7
Osmosis

• Osmosis is the diffusion of free water

molecules…
•… from a region of high concentration of
free water molecules…
• … to a region of low concentration of free
water molecules…
• …across a partially-permeable
membrane…
• …until they are evenly spread out.

8
Basic principle of an osmometer

The
osmotic pressure is
defined to be the
pressure required to
maintain an
equilibrium, with no
net movement of
solvent.

The solvent will diffuse from the compartment with smaller

concentration to that with higher concentration until the
concentration will be equal. The selective membrane doesn’t the
allow substance dissolved in the solvent (glucose, etc) to pass
the membrane.
 Osmotic flux is evidenced by the liquid level in the right
part of the “U” tube.
 Hydrostatic pressure in the tube is a measure of the
osmotic pressure.

 Osmosis may be opposed by increasing the pressure in

the region of high solute concentration with respect to
that in the low solute concentration region.
 The force per unit area, or pressure, required to prevent
the passage of water through a selectively-permeable
membrane and into a solution of greater concentration
is equivalent to the osmotic pressure of the solution,
or turgor.
 Osmotic pressure is a colligative property, meaning that
the property depends on the concentration of the solute
but not on its identity.
Osmotic pressure - the laws of
osmosis by Van’t Hoff and Pfeffer.

 π = ρ g h, where ρ = density of the solution, g= 9,8 m/s2,

h = the high level of the liquid in the right tube.

First law. At a constant temperature, the osmotic

pressure of a diluted solution is proportionally with
its molecular concentration.

C
 K
K= constant
C= concentration in percent

M M= molecular weight
  Second law. At the temperature lower then
40ºC the osmotic pressure vanes
proportionally with the absolute
temparature.
π = K’ C T
c n
  K K CT  K T or V  nRT
M V
V= volum of solution, n = nr. of mols of dissolved
substance, R= universal gas constant= 8310 J/KmolK.
Van’t Hoff formula
Osmotic Pressure

Laws of Osmotic Pressure :

1. The Osmotic Pressure is directly
proportional to the concentration of
the solute.
2. The Osmotic Pressure is directly
proportional to the absolute
temperature.
The osmole
 The osmotic pressure doesn’t depend on the nature of
the dissolved substance; it depends only on the
concentration, temperature and molecule weight.
 The ability of the solutes to cause osmosis and osmotic
pressure is mesured in terms of osmoles. The osmole
is a measure of total number of particles.
 One gram-mole of non-diffusible and non-ionisable
substance is equal to 1 osmole.
 Osmolare solutions = solutions containing the same
number of osmotic active particles, equal with
NA = 6,023 x 1023 mol-1. (Avogadro number).
Osmolality / osmolarity
 The osmole is too large unit for satisfactory use in
biology and medicine. Therefore, the term of
miliosmole is commonly used:
 1 mOsm= 1/1000 Osm.
 The osmolal concentration of a solution is called
osmolality when the concentration is expressed in
osmoles/ Kg of water .
 It is called osmolarity when it is expressed as osmoles/
liter of solution.
 The osmotic law is modified by some factors of
correction in case of electrolytes –taking into account
also the dissociation degree, and in the case of
macromolecular solutions – taking into consideration
also the strength of the forces of interaction between
the molecules of solute and solvent.
 Osmosis can also be explained using the notion of entropy, from
statistical mechanics.
 As above, suppose a permeable membrane separates equal amounts
of pure solvent and a solution. Since a solution possesses more
entropy than pure solvent, the second law of thermodynamics states
that solvent molecules will flow into the solution until the entropy of the
combined system is maximized. Notice that, as this happens, the
solvent loses entropy while the solution gains entropy.
 Equilibrium, hence maximum entropy, is achieved when the entropy
gradient becomes zero, and dissolution takes place.
 Pure water is more ordered than water in a solution; thus, from an
entropic standpoint it takes some net energy to move water molecule
from a disordered solution and "pack it in" with pure water.
 This is the same explanation as to why the disordered air does not
spontaneously separate and order into oxygen and nitrogen, it would
take energy for this to happen.
 Additionally, particle size has no bearing on osmotic pressure, as this
is the fundamental postulate of colligative properties.
Examples of osmosis
 Osmotic pressure is the main cause of support in many plants. The
osmotic entry of water raises the turgor pressure exerted against the
cell wall, until it equals the osmotic pressure, creating a steady state.

 When a plant cell is placed in a hypertonic solution, the water in the cells
moves to an area higher in solute concentration and the cell shrinks, and in
doing so, becomes flaccid. (This means the cell has become plasmolysed -
the cell membrane has completely left the cell wall due to lack of water
pressure on it; the opposite of turgid.)

 Also, osmosis is responsible for the ability of plant roots to draw water from
the soil. Since there are many fine roots, they have a large surface area,
and water enters the roots by osmosis.

 Osmosis can also be seen when potato slices are added to a high
concentration of salt solution. The water from inside the potato moves to
the salt solution, causing the potato to shrink and to lose its 'turgor
pressure'. The more concentrated the salt solution, the bigger the
difference in size and weight of the potato slice.
 In unusual environments, osmosis can be very harmful to
organisms. For example, freshwater and saltwater aquarium fish
placed in water of a different salinity than that they are adapted to
will die quickly, and in the case of saltwater fish, dramatically.
 Another example of a harmful osmotic effect is the use of table
salt to kill leeches and slugs.

 Suppose an animal or a plant cell is placed in a solution of

sugar or salt in water:
 If the medium is hypotonic — a dilute solution, with a higher water
concentration than the cell — the cell will gain water through
osmosis.
 If the medium is isotonic — a solution with exactly the same water
concentration as the cell — there will be no net movement of
water across the cell membrane.
 If the medium is hypertonic — a concentrated solution, with a
lower water concentration than the cell — the cell will lose water
by osmosis.
Osmosis and Diffusion

 In the real world

In plants, osmosis occurs for example
at root hairs, allowing the uptake of water
from the soil.
Osmosis and Diffusion
 In the real world……….
 In humans, osmosis occurs in the kidneys to recover the water form waste
materials of the body. The kidneys regulate the concentration of water in the
blood plasma.

 Kidney dialysis
Effect of different
solutions on blood cells
Egg Osmosis

 http://www.youtube.com/watch?v=SSS3
EtKAzYc
 http://www.youtube.com/watch?v=7-QJ-
UUX0iY
Reverse osmosis

 Reverse osmosis is a separation process that

uses pressure to force a solvent through a
membrane that retains the solute on one side and
allows the pure solvent to pass to the other side.
 More formally, it is the process of forcing a
solvent from a region of high solute concentration
through a membrane to a region of low solute
concentration by applying a pressure in excess of
the osmotic pressure.
Application: Drinking
water purification
Around the world, household drinking water purification systems,
including a reverse osmosis step, are commonly used for
improving water for drinking and cooking.
Such systems typically include a number of steps:
 a sediment filter to trap particles including rust and calcium
carbonate
 optionally a second sediment filter with smaller pores
 an activated carbon filter to trap organic chemicals and chlorine,
which will attack and degrade TFC reverse osmosis membranes
 a reverse osmosis (RO) filter which is a
thin film composite membrane (TFM or TFC)
 optionally a second carbon filter to capture those chemicals not
removed by the RO membrane
 optionally an ultra-violet lamp for disinfecting any microbes that
may escape filtering by the reverse osmosis membrane
 Dialysis
 Reverse osmosis is similar
to the technique used in
dialysis, which is used by
people with kidney failure.
 The kidneys filter the blood,
removing waste products
(e.g. urea) and water, which
is then excreted as urine.
 A dialysis machine mimics
the function of the kidneys.
The blood passes from the
body via a catheter to the
dialysis machine, across a
filter.
Principle of dialysis
 Dialysis works on the principles of the diffusion of solutes and ultrafiltration of
fluid across a semi-permeable membrane.
 Blood flows by one side of a semi-permeable membrane, and a dialysate fluid
flows by the opposite side. Smaller solutes and fluid pass through the
membrane.
 The blood flows in one direction and the dialysate flows in the opposite. The
counter-current flow of the blood and dialysate maximizes the concentration
gradient of solutes between the blood and dialysate, which helps to remove
more urea and creatinine from the blood.
 The concentrations of solutes (for example potassium, phosphorus, and urea)
are undesirably high in the blood, but low or absent in the dialysis solution and
constant replacement of the dialysate ensures that the concentration of
undesired solutes is kept low on this side of the membrane.
 The dialysis solution has levels of minerals like potassium and calcium that are
similar to their natural concentration in healthy blood. For another solute,
bicarbonate, dialysis solution level is set at a slightly higher level than in normal
blood, to encourage diffusion of bicarbonate into the blood, to act as a pH
buffer to neutralise the metabolic acidosis that is often present in these
patients.
 The levels of the components of dialysate are typically prescribed by a
nephrologist according to the needs of the individual patient.
 Osmotic pressure of
blood plasma
 Blood plasma is an aqueous solution containing different ions (Na+,
K+, Ca2+…), small molecules molecule non-dissociated
(glucose,aminoacids) and proteins- macromolecules (albumin,
globulin). Each type of molecules contributes with his own osmotic
pressure, the sum representing the colloid-osmotic pressure (or
oncotic pressure)


 π = ∑ πmolec.nedisoc + ∑ πioni+ ∑ πplcol

Where ∑ πplcol = ∑ πmacromol = ∑ πalbumine + ∑ πglobuline

65% 35%
 Colloid osmotic pressure of blood
plasma is 22 Torr at la 37ºC
(1 Torr = 1mmHg)
 It represent that pressure developed
by the blood plasma in contact with
pure water through a selective
membrane.
 The lateral pressure that the blood exerts against the walls of the
blood vessels is called blood pressure.
 It is caused by
a) contraction of the ventricles

b) resistance to the passage of blood through arterioles and capillaries.

The pressure existing in the arteries is called arterial blood pressure.
The blood pressure is high during systole and is called systolic blood
pressure. It is low during diastole and it is called diastolic blood
pressure. The difference between these two pressures is called pulse
pressure

 The blood pressure is high in the aorta situated near the heart. It
gradually decreases as it goes away from the heart. It is low in the
capillairies.
 The blood pressure is minimum in the veins. For clinical purpose, the
blood pressure is measured from the large arteries of the arm. The
instrument used is called a sphygmomanometer.
 The systolic blood pressure in an ideal man is 120 mm of Hg and
the diastolic blood pressure is 80 mm of Hg. Blood pressure is
measured in millimeters of mercury (mm. Hg)
Here, filtration refers to leaving the
capillary and entering the interstitial fluid.
Absorption, on the other hand, is the
uptake of fluid or nutrients from the
interstitial fluid into the capillary
It’s also important to remember that
different pressures exist at the arterial end
vs. the venous end
Edema mechanism
 Edema- formerly known as dropsy or hydropsy - is an abnormal
accumulation of fluid beneath the skin, or in one or more cavities
of the body.
 Generally, the amount of interstitial fluid is determined by the
balance of fluid homeostasis, and increased secretion of fluid into
the interstitium or impaired removal of this fluid may cause edema.
Five factors can contribute to the formation of edema:
 It may be facilitated by increased hydrostatic pressure or,
 reduced oncotic pressure within blood vessels;
 by increased blood vessel wall permeability as in inflammation;
 by obstruction of fluid clearance via the lymphatic; or,
 by changes in the water retaining properties of the tissues
themselves. Raised hydrostatic pressure often reflects retention of
water and sodium by the kidney.[1]
 Generation of interstitial fluid is regulated by the forces of the
Starling equation.

 The Starling equation is an equation that illustrates the role of

hydrostatic and oncotic forces (the so-called Starling forces) in
the movement of fluid across capillary membranes.
 Capillary fluid movement may occur as a result of two processes:
 diffusion
 filtration

 Starling's equation only refers to fluid movement across the

capillary membrane that occurs as a result of filtration.
 In the glomerular capillaries, there is a net fluid filtration of 125
ml/min (about 180 litres/day).
 In the rest of the body's capillaries, there is a total net
transcapillary fluid movement of 20 ml/min (about 28.8 litres/day)
as a result of filtration. This is several orders of magnitude lower
than the total diffusional water flux at the capillary membrane, as
that is about 80,000 litres/day.
 The Starling equation was formulated in 1896 by the British
physiologist Ernest Starling.
Starling's equation

where:
([Pc − Pi] − σ[πc − πi]) is the net driving force,
Kf is the proportionality constant, and
Jv is the net fluid movement between compartments.

By convention, outward force is defined as positive, and inward force is defined

as negative. The solution to the equation is known as the net filtration or net fluid
movement (Jv). If positive, fluid will tend to leave the capillary (filtration). If
negative, fluid will tend to enter the capillary (absorption). This equation has a
number of important physiologic implications, especially when pathologic
processes grossly alter one or more of the variables.
According to Starling's equation, the movement of fluid
depends on six variables:

 Capillary hydrostatic pressure ( Pc )

 Interstitial hydrostatic pressure ( Pi )
 Capillary oncotic pressure ( πc )
 Interstitial oncotic pressure ( πi )
 Filtration coefficient ( Kf )
 Reflection coefficient ( σ )
 Filtration coefficient
 The filtration coefficient is the constant of proportionality. A high
value indicates a highly water permeable capillary. A low value
indicates a low capillary permeability.
 The filtration coefficient is the product of two components:
 capillary surface area
 capillary hydraulic conductance

 Reflection coefficient
 The reflection coefficient is often thought of as a correction
factor. The idea is that the difference in oncotic pressures
contributes to the net driving force because most capillaries in the
body are fairly impermeable to the large molecular weight
proteins. (The term ultrafiltration is usually used to refer to this
situation where the large molecules are retained by a
semipermeable membrane but water and low molecular weight
solutes can pass through the membrane).
 Many body capillaries do have a small permeability to proteins (such
as albumins). This small protein leakage has two important effects:
1. the interstitial fluid oncotic pressure is higher than it would otherwise
be in that tissue
2. not all of the protein present is effective in retaining water so the
effective capillary oncotic pressure is lower than the measured
capillary oncotic pressure.
 Both these effects decrease the contribution of the oncotic pressure
gradient to the net driving force. The reflection coefficient (σ) is used
to correct the magnitude of the measured gradient to 'correct for' for
the ineffectiveness of some of the oncotic pressure gradient. It can
have a value from 0 up to 1.
 Glomerular capillaries have a reflection coefficient close to 1 as
normally no protein crosses into the glomerular filtrate.
 In contrast, hepatic sinusoids have a low reflection coefficient as they
are quite permeable to protein. This is advantageous because
albumin is produced in hepatocytes and can relatively freely pass
from these cells into the blood in the sinusoids. The predominant
pathway for albumin and other proteins to enter the circulation is via
the lymph.
 Some albumin escapes from the capillaries and enters the
interstitial fluid where it would produce a flow of water
equivalent to that produced by a hydrostatic pressure of +3
mmHg. Thus, the difference in protein concentration would
produce a flow of fluid into the vessel at the venous end
equivalent to 28 - 3 = 25 mmHg of hydrostatic pressure. The
total oncotic pressure present at the venous end could be
considered as +25 mmHg.

 In the beginning (arteriolar end) of a capillary, there is a net

driving force (([Pc − Pi] − σ[πc − πi])) outwards from the
capillary of +9 mmHg. In the end (venular end), on the other
hand, there is a net driving force of -8 mmHg.

 Assumed that the net driving force declines linearily, then

there is a mean net driving force outwards from the capillary
as a whole, which also results in that more fluid exits a
capillary than re-enters it. The lymphatic system drains this
excess.
 http://www.youtube.com/watch?v=fh50nq
YU3i8
Glomerular filtration rate (GFR)
 is the volume of fluid filtered from the renal (kidney) glomerular
capillaries into the Bowman's capsule per unit time.
 Glomerular filtration rate (GFR) can be calculated by measuring
any chemical that has a steady level in the blood, and is freely
filtered but neither reabsorbed nor secreted by the kidneys.
 The rate therefore measured is the quantity of the substance in
the urine that originated from a calculable volume of blood. The
GFR is typically recorded in units of volume per time, e.g.
milliliters per minute ml/min. Compare to filtration fraction.
 There are several different techniques used to calculate or
estimate the glomerular filtration rate . It is also theoretically
possible to calculate GFR using the Starling equation.
Example:
 A person has a plasma creatinine
concentration of 0.01 mg/ml and in 1 hour
produces 60ml of urine with a creatinine
concentration of 1.25 mg/mL (Creatinine
clearence).
Urine formation, please see
http://www.nsbri.org/HumanPhysSpace/focus4/ep-urine.html
ACTIVE TRANSPORT
 Active transport is the mediated process of
moving particles across a biological membrane
against a concentration gradient.
 If the process uses chemical energy, such as
from adenosine triphosphate (ATP), it is termed
primary active transport.
 Secondary active transport involves the use of
an electrochemical gradient. Active transport
uses energy, unlike passive transport, which
does not use any energy.
 Specialized trans-membrane proteins recognize the
substance and allows it access (or, in the case of
secondary transport, expend energy on forcing it) to
cross the membrane when it otherwise would not, either
because it is one to which the lipid bilayer of the
membrane is impermeable or because it is moved
against the concentration gradient.

 The last case, known as primary active transport, and

the proteins involved in it as pumps, uses the chemical
energy of, usually, ATP.
 The other cases, which usually derive their energy
through exploitation of an electrochemical gradient, are
known as secondary active transport and involve pore-
forming proteins that form channels through the cell
membrane.
Antiport, symport
 Sometimes one substance is transported in one
direction at the same time as another substance is
being cotransported in the other direction. This is called
antiport.
 Symport is the name if two substrates are being
transported in the same direction across the
membrane.
 Antiport and symport are associated with
secondary active transport, meaning that one of the
two substances are transported against their
concentration gradient utilizing the energy derived from
the transport of the second substance (mostly Na+, K+
or H+) down its concentration gradient.
 http://www.youtube.com/watch?v=JGF6ry0
SWPs
Comparing Facilitated Diffusion and
Active Transport

 Transport of solutes across cell membranes by protein carriers

can occur in one of two ways:• The solute can move "downhill,"
from regions of higher to lower concentration, relying on the
specificity of the protein carrier to pass through the membrane.
This process is called passive transport or facilitated diffusion, and
does not require energy.

 The solute can move "uphill," from regions of lower to higher

concentration. This process is called active transport, and requires
some form of chemical energy. Examples of active transport
include when sodium is transported out of the cell and potassium
into the cell by the sodium-potassium pump. Active transport often
takes place in the internal lining of the small intestine.
Comparing Facilitated Diffusion and
Active Transport

The transport process a cell uses depends on its specific needs. For example, red
blood cells rely on facilitated diffusion to move glucose across membranes,
whereas intestinal epithelial cells use active transport to take in glucose from the
gut. Facilitated diffusion is effective for red blood cells because the concentration
of glucose in the blood is stable and higher than the cellular concentration.
On the other hand, active transport is needed in the gut because there are large
fluctuations of glucose concentration as a result of eating.
Active transport is responsible for the well-established observation
that cells contain relatively high concentrations of potassium ions
but low concentrations of sodium ions.
The mechanism responsible for this is the sodium-potassium pump
which moves these two ions in opposite directions across the
plasma membrane. This pump is an enzyme located in the
plasma membrane
 Export of sodium from the cell provides the driving force
for several secondary active transporters
membrane transport proteins, which import glucose,
amino acids and other nutrients into the cell by use of the
sodium gradient.

 Another important task of the Na+-K+ pump is to provide

an Na+ gradient that is used by certain carrier processes.
In the gut, for example, sodium is transported out of the
reabsorbing cell on the blood side via the Na+-K+ pump,
whereas, on the reabsorbing side, the Na+-Glucose
symporter uses the created Na+ gradient as a source of
energy to import both Na+ and Glucose, which is far
more efficient than simple diffusion. Similar processes
are located in the renal tubular system.
Na K pump
+ +
Controlling cell volume
 One of the important functions of Na+-K+ pump is to maintain the
volume of the cell.
 Inside the cell there are a large number of proteins and other organic
compounds that cannot escape from the cell. Most, being negatively
charged, collect around them a large number of positive ions.
 All these substances tend to cause the osmosis of water into the cell
which, unless checked, can cause the cell to swell up and lyse.
 The Na+-K+ pump is a mechanism to prevent this. The pump
transports 3 Na+ ions out of the cell and in exchange takes 2 K+ ions
into the cell.
 As the membrane is far less permeable to Na+ ions than K+ ions the
sodium ions have a tendency to stay there. This represents a
continual net loss of ions out of the cell. The opposing osmotic
tendency that results operates to drive the water molecules out of the
cells. Furthermore, when the cell begins to swell, this automatically
activates the Na+-K+ pump, which moves still more ions to the
exterior.
Mechanism
 The pump, with bound ATP, binds 3 intracellular Na+ ions.
 ATP is hydrolyzed, leading to phosphorylation of the pump at a
highly conserved aspartate residue and subsequent release of
ADP.
 A conformational change in the pump exposes the Na+ ions to the
outside. The phosphorylated form of the pump has a low affinity
for Na+ ions, so they are released.
 The pump binds 2 extracellular K+ ions. This causes the
dephosphorylation of the pump, reverting it to its previous
conformational state, transporting the K+ ions into the cell.
 The unphosphorylated form of the pump has a higher affinity for
Na+ ions than K+ ions, so the two bound K+ ions are released.
ATP binds, and the process starts again
 Exogenous
 The Na+-K+-ATPase can be pharmacologically modified by
administrating drugs exogenously.
 For instance, Na+-K+- ATPase found in the membrane of heart cells
is an important target of cardiac glycosides (for example digoxin
and ouabain), inotropic drugs used to improve heart performance
by increasing its force of contraction.
 Contraction of any muscle is dependent on a 100- to 10,000-times
higher than resting intracellular Ca2+ concentration, which, as soon
as it is put back again on its normal level by a carrier enzyme in the
plasma membrane, and a calcium pump in sarcoplasmic reticulum,
muscle relaxes.
 Since this carrier enzyme (Na+-Ca2+ translocator) uses the Na
gradient generated by the Na+-K+ pump to remove Ca2+ from the
intracellular space, slowing down the Na+-K+ pump results in a
permanently-higher Ca2+ level in the muscle, which will eventually
lead to stronger contractions.
 Na+/K+- ATPase was discovered by Jens
Christian Skou in 1957 while working as
assistant professor at the Department of
Physiology, University of Aarhus,
Denmark. He published his work in 1957.
 In 1997, he received the Nobel Prize in
Chemistry "for the first discovery of an ion-
transporting enzyme, Na+- K+ -ATPase.