Cell 1 & 2

Assoc Prof Rosnah Ismail

Department of Physiology
Faculty of Medicine
22 & 27 July 2010
Introduction
 Lecture Objectives

1. describe basic cell structure

2. name cellular organelles and their functions
3. describe the structure of cell membrane
4. explain various transport processes across cell
membrane
Passive, active, special
Osmosis; osmolarity and calculation for it
5. describe the distribution of ions between
extracellular and intracellular fluids (Na+ & K+)
6. define Resting Membrane Potential and explain how
it is generated
 Cell Structure
Cells differ from one type to another
epithelial, muscle, nerve etc

Epithelial cell Muscle cells Bone cells Nerve cell

BUT all cells have one
BASIC structure:
Membrane; cell membrane,
membrane of organelles

Cytoplasm; mainly water,

electrolytes and proteins

Organelles; nucleus,
mitochondria, Golgi apparatus
etc

Size of cells: 10-20m

 Cell Membrane
Functions:
1. Regulates movement of substances in and out of cells
2. Senses neurotransmitter/hormone at the surface of cells
(receptor)
3. Connects adjacent cells
4. Binds various proteins involved in generation of force
and transfer of force
5. Separates the extracellular fluid from intracellular fluid
Characteristics of cell membrane

Semi permeable/ selective permeability

compare permeability towards polar molecule to
that of non-polar molecules??
small molecules compared to big ones??

Causes differences between ICF and ECF

Intracellular Fluid (ICF) versus
Extracellular Fluid (ECF)

ICF

ECF
 Cell Membrane
Membrane thickness: 6-10nm
Structure: (fluid-mosaic model)
consists of: 1. phospholipid bilayer
2. protein
3.cholesterol

Phospholipid: Phosphate:
hydrophilic
Amphiphatic molecule
Lipid:
hydrophobic
 ECF

ICF

No chemical bonds between phospholipid molecules

• very fluid
• cholesterol stabilises the membrane
 Cell Membrane Proteins
Mostly glycoproteins

• integral (intrinsic) proteins

protrude through membrane
channels, carrier, membrane enzyme,
receptors

• peripheral (extrinsic) protein

attached to one surface of membrane, onto integral
protein, does not penetrate membrane
 peripheral
protein
Integral protein

Integral protein
Junctions between cells
 Transport across Membrane
1. Passive
Simple Diffusion
Facilitated Diffusion
Osmosis
2. Active : Primary, Secondary
3. Special processes:
Filtration
Endocytosis
Exocytosis
 Passive Transport
No energy requirement
• Movement of solutes down gradient;
concentration, electrical
due to random motion (Brownian movement)
• Movement of solutes through
lipid bilayer
channels
carrier (facilitated diffusion)
 Passive Transport
Factors affecting diffusion
1. Concentration gradient
2. Electrical gradient
3. Size of molecules
4. Lipid solubility
5. Surface area
6. Thickness of membrane
7. Temperature
Q = rate for diffusion
D = diffusion coefficient (depends on
Q = D(C1-C2) properties of the molecule, membrane
and temperature)
C1-C2 = concentration gradient
 Simple Diffusion

Movement of solutes is from

HIGH conc LOW conc
 Facilitated Diffusion

Solute binds to specific carrier on membrane

· carrier undergoes conformational change
· solute transported to opposite surface
· solute unbound from carrier
· carrier back to original conformation
Example: transport of glucose into red blood cell
 Factors affecting rate of
facilitated diffusion
• Same factors as for simple diffusion
• 2 additional factors
Saturation
competitive inhibition
 Characteristics of carrier-mediated transport:
Shows Saturation
- number of carrier proteins available
- the rate at which binding/unbinding
occurs

Shows Competitive/ non- Normal

competitive inhibition

Rate of transport
- presence of two or more types of
molecule transported by same
carrier protein
- they compete/ bind to same carrier

Concentration of transported molecule

 Osmosis
Definition: movement of solvent (water) down its
concentration gradient across a membrane selectively
permeable to it
passive process

• water is a polar molecule

• size 0.3nm
• crosses lipid bilayer of membrane or
through water channels (aquaporin)
 Osmosis
Visualise 2 solutions with different concentrations
separated by a membrane permeable
i. to both water and solutes
ii. to water only

solution with low solute conc, has high water conc

solution with high solute conc, has low water conc

What will happen?

 Concept of Osmotic Pressure

Osmotic pressure: the pressure required to stop

movement of water (osmosis) is the osmotic pressure
of the solution
Unit = mmHg

solution with high solute concentration

high osmotic pressure

solution with low solute concentration

low osmotic pressure
OSMOSIS
Osmotic Pressure
 Osmotic Pressure
Osmotic pressure depends on:

1. Number of non-penetrating solutes

per unit volume of fluid

DOES NOT depend on size/mass of solutes

the higher the number of non-penetrating solutes in a solution,
the higher the osmotic pressure

Number of solutes in a solution depends on:

i. Concentration of the molecule in a solution
(Molarity)

ii. Whether the molecule dissociates in solution

2. Temperature P= osmotic pressure
R= gas constant
van’t Hoff Law: P= nRT T= absolute temp ( K)
V n= no solutes
V= volume of solution
(true only for dilute solution)
 Terminologies
Osmolality: Concentration of solution in terms of
number of solutes per kg water.
Unit: Osmole/kg

Osmolarity: Concentration of solution in terms of

number of solutes per liter water.
Unit: Osmole/L

1 Osmole = 1000 mOsmole

(Osm) (mOsm)
Osmolarity of body fluid: 285-300 mOsm/l

ECF & ICF

285-300 mOsm/L (0.285 – 0.3 Osm/L)

equivalent to osmotic pressure of
 7.3 atmospheric pressure
(5500 mmHg)
 Osmolarity of a Solution
Number of molecules in a solution depends on the
molarity of the solution

No of Osmole = Molarity of substance X no. of freely moving

solutes that each molecule liberates in soln
For non-dissociable molecule:
e.g. soln of 0.3 M; will have 0.3 Osm since;
no. of Osm = 0.3 X 1 = 0.3 Osm
For dissociable molecule:
e.g. substance A dissociates into 3 solutes in solution
0.3 M of substance A will have 0.9 Osm since;
no. of Osm = 0.3 X 3 = 0.9 Osm
Examples:
1. Glucose does not dissociate in solution.
1 M glucose solution = 1 Osm/L
What is the osmolarity of 0.3 M/L glucose solution?
Osm = 0.3 X 1 = 0.3 Osm/L (300 mOsm/L)

2. NaCl (dissociates into 2 ions: Na+ and Cl-)

What is the osmolarity of a 1 M NaCl solution?
Osm = 1 X 2 = 2 Osm/ L
(In this solution there are: 1 Osm/L Na+ + 1 Osm/L Cl- ions)
 Some Exercises to do

Calculate the osmolarity of the following solutions:

1. 145 mM NaCl
2. 0.15 M CaCl2
3. 0.15 M Na2SO4
4. 0.90% NaCl (MW NaCl= 58.5)
Isosmotic, Hyposmotic & Hyperosmotic

compare A to B

300 300 200 isosmotic

mOsm/L mOsm/L mOsm/L
Compare A to C
hyperosmotic
A B C Compare C to A or B

hyposmotic
 Concept of Tonicity
Comparison between osmolarity of a solution to that of
plasma
Body fluid Osmolarity about 300 mOsm/l
Osmolarity ICF = Osmolarity ECF
ICF and ECF in equilibrium

 looks at the non-penetrating solutes

 considers whether there is a change in cell size
when cells are put into the solution
 Isotonic, Hypotonic, Hypertonic
Isotonic solution:
concentration of non-penetrating solutes is same as that of plasma
(body fluid), 300mOsm/L
water influx = water efflux (no net movement of water)
no change in cell size

Hypotonic solution:
concentration of non-penetrating solutes is less than that of plasma;
< 300mOsm/L
water influx > water efflux (net movement of water into cells)
cells swell

Hypertonic solution:
concentration of non-penetrating solutes is more than that of plasma,
>300 mOsm/l
water influx < water efflux (net movement of water out of cells)
cells shrink (crenation)
 300 mOsm/L
300 mOsm/L
NaCl
NaCl

300 300
mOsm/L mOsm/L

NaCl does not penetrate cell membrane

No change in cell size

NaCl solution with solute concentration of 300 mOsm/L
is isosmotic as well as isotonic compared to cell
 300 mOsm/L
urea
300 mOsm/L
urea
urea

300
mOsm/L
300
mOsm/L + 300 mOsm/L
urea
water

Urea penetrates cell membrane easily

Cells swell
This solution of urea with solute concentration of 300
mOsm/L is isosmotic but is NOT isotonic compared to cell
 Some More Exercises to do

What will happen to red blood cells if they are placed in:
1. 150 mOsm/L NaCl solution?
2. A solution containing mixture of NaCl 300 mOsm/L
and urea 150 mOsm/L?
3. 300 mOsm/L glucose solution?
Measurement of Osmolarity of a Solution

• Using the principle of freezing point depression

• Pure water freezes at 0°C
• Addition of solutes decreases the freezing point of
water, < 0°C

• KNOWN:
1 mole of an ideal solution depresses the freezing
point of water by 1.86C

• Can use this fact to measure the osmolarity of a solution

e.g if the freezing point of a solution decreases by 0.56C
osmolarity of the solution is 0.56/1.86
about 0.3 Osm/L
 Active Transport
Requires energy (ATP)
Uses carrier
Carrier has ATPase enzyme
(to hydrolyse ATP, releases energy)
Substance transported against concentration gradient
Can occur in living cells only

2 major types:
• Primary
• Secondary
 Primary Active Transport
Uses energy (ATP) directly to transport substances
against electrochemical gradient across cell membrane
Examples:
Na+,K+- ATPase (in all cells)
Ca2+-ATPase (e.g. in sarcoplasmic reticulum of muscles)
H+-K+ ATPase (cells in stomach, kidneys)
H+- ATPase (lysosome)
 Types of Transporters
Transports one type of solute: uniport
Transports 2 types of solutes in same direction: symport/co-transport
Transports 2 types of solute in opposite directions : antiport/counter-transport
Na,K- ATPase
 Secondary Active Transport
Indirect use of Energy (ATP)
Carrier has 2 binding sites for two different solutes
one for ion (Na+) and one for another solute

E.g: glucose, galactose, amino acid

in kidneys and small intestine
1. conc gradient for Na+ provides
the energy (High Na+ conc
outside, low inside due to action
of Na+-K+-ATPase pump)
2. Na+ binds to carrier because of
presence of gradient
3. Glucose binds to carrier
4. Carrier undergoes
conformational change
5. Na+ released into cytoplasm,
glucose released from binding
site into cytoplasm
6. Carrier goes back to original
conformation
7. Na+ pumped out (by Na+-K+
ATPase pump; utilising ATP) and
Na+ gradient maintained
 Special Transport Process
1. Filtration
Transport by bulk flow across a
membrane due to difference in
pressure

Both solutes and solvent cross membrane

Dependent on Effective Filtration Pressure

e.g Filtration at the renal glomerulus

Filtration of substances across capillary
endothelium into the interstitial space
 Endocytosis: Process whereby a substance
is taken up by cell
Phagocytosis
(cell eating):
solid particulate
(dead cell, bacteria)
engulfed by cell

Pinocytosis
(cell drinking) :
liquid substance
engulfed by cell
Exocytosis: secretion of substances from cell out to
the ECF
e.g. secretion of neurotransmitter from vesicle
 Introduction to Resting Membrane Potential

Selective permeability of Cell Membrane leads to different

ionic distribution between ICF and ECF:

it is relatively more permeable towards K+

(conc K+ ICF >>> K+ ECF; 4.5 mM in ECF compared to 150 mM in ICF)

it has very low permeability towards Na+

(conc Na+ ICF<<< Na+ ECF; 145 mM in ECF compared to 15 mM in ICF)

It is not permeable to proteins

(protein ICF > protein ECF)
 Na+

Na+ Na+ Na+

Na+ Na+

Na+ K+ K+
K+ K+
Na+ K+

K+ 3Na+
K+ K+ K+
K+
Na+ K+
Na+ 2K+
K+

Na+ Na+ K+
K+ K+
K+ Na+
prot- prot-
prot- prot-
Na+
Na+

Na+
Na+ Efflux K+ Na+/K+ ATPase
Na+ pump
Influx Na+

Prot cannot pass through membrane

 Resting Membrane Potential (Em)
Resting membrane potential is potential difference across
the cell membrane at rest: inside all cells is negative
1. Membrane permeability to K+ is relatively high
compared to Na+;
Em  log [Ko+]
[Ki+]
(Number of molecules involved in generation of Em is very small)

2. Negatively charged proteins cannot penetrate

membrane

3. Na+/K+ ATPase pump is electrogenic, pumping more

+ve charges out than in (3 Na+ out and 2 K+ ions in),
contributing to the negative RMP
 At rest, inside all cells is negative
Resting Membrane Potential (Em): - 10 to – 100 mV
Em of Excitable cells: - 60 to -100 mV
Em of other cells: - 10 to – 30 mV