Fluid compartments, Intracellular

compartments
Vino Dorsamy
 References
• Naish Chapter 1 pp 10 -13; Chapter 2 pp 55-
57, 62-65
• Guyton Chapter 2 p 18-19; Chapter 25 pp
285-293
• Tokarev, A.A., Alfonso, A., Segev, N., 2000. Overview
of Intracellular Compartments and Trafficking
Pathways [WWW Document]. URL
http://www.ncbi.nlm.nih.gov/books/NBK7286/?rep
ort=printable (accessed 3.17.13).
• Videos: VCell: Protein trafficking and Protein
Modification
 Objectives
• Body fluid Compartments
• Tonicity
– Solute contents
• Fluid intake and output
– Vasopressin
– Dehydration
– Water intoxication
– Oedema
 Intracellular compartments
• Endocytic /exocytic pathways
• Trafficking between Golgi and endosomes
• Transcytosis
• Late endosome to plasma membrane
• Regulated receptor endocytosis
• Autophagy
• Compartment dynamics
 Body fluids and compartments
• Human body contains large amounts of water in
different compartments
• It is essential that correct amounts of fluids at
correct concentrations are maintained in the
different compartments for all biochemical
reactions to take place
• All substances require water to diffuse through over
short distances, or in bulk flow through blood over
longer distances
 Different compartments

• Total body water (TBW) is 60% - varies with age

and weight
• Divided into 2 main compartments –
Extracellular fluid (ecf) and intracellular fluid(icf)
• icf is 2/3 of TBW and ecf is 1/3 TBW = 14L
• Transcellular compartment –very small
compartment contains synovial, peritoneal,
pericardial and cerebrospinal- fluids
 Intracellular fluid compartment
• Intracellular fluid inside 100 trillion cells of
the body
• Concentration of substances within cells very
similar from cell to cell
• Therefore intracellular compartment
considered one large fluid compartment
 Extracellular fluid compartment
• All fluid outside of cells
• Made up of two major compartments interstitial
fluid (isf) and plasma
• isf is ¾ ecf or 15% TBW
– It surrounds the cells but is outside of the blood
vessels
– Contains very little protein and few cells in
suspension
• Blood plasma is ¼ of ecf or 5% TBW
– Fluid inside blood vessels
– Carries other components of blood eg red and
white cells, and plasma proteins in suspension
throughout the body
– Total blood volume is 5.5L in adult 70kg person
– 3.5 L is plasma and most of the rest is red blood
cells
– The ratio of red blood cells to volume of whole
blood is called haematocrit usually between 0.36
and 0.53
• All compartments are separated by
semipermeable membranes
 Fluid movements
Remember…

• Movement of water is determined by hydrostatic pressure and osmotic

pressure
• Hydrostatic pressure from pumping action of the heart
• Osmotic pressure by concentration of solute particles
• Water moves form dilute to concentrated solutions
• The more solutes there are the greater the pull on the water molecules
• Osmolarity is determined by the number of osmotically active particles per
litre
• Normal body fluid osmolarity is 282 mOsm/L –read up on osmolarity and
osmotic pressure and osmotic equilibrium in Guyton pp289-301 very impt!
 Tonicity
• Tonicity refers to the effect of a solution of a cell
• An impermeant solute is one that cannot cross a
cell membrane
• 1 mOsm of impermeant solute can cause a
19.3mmHg pressure exertion across the membrane
so if you place a cell (282mOsm/L) in pure water an
osmotic pressure of 5400mmHg can develop across
the cell membrane
• Effects of different concentrations of impermeant solutions on cell volume
• If it is placed in a 282mOsm solution the cells will not swell or shrink as the
concentration in and out of cell is equal - isotonic solution
– Eg. 0.9 percent of Sodium chloride or 5% glucose solution
– Important for infusion into blood without upsetting osmotic equilibrium
between intra-and extracellular fluids
• In a hypotonic solution (<282mOsm/L) water will diffuse into cells causing it
to swell – this will continue diluting the icf and concentrating the ecf until
equilibrium is reached
• If cells placed in a hypertonic solution water will flow out of the cell into the
extracellular fluid causing the cell to shrink. This will continue until the two
solutions reach equilibrium
– NaCl solutions 0.9 % are hypertonic
 Fluid intake and output

• Amount of water needed varies – climate and activity

• Water intake via foods and 10% by metabolism
• Water is lost via urine, faeces and sweat, and evaporation via lungs
and skin –also varies based on climate and activity
• About 300mL of urine needs to be produced to rid body of
nitrogenous wastes
• Fluid intake is controlled by sensation of thirst –stimulated by
increase in osmolarity sensed by hypothalamus in brain or a fall in
blood volume
• Immediate cessation of thirst occurs after drinking to prevent
excessive consumption –this encourages small frequent drinks of
water until fluid balance is restored
• Salty food stimulate thirst – salty popcorn
 Water loss in kidneys
• Kidneys filter the plasma to form urine
• Hydrostatic pressure of blood drives the
process – and blood flow to kidneys
• If blood volume is low then less filtrate is
formed and if blood volume is high then
more filtrate is produced
• Large volumes of filtrate is produced -
120ml/min ~to170 L/day
• Large amount of water recovered in the
tubules by transport processes and urine
passes into collecting ducts into the bladder
 Vasopressin
• Volume of urine is controlled by hormone vasopressin
• Aka antidiuretic hormone (ADH) released from posterior
pituitary
• Stimulated by osmoreceptors in hypothalamus in
response to osmolarity of blood
• Also secreted in response to fall in blood pressure –sensed
by pressure receptors in heart and large blood vessels
• 75% of water recovered by osmosis in proximal tubules
• Final urine concentration determined by latter part of
tubules
– When urine needs to be concentrated vasopressin released
and acts on cells in distal tubule and collecting ducts to
increase permeability to water
– Tissue surrounding ducts is hyperosmotic compared to dilute
urine therefore urine passes back into tissue and blood
– No vasopressin release in high blood pressure or hypo-osmotic
blood –dilute urine produced
 Dehydration
• If water intake does not cover water losses
dehydration occurs
• Water moves out of cells into isf and cells shrink
– Dry mouth, thirst, dry skin and low volume of urine
leading to weight loss, fever and confusion
– Concentration of blood also increases workload of
heart and leads to increased blood viscosity which
may lead to blood clots
• Excess urine production in diabetes
– in diabetes mellitus excess sugar in urine increases
osmolarity of urine and prevents water reabsorption
– In diabetes insipidus <40 L urine /day produced due
to failure of vasopressin release or failure of kidney
to respond to vasopressin
 Oedema
• Swelling of tissues due to accumulation of fluid
is called oedema
• Has many different causes
• Large quantities of fluid exchanged between
compartments
• Hydrostatic pressure favours fluid leaving blood
vessels over colloid osmotic pressure
• Although fluid is returned to the circulation
under normal circumstances if there is a small
amount retained by the tissues oedema occurs
• Eg feet after a long day in shoes that fit
Intracellular compartments and trafficking
pathways
• All cells surrounded by a barrier between inside and
the environment – membrane
• Prokaryotic cells use their only membrane - plasma
membrane to carry out membrane attached functions
in specialised patches of the membrane
• Specialisation in eukaryotic cells make them more
advanced and allow them to attain a larger size: 1000-
10000 fold increase
• Specialised membrane bound intracellular
compartments carry out specific functions eg. Nucleus
for DNA replication and mitochondria for respiration
• Compartmentalisation allows for greater size,
complexity and efficiency but creates a problem
• How do you communicate between the cellular
compartments?
 Vesicular transport
• Major part of communication is vesicular
transport
• Cargo loaded vesicles form at a donor
compartment with the help of specific coat
and adaptor proteins (COPI and II, clathrin)
• Vesicles then target to appropriate acceptor
compartment to which they attach by tethers
(SNAREs)
• Vesicular transport enables proteins in
membrane bound vesicles to move between
cell compartments including the outer cell
membrane
 Two major pathways
• Shuttles material outward or inward
• Exocytic pathway
– Proteins synthesized in the cytoplasm
translocated into Endoplasmic reticulum (ER)
– Rough ER is where all secreted proteins are made
and all proteins resident in compartments
connected by vesicular transport
– ER is where most lipids in cells made
– From ER membranous vesicles shuttle cargo to
Golgi apparatus
– ER cargo enters via the cis cisterna and moves
through the medial and trans cisternae
– In trans cisternae proteins destined fro secretion
or for incorporation with the PM are packed into
secretory vesicles that fuse with the PM
– This occurs either constitutively (to replace used
or lost membrane) or regulatory, as a response to
an external signal
– Golgi is also a major sorting compartment as
cargo is also transported to endosomes and
lysosomes and back to ER
• Endocytic pathway
– Proteins and membrane are internalised from the
environment via endosomes (early and late) to the
lysosome
– Membrane receptors internalised via clathrin coated
vesicles
– Proteins and viruses are internalised by caviolar or raft
dependent routes
– Dependent on GTPase dynamin for fission of the
forming PM vesicle
– Fluid phase cargo may also enter via a dynamin
independent route
– Popular pathway is clathrin coated vesicles through
early and late endosomes to lysosomes
– In the first set of endosomes (sorting) cargo is sorted
for recycling back to the PM (or Golgi) via recycling
endosomes or to the lysosome via late endosomes
– Lysosome is major degradation sit e for internalised
material and cellular membrane proteins
• Both pathways mediated by vesicular transport
machinery and there is cross talk between both
pathways
– Bidirectional movement between Golgi and
endosomes, transport form one side of a polarised cell
to the other and secretion of materials from late
endosomes
 Trafficking between the Golgi and endosomes
• All proteins/lipids destined for function in
any compartment connected by vesicular
transport need to be translocated to ER
first, there needs to be a pathway to
transport newly synthesized endosomal and
lysosomal proteins and lipids to endocytic
compartments
• These proteins are labelled with mannose-
6-phosphate (M6P) in the Golgi.
• M6P cargo then sorted by M6P receptors in
the trans Golgi into vesicles that are
targeted endocytic compartments
• Lower pH in endosomes causes
disassociation of the M6P receptor from the
cargo and the receptor is recycled back from
endosomes back to Golgi
 Transcytosis
• Polarisied cells such as epithelial cells and neurons
contain distinct functional PM domains: apical and
basolateral and somatodentritic and axonal,
respectively
• Polarised cells use endocytic pathway to shuttle
cargo between distinct PM domains
• Cargo is internalised from PM on one side of the
cell eg apical side that faces the lumen of organs
• Cargo first delivered to early endosomes can be
shuttled via common set of late endosomes and
then through basolateral early endosomes to the
PM on the basolateral side
• Thus transcytosis can selectively move material
through cells across tissue barriers eg epithelium
lining the intestines
• Exocytic machinery like tethering complexes and
SNAREs are required for this process
 Late Endosome to Plasma Membrane
• Transported macromolecules of late endocytic
compartment is redirected to the PM and secreted
inside small vesicles – exosomes
• Multivesicular bodies (MVBs) are late endosomes
that contain internal membrane surrounded cargo
• Normally fuse with lysosomes but can fuse with PM
and secrete exosomes to external environment
• Important for communication between cells and
used for secretion of components into blood stream
and signalling
• Plays a role in spreading of infectious agents eg HIV
can hijack this route to be released from cells
 Further reading AKA Homework
• Regulated trafficking
• Regulated receptor
endocytosis
• Autophagy
Videos