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Limits to cell size

Cell Size

growth

mitosis

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Surface Area to Volume ratio
– limits cells size To demonstrate this concept… Which has a greater surface area to volume ratio?
(provide calculations to support your answer) a) Cell 1: 1µm x 1µm x 1µm b) Cell 2: 5µm x 5µm x 5µm c) Multicellular Organism 1: 125(1µm x 1µm x 1µm) d) More than one of the above

Cell 1

Cell 2

Multicellular Organism 1

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Limits to Cell Size: Surface to Volume Ratios

*Similar to Figure 4-1

Surface Area to Volume ratio
- limits cells size
SA to V ratio is critical to cell metabolism SA (x2) = plasma membrane V (x3) = cell contents

…As cells increase in size…
• V (x3) grows proportionately more than SA (x2)
…leads to…

• Lower SA to V ratio
…leads to…

• Problematic exchange of substances between cell & environment
…affects...

• Localized [molecule] • Diffusion rate of molecules in cell
 Affects RATES of chemical reactions …Slow is not good!

How do cells cope with SA:V constraints?

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Strategies to cope with SA:V constraints
Cells divide
mitosis

Cell Size

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Strategies to cope with SA:V constraints
Growth Stops
- Cell enters G0phase
• • Withdraw from cell cycle No growth/proliferation

- Cell may become:
• Quiescent - Phase is reversible
i.e. some cells can re-enter Cell cycle to begin division e.g. some stem cells

Terminally differentiated - Non-reversible
e.g. cardiac muscle cells

Fig 19-32

Strategies to Increase Surface Area
Membrane Folding: e.g. brush border cells of intestinal epithelium

Figure 4-2

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Strategies to Increase Surface Area
Membrane Folding – also seen in some prokaryotes
e.g. Anabaena azollae (type of Cyanobacteria)

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Figure 11-4

Strategies to Increase Surface Area
Membrane Folding – not limited to the plasma membrane in eukaryotes
e.g. endoplasmic reticulum e.g. thylakoid membrane in chloroplast

Other Strategies Cells use to Cope with SA:V Constraints?

Which has the greatest surface area to volume ratio? a) Cell 1: 1µm x 1µm x 1µm

b) Cell 2: 5µm x 5µm x 5µm
c) Organism 3: 125(1µmxµm1xµm1) d) Cell 4: 0.125µm x 5µm x 200µm
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e) More than one of the above

200

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0.125

Strategies to Increase Surface Area Long, thin cells – greater surface area:volume

Muscle fiber

Neuron
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What about egg cells? Do egg cells break the SA:V rule?

Figure 1. Comparisonof egg sizes. Ostrich egg (right), compared to chicken egg (lower left) and quail eggs (upper left).
Photo by Rainer Zenz.

Do egg cells break the SA:V rule?
• Although some egg cells can be quite large…
– Ostrich eggs are several centimeters long!

• …Metabolically inactive • …Mostly lipids (fats), storage materials • …Actual viable ‘cell’ component is very small

Strategies to cope with SA:V constraints: Active Transport
• Transportation of ‘cargo’ by specialized carrier proteins

• May involve:
• Cytoskeleton
- Motor proteins + filamentous tracks

• Carrier Proteins
- Transport across membranes

• ACTIVE process = Expenditure of E by the cell

Strategies to cope with SA:V constraints: Cytoplasmic Streaming
• Bulk movement of cytoplasm
– Involves microfilaments

• Active process (E expenditure)

http://bio1151.nicerweb.com/med/Vid/Campbell7e/CytoplasmicStream-V.swf

Strategies to cope with SA:V constraints: Multicellularity
• SA:V sets an upper limit on cell size • The only way to get larger is for cells to cooperate • Driving force behind evolution of multicellularity?
*Similar to Figure 4-1
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- Volume stays the same - Surface Area increases

Question to ponder: What factors make it possible for eukaryotic cells to be so much larger than prokaryotic cells?

Ave. Size of Prokaryotic Cell ~1-5 um diameter Ave. Size of Eukaryotic Cell ~10 – 100 um diameter
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“Tour of the Cell”

‘Typical’ Prokaryotic cell
• Ave. Size of Prokaryotic Cell = ~1-5 um diameter

• Relatively ‘Simple’ Organization:
- Cell wall: protective layer of carbohydrate surrounding plasma membrane - Plasma membrane

Motility structure (e.g. flagella)

- DNA (not enclosed) – nucleoid region
-Some species have plasmids

- May/not have motility structure

Electron micrograph of cross section throughE. coli.

Eukaryotic Cell
e.g. ‘typical’ Animal cell
ENDOPLASMIC RETICULUM (ER)

Nuclear envelope Nucleolus Chromatin NUCLEUS

Rough ER Flagelium Centrosome
(with centrioles)

Smooth ER

Plasma membrane

CYTOSKELETON
Microfilaments Intermediatefilaments Microtubules

Ribosomes

Microvilli
Golgi apparatus Peroxisome Mitochondrion Lysosome Similar to Fig 4-5
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Eukaryotic Cell
e.g. typical Plant cell
Nuclear envelope NUCLEUS

Nucleolus Chromatin
Centrosome

Rough endoplasmic reticulum

Smooth endoplasmic reticulum

Ribosomes (small brwon dots)

Central vacuole

Golgi apparatus

Tonoplast Microfilaments Intermediate filaments Microtubules CYTOSKELETON

Mitochondrion Peroxisome Plasma membrane Cell wall Plasmodesmata Wall of adjacentcell Chloroplast

Similar to Fig 4-6

In plant cells but notanimal cells: Chloroplasts Central vacuole and tonoplast Cell wall Plasmodesmata

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Before we discuss cells… a few cell/molecular biology techniques…

Cell Fractionation
• Separation of a cell structures/components • Two phases 1. Homogenization
• lysing cells open • chemicals, enzymes, or sound waves

2. Centrifugation
Homogenize tissue
Mouse liver

Centrifugation
• Use of centrifugal force to separate mixture(s)
– Fixed-angle – Swinging-bucket

• Separation based on density & size
– “Pellet” = Substance(s) that sediment at bottom – “Supernatant” or “Super” = remaining liquid – Sometimes have an “Interface” – region between two phases
Figure 12A-1

Sedimentation coefficient (S)
= Measure of how rapidly a sample sediments when subjected to centrifugal force

Similar to Figure12A-2

S values ____________ from left to right.
a)Increase b)Decrease

S values ____________ from left to right.
a)Increase b)Decrease

• Larger/denser objects sediment more quickly (large S value) • Smaller/ less dense objects sediment less quickly (smaller S value)

Sedimentation coefficient (S)

Figure 12A-3

Tour of the Cell…

Keep in mind as we take our tour…

• Cells are DYNAMIC SYSTEMS!!!
Cells can adjust structures to meet changing needs
– e.g. adjust # of mitochondria to meet metabolic needs

Subcellular structures interact with one another
– i.e. they are not isolated entities

Cytoplasm
- Internal contents of cell - Contains: • Cytosol
~semi-fluid material •Organelles (eukaryotes)

• Subcellular structures
e.g. ribosomes

Plasma Membrane
• 4-8nm (40-80 Å) thick layer of lipids + protein •Boundary between cell and external environment
- defines ‘cell’ - regulates movement in/out - mediates communication with external environment (e.g. receptors)

•Fluid Mosaic
- many components (mosaic) - flexible, dynamic structure; not static

*More about this later (Ch. 7)…

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Extracellular Matrix (ECM)
• Region outside of cell
(“extra” cellular)

• Made of various proteins/polysaccharides (varies by species)
– e.g. collagen (fibrous protein); e.g. cellulose (polysaccharide)

• Linked to cell via Plasma membrane components
– e.g. integrins (intermembrane proteins)

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Functions of the ECM:
Include: – Support/Structure
• e.g. Cell Walls in plants (…more on this in a bit…) Bone matrix Protection against osmotic pressure changes

– Adhesion/anchorage to surrounding medium
• e.g. tissue formation

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ECM: Plant Cell Walls
– ECM of plant cells – Made of cellulose fibers embedded in other polysaccharides and protein
Cell Wall

– Functions
• mechanical strength • growth/development • protection

Fig 17-24

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ECM: Prokaryotic Cell Walls
Bacteria
Peptidoglycan:
• NAM-NAG polysaccharide • Cross-linked with peptides
(short amino acid chains)

Archaea
4 known variations: a. Sulfated polysaccharides b. Glycoproteins stabilized by Na+ c. S-layers – protein chain mail d. Pseudomurein (shown below)
• NAG – NAT polysaccharide • Cross-linked with peptides

N- Acetylmuramic acid (NAM) N- Acetylglucosamine (NAG)

N-Acetyltalosaminuronic acid (NAT)

Gram stain
• Tool to ID bacteria based on cell wall characteristics
 Cells stained with violet dye  Rinse with alcohol  Stain again with counter-stain (usually red dye)
Lipopolysaccharide Peptidoglycan layer Plasma membrane Protein Grampositive bacteria 20 m Gramnegative bacteria

Cell wall

Outer membrane Cell wall Peptidoglycan layer Plasma membrane
Protein

(a)

Gram-positive: -Simple cell walls - cell walls have large amount of peptidoglycan - traps violet dye in the cytoplasm - alcohol rinse does not remove the violet dye, which masks the added red dye. - Cells appear violet after counter-stain is added

(b)

Gram-negative: -Complex cell walls with less peptidoglycan - Cell wall located in layer between -PM and an outer membrane - outer membrane also has lipopolysaccharides -violet dye is easily rinsed from the cytoplasm, - cells appears pink/red after counter-stain is added

Pathogenic Prokaryotic Species
• Can be Gram+or Gram- ; also Gram variable
• Examples of Pathogenic Gram+
• Clostridium botulinum (Botulism) • Bacillus anthracis (Anthrax) • Streptococcus pneumoniae (bacterial pneumonia, bacterial meningitis)

• Examples of Pathogenic Gram• Yersinia pestis (Plague) • Bordetella pertussis (Whooping cough) • Chlamydia tachomatis (STD)

• Examples of Pathogenic Gram variable
• Mycobacterium tuberculosis (Tuberculosis) • M. leprae (Leprosy)

Gram Stain
• Used as a quick diagnostic tool • Examples:
– Classification of new species
• Based on CW characteristics • Also preserves shape (e.g. spiral, rods)

– To confirm/rule out bacterial infection
• Swabs taken from wounds, joint fluids, CSF, etc.
(see recent example on next slide)

– Assessing bacterial contamination of tissue cultures – Environmental/Industrial contamination
• Natural disasters • Milk production

Predictive value of superficial cultures to anticipate bloodstream infection.
Bouza E1, Rojas L2, Guembe M3, Marín M2, Anaya F4, Luño J4, López JM4, Muñoz P5; on behalf of the COCADI Study Group.

Abstract We performed a prospective study in patients with tunneled catheters to assess the validity of Gram stain and superficial culture for anticipating catheter exit-site infection and hemodialysis catheterrelated bloodstream infection. The sensitivity and negative predictive value were high, and we succeeded in identifying a subpopulation at low risk of infection. Diagn Microbiol Infect Dis. 2013 Dec 17. pii: S0732-8893(13)00650-0. doi: 10.1016/j.diagmicrobio.2013.12.008. [Epub ahead of print] Copyright © 2013 Elsevier Inc. All rights reserved.

Prokaryotic Cell Wall-Synthesis Inhibitors
• Certain antibiotics work by inhibiting cell wall biosynthesis • Compromise integrity of the CW Makes bacteria susceptible to osmotic pressures With weakened CWs, they will generally lyse • Examples:

- Penicillin
• Interferes with peptide synthesis and peptidoglycan cross-linking
Penicillium fungus (DIC image)

– Fosfomycin
• Interferes with peptidoglycan biosynthesis
Brightfieldimage of Streptomyces fradiae

Continue our “Tour of the Cell”…

Nucleus
• Stores DNA = CONTROL CENTER • Large organelle
~5-6um diameter ~10% total cell volume

• Surrounded by Nuclear Envelope (NE)
- Double membrane layer - Supported by Nuclear Lamina (…more in a bit…) - Punctured at intervals by Nuclear Pores (…more in a bit…)
Regulated openings through nuclear envelope - Control movement of substances in/out of nucleus

Nuclear Pores
• Regulated openings through Nuclear Envelope
• Openings controlled by Nuclear Pore Complex (NPC) • NPC controls movement of substances in/out of Nucleus
Q: What sorts of molecules are moving in/out of nucleus?

Nuclear Lamina
• Network of intermediate filaments (components of the cytoskeleton)

CLSM of nuclear lamina (green) & DNA (red) from mousenucleus
Source:http://medicalphysicsweb.org/cw s/article/opinion/34548/1/SIM_1006

• Lie just beneath inner layer of Nuclear Envelope
• Scaffolding that supports nuclear structure

• Thought to also play a role in chromatin organization

Diagram source: http://www.goldmanlab.northwestern.edu/index.htm

Nucleolus
• Region within the Nucleus • Clustered regions of ribosomal RNA genes surrounded by specific RNAs & proteins • Site of ribosomal subunit synthesis • Q: How do ribosomal subunits exit the nucleus? Q: What happens after they leave the nucleus?

Nucleus

(SEM)

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Ribosome
• Found in all cell types (pro/eukaryote),and certain organelles
• Not an organelle

25 – 30 nm

Type of microscopy used?

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The process by which genetic information coded in messenger RNA directs the formation of a polypeptide sequence is called____________.
a) b) c) d) e) Transcription Transformation Translation Transgression None of the above

Prokaryotic vs. Eukaryotic Ribosomes • Ribosomes are RNP (ribonucleoprotein) complexes
Q: What is an RNP complex?? A: Complex of RNA and protein
Prokaryote

• Sizes of ribosomes differ between prokaryotes & eukaryotes

Eukaryote