I.

Introduction
A. The Origin and Evolution of Cells B. Cells as Experimental Models C. Tools of Cell Biology

Cell biology (formerly cytology, from

the Greek kytos, "container") is an academic discipline that studies cells
their physiological properties their structure, the organelles they contain interactions with their environment, their life cycle, division and death.

Cell basic unit of life structurally and functionally.

History:
a. Robert Hooke (1665)using his microscope discovers cells in cork b. Schleiden ; Schwann and Virchow Cell theory: 1. All organisms are composed of one or more cells 2. The cell is the structural unit of life 3. Cells can arise only by division from preexisting cells

Various cell types: shape, size, intracellular organizations, polarization Functions

Dynamic Nature of Cell

-it has the capacity to grow to reproduce become specialized ability to respond to stimuli adapt to changes in its environment

Fundamental properties shared by all cells: (conserved throughout evolution) 1. all cells employ DNA as their genetic material 2. surrounded by plasma membrane 3. use the same basic mechanisms for energy metabolism

Two Main Classes of Cells:

a. Prokaryotic cells no nucleus - simpler structure (bacteria)

b. Eukaryotic cells - contain nucleus - more complex structure(protists, fungi, plants & animals)

Fig.1.2.Average_prokaryote_cell-_en.svg(SVG file, nominally 494 402 pixels, file size: 135 KB)

Fig1.3.Diagram of a typical animal (eukaryotic) cell, showing subcellular components. Organelles: (1) nucleolus (2) nucleus (3) ribosome (4) vesicle (5) rough endoplasmic reticulum (ER) (6) Golgi apparatus (7) Cytoskeleton (8) smooth endoplasmic reticulum (9) mitochondria (10) vacuole (11) cytoplasm (12) lysosome (13) centrioles within centrosome

Fig.1.4.Diagram of a typical plant cell

Table 1: Comparison of features of prokaryotic and eukaryotic cells

Typical organisms Typical size Type of nucleus DNA RNA-/protein-synthesis Ribosomes Cytoplasmic structure Prokaryotes bacteria, archaea ~ 1-10 m nucleoid region; no real nucleus circular (usually) coupled in cytoplasm 50S+30S very few structures Eukaryotes protists, fungi, plants, animals ~ 10-100 m (sperm cells, apart from the tail, are smaller) real nucleus with double membrane linear molecules (chromosomes) with histone proteins RNA-synthesis inside the nucleus protein synthesis in cytoplasm 60S+40S highly structured by endomembranes and a cytoskeleton flagella and cilia containing microtubules; lamellipodia and filopodia containing actin one to several thousand (though some lack mitochondria) in algae and plants single cells, colonies, higher multicellular organisms with specialized cells Mitosis (fission or budding) Meiosis 1.5 107 to 5 109

Cell movement Mitochondria Chloroplasts Organization Cell division DNA content (base pairs)

flagella made of flagellin none none usually single cells Binary fission (simple division) 1 106 to 5 106

Organisms: 1. Unicellular (eg. bacteria, amoebas & yeasts) capable of independent selfreplication 2. Multicellular(eg. Humans)- composed of collection of cells w/c fxns in a coordinated manner w/ diff cells specialized to perform particular tasks.

the First Cell:

-all present day cells (both prokaryotes & eukaryotes) descended from a single ancestor. -the 1st cell is thought to have arisen at least 3.8 B years ago as a result of enclosure of selfreplicating RNA in a phospholipid membrane (RNA world hypothesis)

Present-Day Prokaryotes -divided into two groups: the archaebacteria and the eubacteria which diverged early in evolution
Eukaryotic Cells -thought to have evolved from symbiotic associations of prokaryotes (ENDOSYMBIONT THEORY)

ENDOSYMBIOSIS
A large anaerobic, heterotrophic prokaryote engulfs a small aerobic prokaryote

The aerobic endosymbiont has evolved into a mitochondrion

A portion of the plasma membrane has invaginated and evolved into a nuclear envelope and endoplasmic reticulum (primitive eukaryote) Engulfs a photosynthetic prokaryote Evolve into a chloroplast

Nonphotosynthetic protist, fungal, animal cells

Algal & plant cells

Fig.1.5. Time scale of evolution The scale indicates the approximate times at which some of the major events in the evolution of cells are thought to have occurred.

The Evolution of Metabolism:

Figure 1.6. Generation of metabolic energy Glycolysis is the anaerobic breakdown of glucose to lactic acid. Photosynthesis utilizes energy from sunlight to drive the synthesis of glucose from CO2 and H2O, with the release of O2 as a by-product. The O2 released by photosynthesis is used in oxidative metabolism, in which glucose is broken down to CO2 and H2O, releasing much more energy than is obtained from glycolysis.

Figure 1.6. Evolution of cells Present-day cells evolved from a common prokaryotic ancestor along three lines of descent, giving rise to archaebacteria, eubacteria, and eukaryotes. Mitochondria and chloroplasts originated from the endosymbiotic association of aerobic bacteria and cyanobacteria, respectively, with the ancestors of eukaryotes.

Cells as Experimental Models

E. coli

S. cerevisiae

Dictyostelium discoideum

Arabidopsis thaliana

Caenorhabditis elegans

Drosophila melanogaster

Xenopus laevis

zebrafish

House mouse

Choosing the Right Experimental Organism for the Job

Table 1.2 DNA Content of Cells

Organism Haploid DNA content (millions of base pairs)

Bacteria Mycoplasma E. coli Unicellular eukaryotes Saccharomyces cerevisiae (yeast) Dictyostelium discoideum Euglena Plants Arabidopsis thaliana Zea mays (corn) Animals Caenorhabditis elegans (nematode) Drosophila melanogaster (fruit fly) Chicken Zebrafish Mouse Human

0.6 4.6 12 70 3000 130 5000 97 180 1200 1700 3000 3000

Tools of Cell Biology

1. Light Microscopy Bright-field microscopy Phase-contrast microscopy Differential interference contrast microscopy Video-enhanced differential interference-contrast microscopy Fluorescence microscopy Confocal scanning microscopy 2. Electron microscopy Transmission electron microscopy Scanning electron microscopy

Light Microscope
earliest tool of cytologist Limit of resolution is /2= 0.20-0.35 um - To magnify an object, it uses a system of lenses to manipulate the path a light beam travels between the object being studied and the eye - Produce a maximum useful magnification of about 1000 times the original size. - Has three lenses: 1. the condenser(focuses light on the specimen 2. the objective(s) a. Low-power objective b. High-power objective c. Oil-immersion objective 3. the eyepiece (ocular)

Table 2-1 Objective Designation

Low Power High Power (high dry) Oil Immersion

Objective Magnification
10 40 100

Eyepiece Magnification
10 10 10

Total Magnification
100 400 1000

Table 2-2. Metric unit equivalents in expressing cell dimensions

Unit of length Micrometer (um) Nanometer (nm) Angstrom () Meter (m) 0.000001 10-6 0.000000001 10-9 0.0000000001 10-10 Centimeter (cm) 0.0001 10-4 0.0000001 10-7 0.00000001 10-8 Millimeter (mm) 0.001 10-3 0.000001 10-6 0.0000001 10-7 Micrometer (um) 1 0.001 10-3 0.0001 10-4 Nanometer (nm) 1000 103 1 0.1 10-1

A. Light Microscope
Terms: 1. Limit of Resolution-refers to how far apart adjacent objects must be in order to be distinguished as separate entities. (eg. LOR of microscope is 400 nm) 2. Resolving Power-expressed in terms of (the wavelength of light used to illuminate the sample) -the smaller is the limit of resolution the greater is the resolving power
Resolving Power of any microscope -a measure of its ability to discriminate between two adjacent objects. - is a function of the wavelength of light and the numerical aperture of the lens system - light microscopes (using visible light) have RP of approximately 0.25 um which means that particles of a smaller size cannot be distinguished from one another).

Fig.1-9. Resolving Power Of the Human Eye, the Light microscope and the Electron Microscope

1 um = 10-6 m (one-millionth of a meter) 1 nm = 10-9 m (one-billionth of a meter) 1000 nm = 1 um Angstrom () = 10-10 m or 0.1 nm

Type of Microscopy Bright-field

Maximum useful magnification 1000-2000

Appearance of specimen Specimens stained or unstained; bacteria generally stained and appear color of stain Generally unstained; appears brightorlightedin an otherwise dark field

Useful Applications Gross morphological features of bacteria, yeasts, molds,algae and protozoa Microorganisms that exhibit some characteristic morphological feature in the living state and in fluid suspension e.g. spirochetes

Dark-field

1000-2000

Fluorescence

1000-2000

Bright and colored; color of the fluorescent dye

Diagnostic techniques where fluorescent dye fixed to organism reveals the organismsidentity

a. Differential-interference-contrast micrograph of a mitotic yeast cell.

b. Fluorescence microscopy

c. Phase-contrast micrograph of fibroblasts in culture.

Type of Microscopy Phase-contrast

Maximum useful magnification 1000-2000

Appearance of specimen Varying degrees of darkness

Useful Applications Examination of cellular structures in living cells of the larger microorganisms, e.g yeasts, algae, protozoa and some bacteria

a) Flourescent microscope b) flourescent micrograph of chromosomes and mitotic spindle

c) Dark-field photomicrograph of Mysis d)Phase-contrast micrograph of a cheek cell

B. Electron Microscope -uses a beam of electrons controlled by a system of magnetic fields -has high resolving power thus greater magnification (can resolve objects separated by a distance of 0.003 um compared to 0.25 um of light microscope). -useful magnification is 200,000 to 400,000 -use in the examination of viruses and the ultrastucture of microbial cells. - has two types: a. Scanning electron microscopy (SEM) -employed to study the surface structure of a specimen(eg. Attachment of bacterial cells to objects) b. Transmission electron microscopy (TEM) -used to view subcellular components (even nucleic acid molecules)

Scanning electron micrograph of a flea

Transmission electron micrograph of Bacillus anthracis

Several different techniques exist to study cells: 1. Cell culture 2. Immunostaining 3. Computational Genomics 4. DNA MICROARRAYS 5. Gene knockdown 6. In situ hybridization 7. Polymerase Chain Reaction (PCR) 8. Cell Fractionation

Figure 1-7. A procedure used to make a transgenic plant.

Figure 8-63. Using cluster analysis to identify sets of genes that are coordinately regulated. Figure 1.8. Using DNA microarrays to monitor the expression of thousands of genes simultaneously.

Basic Local Alignment Search Tool (BLAST)

Figure 8-47. Results of a BLAST search. Sequence databases can be searched to find similar amino acid or nucleic acid sequences. Here a search for proteins similar to the human cell-cycle regulatory protein cdc2 (Query) locates maize cdc2 (Subject), which is 68% identical (and 82% similar) to human cdc2 in its amino acid sequence.

Thank You for Listening.