Cytology - Part III

The Prokaryotic Cell Bacteria and other prokaryotes are monerans, a diverse kingdom of small usually single-celled organisms, whose origins can be traced to the most ancient forms of life on earth. If youre not a prokaryote like the bacteria you must be a eukaryotic organism such as the protista, fungi, plants, or animals. How life came to be divided into these two fundamental groups is the subject of the endosymbiotic theory.(3) The main parts of the prokaryotic cell are diagramed below in figure 3.

Click on Fig. 3 above to see an electron micrograph of Bacillus megaterium. Prokaryotes like most organisms fall into two general categories based on how they obtain their energy. For example:

some are autotrophs: Examples: chemosynthetic or photosynthetic. both obtain their food by abiotic means relying on energy of inorganic molecules or sunlight. These organisms are classified as producers. others are heterotrophs: Examples: decomposers or parasites (disease causing microorganisms. Organisms which rely on the metabolism of other living things are called consumers

Cell size for prokaryotes, with the exception of the cyanobacteria, is about 10 times less than representative eukaryotic cells, or from 1 to 10 micrometers.

The cell wall of prokaryotic cells is made of proteoglycans, molecules which are a combination of carbohydrates and protein. What are some functions of proteoglycans? In addition to the cell wall, many bacteria produce a sheath while others have a slimy protective capsule. Within the cytoplasm of prokaryotes, surrounded by a single cell membrane, all the metabolic processes such as protein synthesis, respiration, replication, etc. take place. The genetic material of bacteria, a single circular chromosome, is found in an unbounded area of the cytoplasm called the nucleoid. Many bacteria have additional genes located in small circular molecules of DNA called plasmids. Plasmids and the genes they contain may be easily transferred from one bacteria to another, possibly even between species. The flagellum found in most aquatic and soil bacteria provide movement by the unique process of spinning on an axis like a propeller. See Figure 4 below.

Many bacteria have pili. There are two varieties of pili.

One type of pili is used by related bacteria to exchange genetic material -- a process called conjugation. A second variety of pili enable bacteria to stick to their host or substrate. This ability greatly increases the chances of infection.

Modified July 8, 2005

Cytology - Part IV
The Eukaryotic Cell

Visit the University of Virginia to examine a typical animal cell or interact with a typical plant cell prepared by Ross Koning. The Eukaryotes comprise the kingdoms animalia, plantae, fungi, and protista The following table indicates whether each structure is found in plants only, animals only, neither plants or animals, or can be found in both. (click on the terms to find out more about each one.) a) microtubules b) plastids c) lysosome d) pili e) centrioles f) nucleolus g) chloroplast h) plasmid i) j) *vacuoles peroxisome both plants animals neither animals both plants neither both both

k) plasmodesmata plants *plants have a large central vacuole, animals have much smaller vacuoles generally called vesicles

Cell Structures and Organelles in Eukaryotes

Cell walls Plant cells are surrounded by fairly rigid, nonliving walls composed of cellulose (a polymer of the sugar glucose) and other polysaccharides. The strength of the cell wall is based on the fact that the microfibril chains are arranged in crisscrossed layers. The result is a very strong but porous covering which can expand in young cells thanks to a certain elasticity and enzymatic rearrangement of microfibril components. As the cell wall ages hardening compounds such as lignin, (hemicellulose) and pectin impregnate the cellulose fibrils, preventing further growth. As plant cells divide a new cell wall is formed between them. The two new cell walls are cemented together with pectin to form a layer known as the middle lamella.

The cartoon above labels the parts of a plant cell visible under the light microscope.

A number of substances are deposited in or outside plant cell walls. For example:

lignin; technically an alcohol, thickens, hardens and prevents decay suberin and cutin; are waxy materials which acts as a water proof coating (to prevent excessive water loss.)

Modified July 8, 2005

Cytology - Part V
Plasma membrane (cell membrane) The plasma membrane is clearly not a passive envelope surrounding the cell, but is highly selective in what it allows to enter or exit. One of its essential functions is the maintenance ofhomeostasis. All cells need to regulate their internal environment by constantly removing waste products of metabolism and actively bringing in needed resources. The primary component of the plasma membrane is a double layer of phospholipids.

The bilayer mentioned above is common to all cellular membranes. (Taken from the WWW Cell Biology Course) What distinguishes one membrane from another is the proteins associated with or embedded in the phospholipids, which in turn is related to the membranes function.

Go to the University of Virginia for a professionally executed illustration of the cell membrane

Internal Support: The Cytoskeleton. What at one time was called the "cell matrix" or "ground substance" the seemingly clear regions of the cytoplasm have yielded their secrets with the use of fluorescent labeling, detergents and the high voltage transmission electron microscope. Today we know that much of the cytoplasm's matrix is laced with an intricate, mazelike network of proteinaceous microtubules intermediate filaments and microfilaments which constitutes the cytoskeleton. The cytoskeleton regulates cell shape, transport, motility, and integrity. It consists of 3 protein filament systems noted above and a large number of associated proteins that regulate each system. In summary the major functions of the cytoskeletal filaments and tubules are:

to act as a supporting framework for organelles to provide for cell movement to change or maintain cell shape to anchor the plasma membrane

The actin microfilaments and myosin motor microfilaments can be found wherever the cell membrane needs support and movement. This includes:

microvilli of intestine cells, cytoplasmic streaming, and transport or movement of organelles

Intermediate filaments of the epithelium are composed of the protein keratin, which lends strength to the microvilli as shown in the diagram below.

Microtubules are the largest of the cytoskeletal elements. A globular protein, called tubulin, is the building block of the microtubules, the tiny hollow tubes found throughout the cytosol. Each tubulin molecule is actually a dimer, composed of two spherical polypeptides. Tubulin dimers are assembled into spiraling rows quickly forming each microtubule. This modular structure allow the cell to rapidly convert the associated cytoplasm between a sol (solid) or a gel (fluid) state. _________________________________________________________________ Microtubules play a role in:

cytoskeletal structure intracellular material transport maintenance of cell shape cellular organization cell division and movement of chromosomes.

Specific cellular structures which are composed of microtubules are:

centrioles basal bodies cilia and flagella (click here for additional information) mitotic spindle apparatus (click here for diagram)

In cell division microtubules cross the cell from pole to pole, taking on a spindle shape. Each chromosome has numerous microtubules attached to its centromere (actually the kinetochore). Thus microtubules help to separate and move the daughter chromatids to the opposite poles of the cell.

The shape of specialized cells such as nerve cells will revert to a simple sphere when their microtubules are chemically disrupted. Learn more and about the cytoskeleton and practice some multiple choice questions at the Biology Project at Arizona State. Modified July 10, 2005

Cytology - Part VI
Control and Cell Reproduction: The Nucleus. (Visit the University of Texas - Medical Branch for a throughly illustrated lecture on the nucleus) When stained the nucleus is the most salient feature of a cell. This prominent organelle has dual functions:

metabolic control of cellular activities and cell reproduction (mitosis or meiosis)

Most of the cells DNA resides in the nucleus. The DNA combines with a variety of proteins to form linear molecules called chromosomes The chromosome may be functionally divided into a number of hereditary units called genes. Chromosomes, literally color bodies, are so named because they are easily stained by a number of pigments. A diagram of four nucleotide pairs of DNA's double molecular structure is shown below.

Within the nucleus are the nucleoli darkly stained bodies involved in intense RNA synthesis. This is where ribosomes are formed in a eukaryotic cell. When a cell is ready to divide chromosomes become visible using a light microscope because they thicken due to a highly coiled state. The nucleus has a double membrane surrounding it called the nuclear envelope. Scattered generously throughout the nuclear membrane are openings called nuclear pores which allow materials to pass back and forth between the nucleus and the cytoplasm. The nuclear pores are a complex of 8 adjoining proteins which form a circular cluster often seen with a central plug. Take a look at a freeze-fracture preparation of nuclear pores.

Click on individual labels in the diagram for electron micrographs and further explanation

Modified July 9, 2005

Cytology - Part VII

Organelles of Synthesis, Storage, and Cytoplasmic Transport

Endoplasmic Reticulum A complex and extensive system of convoluted membrane(s) taking up a sizable portion of the cytoplasm is known as the endoplasmic reticulum. The ER is a dynamic ever-changing structure; a continuous membrane which forms a closed sac. Its internal space or lumen, may account for 10% of cytoplasmic volume. The ER forms the outer membrane of the nuclear envelope so actually the nuclear material is separated from the ER lumen by a single membrane. The endoplasmic reticulum appears in two main forms smooth and rough. Note: In the cartoon of rough endoplasmic reticulum below many of the ribosomes (illustrated as dark spots) are much too regularly spaced.

The rough endoplasmic reticulum receives its name from the appearance of ribosomes, dense granules that stick temporarily to the outer sides of the endoplasmic reticulum. RER (rough endoplasmic reticulum) occurs in cells that are actively making proteins destined for secretion outside the cell -- examples of such proteins are antibodies, digestive enzymes, and certain hormones. The diagram below outlines the major steps in this process.

Ribosomes Ribosomes (2) are sites of protein synthesis. They are much too large and complex to be considered simple molecules.

A ribosome is actually made of two subunits each composed of rRNA (r = ribosomal) and many kinds of proteins which come together during polypeptide synthesis. The ribosomal subunits in bacteria are smaller and differ somewhat chemically from those of eukaryotes. It is this difference that makes many antibiotics at least temperarily effective against bacterial infection. Polyribosomes (or polysomes) are groups of free ribosomes held together like beads on a string by a molecule of RNA during protein synthesis.

Smooth endoplasmic reticulum The smooth ER is an intricate network of tubes and sacs found in most cells but is abundant in those cells involved in the synthesis, secretion, and storage of non-protein products such as phospholipids, cholesterol, and other lipids. Cells involved in lipid metabolism contain lots of SER. For example, steroid hormones are produced in the endocrine glands of the testis, and oils are produced by exocrine glands found in skin. It may be that SER in small intestine cells plays an important role in the way animals absorb and store the products of lipid digestion. The smooth ER in the liver has two functions:

it contains oxidizing enzymes that detoxify certain chemicals it is active in breaking down glycogen to glucose

Golgi apparatus Modern electron microscope studies of the Golgi complex show it to be a series of compressed baglike sacs or cisternae, generally found near the nucleus, and always associated with endoplasmic reticulum.

A stack of Golgi bodies in plants may be referred to as a dictyostome. The Golgi complex is one of the cells more dynamic organelles, constantly changing and busy. Golgi cisternae (protein filled sacs) actually originate from portions of the nearby endoplasmic reticulum known as cis vesicles The dynamics of the Golgi complex involves:

vesicles originating from nearby endoplasmic reticulum merge with the cis face of the Golgi apparatus the cisterna buds off vesicles at either end which then migrate to the next cisterna in the stack. eventually vesicles containing the finished cellular product bud from the trans face of the Golgi apparatus, and migrate through the cytoplasm to their final destination (usually the plasma membrane for exocytosis.)

A cartoon of the Golgi complex illustrates the dynamics described above.

Lysosomes Spherical bags of hydrolytic enzymes, synthesized in the RER and which mature in the Golgi complex are called lysosomes. The lysosome is able to make its own acidic condition by actively pumping H+ ions from the cytoplasm to its interior. The 40 or so hydrolytic enzymes so far discovered in lysosomes are most effective in an acidic environment. This is an important safeguard for most cells which have lysosomes otherwise the cell containing them would be a risk of self destruction.

Christian de Duve, the cell biologist who predicted the existence of lysosomes from chemical evidence, eventually found what he was looking for by using the electron microscope. Some lysosomes act as "suicide bags" because they are known to engage in autophagy. Apoptosis or programmed cell death, is now recognized as an essential part of multicellular development. Lysosomes are optically dense bodies -- they form continuously from the trans face of the Golgi complex, emerging as primary lysosomes. Visit the University of Virginia to view a detailed cartoon of lysosome formation and function. Three functions of lysosomes are: 1. digestion of large food particles (such as invading bacteria) 2. modification and storage of molecular materials obtained through endocytosis 3. destruction of aging or damaged cellular organelles Visit this following site for more information about lysosomes and other microsomes.

Microbodies Minor organelles, primarily sacs used to store or process various molecules include peroxisomes, glyoxysomes, and various vacuoles. a. peroxisomes are found in most eukaryotes. They probably have a protective function and may have been another organelle derived from the endosymbiosis with an ancient free living prokaryote. peroxisomes found in the liver contain the enzyme catalase which neutralizes dangerous peroxides such as H2O2. b. caveolae or "little caves" play a significant role in intercellular communication.

c. glyoxysomes are microbodies found in seeds and posses enzymes capable of mobilizing lipids, converting them into sugars to supply energy needed during germination. d. vacuoles are general all purpose storage containers. The large central vacuole of plants stores a watery solution which also regulates turgor pressure, the force plants use to stay turgid and enlarge developing cells.

Endomembrane System The endomembrane system is a network of shared and transfered membranes including the following components:

the outer membrane of the nuclear envelope smooth and rough endoplasmic reticulum Golgi apparatus and associated vesicles and the plasma membrane.

The endomembranal system is necessary to keep the products and reactants of metabolism separate and compartmentalized for appropriate disposal and to satisfy energy requirements. Make sure you observe the animated diagram at the bottom of this page

Modified July 9, 2005

Cytology - Part VIII

Energy Generating Organelles

Chloroplasts

Chloroplasts are found in plants and protista such as algae. Chloroplasts are not found in blue-green algae (cyanobacteria - kingdom Monera). However the cyanobacteria have their own membranes which function like the thylakoid membranes found in chloroplasts. The question of how plastids originated is being studied in a number of labs. The chloroplast is a relatively large complex organelle with a double membrane. These membranes create two compartments the stroma, site of the light indpendent reacions of photosynthesis (Calvin cycle) and the inner thylakoid membrane with processes electrons used by the light reactions. Chloroplasts have their own DNA (genes) and ribosomes. They are able to reproduce themselves and to carry out the synthesis of some (but not all) of their own proteins. The chloroplast reproduces by simply pinching in two which looks like fission in prokaryotes. Chloroplast means green form; green because the chloroplast contains the energy absorbing molecule chlorophyll. Chloroplasts are essential in the process of photosynthesis in which carbon dioxide and water are changed into sugar and other important organic molecules. Oxygen is released as a by-product. The cyanobacteria also contain energy gathering molecules related to the chlorophyll pigment found in eukaryotes. Inside its double membrane, each chloroplast contains a clear, watery area called the stroma, which corresponds to the cytoplasm of a cell. It is in the stroma where the chloroplast makes sugar in a metabolism called the Calvin cycle. This clear fluid area of the chloroplast is invaded by an extensive membrane system that forms saclike vesicles called thylakoids. The cartoon of a thylakoid below gives a diagramatic rendering of the major molecular components necessary for the light reaction of photosynthesis. A 3D view of one of the molecules involved in the harvesting of light in photosystem II may be viewed at the Lund University, Sweden

When thylakoid membranes are folded into stacks they are referred to as grana, and where these membranes occur singly they are called stroma thylakoids. The labeled illustration of a chloroplast found below may help you sort out the relationship between these terms.

Located in and on the thylakoid membranes are numerous granules called CF0CF1 complexes. The CF0 portion anchors the assemblage into the thylakoid membrane and forms a channel between the stroma and lumen, the two compartments of the chloroplast. The CF1 portion contains the enzyme ATP synthase, and as you might expect is involved in the synthesis of ATP. Besides the chloroplast, plants have other organelles called plastids which are related in origin but which have other functions.These include: a. leucoplasts or amyloplasts serve as storage for starch. They are often found in underground stems or roots. b. chromoplasts concentrate various pigments such as the carotenes and xanthophylls and are frequently found in the petals of flowers.

Visit the Virtual Cell to zoom into the thylakoid membranes and see how the light reaction works

Comparison of mitochondria and chloroplasts Three structures mitochondria and chloroplasts have in common are: 1. a double membrane 2. ribosomes capable of synthesizing proteins unique to the organelle 3. a single circular DNA molecule In addition both mitochondria and chloroplasts reproduce independently of the "host" cell. Mitochondria differ from chloroplasts because the final products of the mitochondrion are the reactants of the chloroplast. The use of sugar in aerobic respiration to produce energy by mitochondria is reversed in the chloroplast where energy is used to make sugar. Respiration is an exergonic reaction while photosynthesis is endergonic Many scientists propose that mitochondria and chloroplast are endosymbionts, that is these organelles are actually descendants of once-independent prokaryotic cells. The similarities between both mitochondria and chloroplasts and the prokaryotes is powerful evidence for the serial endosymbiotic theory for the origin of these organelles.

Mitochondria Mitochondria are generally much smaller than chloroplasts, at least in cross section. However mitochondria may have a surprisingly convoluted structure. In electron micrographs, mitochondria usually are oval, with the inner membrane exhibiting numerous folds called cristae. The apparent purpose for this extensive folding is to increase the surface area where most of the biochemical work of the mitochondria is accomplished. The following diagram illustrates relationships among the membranes and compartments found in mitochondria.

Electron microscope studies reveal that the cristae are covered with small, round bodies very similar to those in chloroplasts The spherical granules and their membrane bound tails are called F0F1 complexes which are similar to the CF0 CF1 complexes of chloroplasts. You can go to an animation of ATP synthase in action and visit a tutorial on metabolism. Cells rich in mitochondria have either high metabolic rates, or a need for explosive energy output such as skeletal muscles. For a complete discussion of mitochondrial structure and function visit the University of Texas Medical Branch site. Click here to visit other sites describing how mitochondria work.

Modified July 9, 2005

Cytology - Part IX
Organelles of Movement.

centrioles A centriole is an organelle found only in eukaryotes that develop cilia or flagella. They have two functions namely: a. form basal bodies which then give rise to cilia or flagella

b. somehow determine where the furrow forms in dividing cells

Centrioles are composed of two short cylindrical bundles of microtubules. Generally one centriole is at right angles to the other. Each centriole is a set of nine groups of microtubule pairs arranged in a pinwheel (see below).

Cilia and flagella are fine, hairlike, movable organelles found on the surfaces of some cells. Although these organelles appear to be outside the cell they are both surrounded by the plasma membrane. Cilia can be used to move a single cell as in the protist paramecium or they can be used to move material past a cell as in the epithelial cells lining the bronchial passages.

Links to Reading

Filming a Cellular Motor

Take a quiz on basic cell structure at the Biology 101 site.

Modified July 10, 2005