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LECTURE PRESENTATIONS For CAMPBELL BIOLOGY, NINTH EDITION Jane B. Reece, Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Robert B. Jackson Chapter 6 A Tour of the Cell Lectures by Erin Barley Kathleen Fitzpatrick © 2011 Pearson Education, Inc. Cells: The Fundamental Units of Life • first cell observed – Robert Hooke in 1665 • • thin slices of bottle cork saw a network of “cells” that a monk would live in • first living cell (Spirogyra) observed by Anton van Leeuwenhoek in 1674 • also described bacteria he found in his mouth (1676) Cells: The Fundamental Units of Life • the cell is the simplest collection of matter that can be alive • the Cell Theory: • 1. all organisms are made of at least one type of cell • 2. all cells come from pre-existing cells by this cell dividing in two • 3. the cell is the basic, fundamental unit of life • attributed to: Theodor Schwann, Matthias Jakob Schleiden, and Rudolf Virchow. Anton van Leeuwenhoek Cells: The Fundamental Units of Life • modern interpretation added a few more parameters – 1. The activity of an organism depends on the total activity of independent cells. – 2. Energy flow (metabolism and biochemistry) occurs within cells. – 3. Cells contain hereditary information (DNA) which is passed from cell to cell during cell division. – 4. All cells are basically the same in chemical composition in organisms of similar species. Concept: Biologists use microscopes and the tools of biochemistry to study cells • first compound microscope – Zacharias Jansen in 1590 • three important parameters of microscopy – Magnification: the ratio of an object’s image size to its real size – Resolution: the measure of the clarity of the image, or the minimum distance of two distinguishable points • inversely related to the wavelength of the radiation a microscope uses – Contrast: visible differences in parts of the sample Human height 1m • – • • Chicken egg 1 cm Frog egg 1 mm 100 m in compound microscopes – more than one lens – – • e.g. magnifying lens 0.1 m ocular and objective lenses improves resolution and allows for more than one magnification LMs can magnify effectively to about 1,000 times the size of the actual specimen this allows for individual cells within a tissue to be visualized Human egg Most plant and animal cells 10 m 1 m 100 nm Nucleus Most bacteria Mitochondrion Smallest bacteria Viruses Ribosomes 10 nm Proteins Lipids 1 nm 0.1 nm Small molecules Atoms Superresolution microscopy Electron microscopy • in a light microscope (LM) - visible light is passed through a specimen and then through glass lenses the lenses refract (bend) the light - so that the image is magnified in a simple microscope - there is one lens for magnification Light microscopy • Length of some nerve and muscle cells Unaided eye Studying Cells: Microscopy 10 m Light Microscopy (LM) Brightfield (unstained specimen) 50 m 50 m Brightfield (stained specimen) Deconvolution 10 m Phase-contrast Differential-interferencecontrast (Nomarski) Super-resolution Fluorescence 10 m 1 m • various techniques enhance contrast of a LM and enable cell components to be stained or labeled • BUT - most subcellular structures, including organelles, are too small to be resolved by an LM • LMs cannot resolve detail finer than 0.2um - regardless of magnification Confocal • • • • to improve resolution and magnification – allowing for imaging of subcellular structure development of two other microscopes in the 1950s called electron microscopes (EMs)- use a focused beam of electrons rather than light resolution increase – due to the shorter wavelength of the electron beam two types: – – 1. Transmission Electron Microscope (TEM) 2. Scanning Electron Microscope (SEM) • • two types: 1. Scanning electron microscopes (SEMs) – electron beam is focused onto the surface of a subject – – – providing images that look 3D SEM electron beam excites the electrons of the gold on the subject’s surface several kinds of electrons are produced • – – – Blood cells e.g. secondary and back-scattered these electrons are detected and projected onto a video screen as a magnified image that appears 3D can be colorized other detectors can be used • e.g. to detect elemental composition Pollen grains • 2. Transmission electron microscopes (TEMs) focus a beam of electrons through a specimen – – – – – – subject is sectioned first into a very thin layer – with a microtome TEMs are used mainly to study the internal structure of cells subject is stained with heavy metals that adhere to the internal structures of the cell so some parts of the cell are more electron dense than others the electron beam passes through those less dense and scattered by the more dense regions the electrons that pass through hit a piece of film negative or hit a detector for displaying the image Studying Cells: Cell Fractionation TECHNIQUE • another way to study cells is to study their individual components • subcellular fractionation takes cells apart and separates the major organelles from one another • centrifuges fractionate cells into their component parts – separates based on the density of the organelle within a separating solution – like sugar • cell fractionation enables scientists to determine the functions of organelles by being able to isolate them Homogenization Tissue cells Homogenate Centrifuged at 1,000 g (1,000 times the force of gravity) for 10 min Supernatant poured into next tube 20,000 g 20 min Pellet rich in nuclei and cellular debris Centrifugation Differential centrifugation 80,000 g 60 min 150,000 g 3 hr Pellet rich in mitochondria (and chloroplasts if cells are from a plant) Pellet rich in “microsomes” (pieces of plasma membranes and cells’ internal Pellet rich in membranes) ribosomes Concept: Eukaryotic cells have internal membranes that compartmentalize their functions • the basic structural and functional unit of every organism is the cell • two types of cells: prokaryotic or eukaryotic • prokaryotic cells: domains Bacteria and Archaea consist of prokaryotic cells • protists, fungi, animals, and plants all consist of eukaryotic cells Comparing Prokaryotic and Eukaryotic Cells • Basic features of all cells – Plasma membrane – Semifluid substance called cytosol – DNA in the form of Chromosomes (carry genes) – Ribosomes (make proteins) • Prokaryotic cells: – no nucleus – DNA in an unbound region called the nucleoid – no membrane-bound organelles – a Cytoplasm bound by the plasma membrane • Eukaryotic cells – membrane – bound nucleus containing DNA (nuclear envelope) – membrane-bound organelles – a Cytoplasm in the region between the plasma membrane and nucleus The Plasma Membrane • the plasma membrane is a semipermeable barrier – allows sufficient passage of oxygen, nutrients, and waste to service the volume of every cell • the general structure of a biological membrane is a double layer of phospholipids – known as a phospholipid bilayer – membrane proteins embedded within it or found attached to one layer Outside of cell Inside of cell 0.1 m (a) TEM of a plasma membrane Carbohydrate side chains Hydrophilic region Hydrophobic region Hydrophilic region Phospholipid Proteins (b) Structure of the plasma membrane The Cytosol • also known as the intracellular fluid or ICF • made up of multiple levels of organization: – concentration gradients of ions and small molecules – larger complexes of enzymes for metabolic pathways – large complexes of proteins • e.g. proteasomes in eukaryotes – protein degradation • e.g. carboxysomes in prokaryotes – carbon fixation (Calvin cycle – synthesis of sugar from CO2) • prokaryotic cells – site of the cell’s metabolic reactions The Cytosol – Eukaryotic Cells • eukaryotic cells – part of the cytoplasm – about 55% of the cell’s volume – about 70-90% water PLUS • • • • • ions dissolved nutrients – e.g. glucose soluble and insoluble proteins waste products macromolecules and their components - amino acids, fatty acids • ATP • unique composition with respect to extracellular fluid (ECF) • -fluid found outside of a cell Cytosol •higher K+ •lower Na+ •higher concentration of dissolved and suspended proteins (enzymes, organelles) •lower concentration of carbohydrates (due to catabolism) •larger reserves of amino acids (anabolism) ECF •lower K+ •higher Na+ •lower concentration of dissolved and suspended proteins •higher concentration of carbohydrates •smaller reserves of amino acids DNA & Chromosomes • one type of organizational unit of DNA= chromosomes • in eukaryotes each chromosome is composed of a single DNA molecule associated with proteins (histones) • prokaryotic chromosome = genophore – found in the nucleoid region of the cell – multiple copies may exist – usually just one circular piece of double-stranded DNA • means there is no need for telomeres – length varies – several hundred thousand to a few million base pairs • still significantly smaller than a eukaryotic chromosome – not really a chromosome – no chromatin • not found associated with histones proteins • can associated with nucleoid-associated proteins (NAPs) – play a role in spatially organizing the genophore within the nucleoid region • DNA is looped and supercoiled rather than compacted by histones Extrachromosomal DNA - Plasmids • • • • pieces of DNA not part of the genophore/chromosome replicate independently of the genophore substantially smaller – usually only a few thousand base pairs can be found in all three domains: Bacteria, Archaea and Eukaryota • carry genes that confer antibiotic resistance to prokaryotes • used in the laboratory for manipulation of genes The Eukaryotic Nucleus: Information Central • the nucleus contains most of the eukaryotic cell’s genes and is usually the most conspicuous organelle • the nuclear envelope encloses the nucleus, separating it from the cytoplasm – not found in prokaryotic cells – so NO nucleus in prokaryotic cells • the nuclear envelope is comprised of two phospholipid bilayers – each membrane is a lipid bilayer – often connected to the endoplasmic reticulum • the nucleolus is located within the nucleus and is the site of ribosomal RNA (rRNA) synthesis – more synthetic the cell is – the greater number of nucleoli Nucleus Nucleolus Chromatin Nuclear envelope: Inner membrane Outer membrane Nuclear pore Rough ER Pore complex Ribosome Close-up of nuclear envelope Chromatin Ribosomes: Protein Factories • Ribosomes are particles made of ribosomal RNA (rRNA) and protein • comprised of two subunits – large – small • ribosomes carry out protein synthesis in two locations – in the cytosol (free ribosomes) – both prokaryotes & eukaryotes – in eukaryotes - on the outside of the endoplasmic reticulum or the nuclear envelope (bound ribosomes) • can also be found in chloroplasts and mitochondria 0.25 m Free ribosomes in cytosol Endoplasmic reticulum (ER) Ribosomes bound to ER Large subunit Small subunit TEM showing ER and ribosomes Diagram of a ribosome Prokaryotic Cells • usually smaller and less complex than eukaryotic cells • first cells to evolve • single-celled organisms – 0.5 to 5um in diameter (eukaryotes 10-100um) – some species exist as aggregates called colonies • e.g. cyanobacteria • some prokaryotes can be multi-cellular at points in their life-cycle • defined as having no nuclear membrane, no mitochondria or other organelles • divided into two domains: – Bacteria – Archaea Prokaryotic Cells • Bacteria: – collective biomass – 10x of all eukaryotes – vast genetic diversity among members – physical diversity • shapes: spheres (coccus), rods (bacilli) and spirals 1 µm Spherical (cocci) 2 µm Rod-shaped (bacilli) 5 µm Spiral Prokaryotic Cells Fimbriae Nucleoid • made up of numerous components: – cell capsule – cell wall – plasma membrane – fimbrae – flagella(e) – cytoplasm – nucleoid region – ribosomes Ribosomes Plasma membrane Bacterial chromosome Cell wall Capsule Flagella (a) A typical rod-shaped bacterium cell wall capsule plasma membrane genophore cytoplasmic granules plasmid poly-ribosomes ribosome subunits Prokaryotic Cells Fimbriae Nucleoid • Bacterial cell components: – nucleoid region – nonmembranous region where the DNA is located • • • • 60% DNA 40% protein and RNA (mRNA) DNA is a circular chromosome other smaller pieces of circular DNA are located here – plasmids (house genes for antibiotic resistance) – ribosomes - for protein synthesis Ribosomes Plasma membrane Bacterial chromosome Cell wall Capsule (a) A typical rod-shaped bacterium Flagella Prokaryotic Cells Fimbriae Nucleoid • Bacterial cell components: – plasma membrane – phospholipid bilayer similar to that of eukaryotes – cell wall – rigid structure outside the PM • composed of a peptidoglycan layer (crosslinked sugars) • for protection, structural support & reproduction • gram negative bacteria have an additional layer outside the cell wall – called a lipopolysaccharide layer • this layer is responsible for the toxicity of these bacteria – capsule - also called the glycocalyx or “slime layer” • for adherence Ribosomes Plasma membrane Bacterial chromosome Cell wall Capsule (a) A typical rod-shaped bacterium Flagella Prokaryotic Cells Fimbriae Nucleoid • Bacterial cell components: – fimbrae – structures involved in attachment • shorter ones involved in reproduction = pilus Ribosomes Plasma membrane Bacterial chromosome – flagella – made of protein filaments • for locomotion – cytoplasm – intracellular fluid of the prokaryote • bound by the plasma membrane • no membrane-bound organelles Cell wall Capsule (a) A typical rod-shaped bacterium Flagella The Eukaryotic Cell • cells of the Domain Eukaryota • a eukaryotic cell has internal membranes that partition the cell into organelles – known as the endomembrane system • single eukaryotic cells found in the Kingdom Protista • remaining Phyla in the domain contain mainly multicellular organisms – fungus, plants and animals • plant and animal cells have most of the same organelles The Eukaryotic cell • 1. Plasma membrane • 2. Nucleus • 3. Cytoplasm – a. cytosol – b. cytoskeleton • 4. Cytoplasmic organelles – endomembrane system Outside of the plasma membrane will be a Cell Wall in plants and fungus – not the same composition