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Intro Microanatomy

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1) Name the components that make up cells, tissues and organs. Cells are comprised of three domains:  Nucleus  Intracisternal (inside organelles)  Cytosol (Cytoplasm contains organelles  Intracisternal space) o Contains cytoskeletal elements Tissues: Cells + ECM (Extracellular Matrix)  4 Types of Tissue o Epithelial: closely linked cells form a lining o Connective Tissue: Supportive tissue can be flexible or rigid (cartilage vs bone) o Nervous Tissue: Specialized for conduction o Contractile Tissue: Muscles with contractile protein fibers Organs: Comprised of Multiple Tissue Types (Usually all 4)  Complex structure Organ Systems 2) List common features of cells - Domains (i.e Organelles) have specific functions and are Separated by Membranes Know: (1) nucleolus (2) nucleus (3) ribosomes (4) vesicle (5) rough endoplasmic reticulum (ER) (6) Golgi apparatus (7) cytoskeleton (8) smooth ER (9) mitochondria (10) vacuole (11) cytosol - Different from Inclusions which are substances stored in cell (Glycogen, Fat)

Membranes: Phospholipid Bi-layer establishes compartmentalization / domains  Restricts / Regulates movement in and out of cell  Fluid-Mosaic Model: Membrane has lipid and protein components o Small polar or charged molecules move via Protein Mediated Transport o Large Molecules move through vacuoles through Endocytosis and Exocytosis o Integral Membrane Proteins: contain hydrophobic domains are require detergent to remove from membrane o Peripheral Membrane Proteins: Do not completely penetrate bi-layer and are only temporarily associated with membrane

Glycoconjugates: Sugars associated with membrane proteins o Cell-cell interactions  Recognition o Cell-matrix interactions o Almost Exclusively on Non-Cytosolic Side

Polarity: Non-Random distribution of organelles  Tight-Junctions cause polarity in Membrane proteins 3) Explain the basic processes of tissue preparation and microscopy for histological preparations 1) Fixation: Cross linking of proteins with formaldehyde. Need stronger fixatives like glutaraldehyde preserves ultrastructure 2) Sectioning: Fixed sample is sliced so light can pass through it on a slide     < 50 microns thick for light microscopy (hair is 100 microns) Electron microscopy needs 60 nanometers o Black and White Use Microtome Plane of Section

3) Staining  Hematoxylin: Stains basophilic subtances blue o DNA, Rough ER  Eosin: Stains acidophilic substances pink o Mitochondria, Cytoplasm

Protein Structure and Function

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Prior Review 1. pH of strong and weak acids Stronger acids have a lower pH than weak acids. Weak acids tend to have better buffering properties, especially when mixed with weak bases. 2. pK, buffers (Henderson-Hasselbach), titration curve pKa is the acid dissociation constant, pKb is the base dissociation constant. Stronger acids have a larger pKa. As stolen from wikipedia, if an acid dissociates like HA

Then the

The Henderson-Hasselbalch Equation states that

Namely, the pH of a solution depends on the extent of dissociation of its dissolved acid (a similar equation exists for basic solutions). A buffer solution resists change in pH from the addition of acids or bases up to a certain point. It‟s usually a mixture of a weak acid and weak base. As more acid is added to the buffer, the weak base dissociates to raise the pH. When all of the base has dissociated, the buffer “breaks”. The reverse is true, of course, when the weak acid dissociates to resist an increase in pH. 3. General Properties of Amino Acids An amino acid has a carboxyl (COO-) end, an amino (NH2-) end, and a side chain. Each side chain has its own pKa. Depending on the pH, this will give the chain a charge. The isoelectric point is the pH at which a specific side chain will have no charge. In general, amino acids are assigned a charge

based on standard body pH, although there are environments in the body with extremely low and high pH. 4. Structure of the peptide bond The peptide bond links together amino acids from carboxyl end to amino end via dehydration synthesis. It‟s a covalent bond and difficult to break. Learning Objectives 1. General Properties of proteins Functions: Seen in table below. Catalysis Regulation Transportation Contractile elements Defense Structural elements Size: molecular weight ranges from 6000 to 40,000,000 Shape: Globular Fibrous Conjugated: Protein and Sugar/Lipid/RNA covalently linked Charge: Depends on amino acids on surface of protein. Solubilitiy: Depends on location in body. Blood proteins are water soluble, membrane proteins lipid soluble. Some proteins are amphipathic. 2. Levels of Protein Structure Primary: Amino acid chain derived directly from gene translation. Each unit is connected via peptide bonds  Determines higher orders of folding Secondary: Smallest possible structure, connection by weak hydrogen bonds. Most common structures are the alpha helix and beta pleated sheet.  H-bonds  localized

Tertiary: Three-dimensional structure of a single protein chain. Stabilized by hydrogen and ionic bonds. Folding usually needs to be done exactly right in order for protein to function properly. Chaperonin proteins exist to refold damaged proteins correctly, usually by presenting them with a rapidly alternating hydrophobic and hydrophilic environment.  Ribonucleases always fold correctly after denaturation  insulin is irreparable when denatured. Quaternary: The conjugation of multiple tertiary protein structures. Stabilization by hydrogen, ionic, and hydrophobic interactions. For example, hemoglobin, collagen. 3. Protein folding and denaturation Discussed above. 4. Structure-function relationships Protein structure is related to function. Globular proteins usually have hydrophobic center and hydrophilic surface. Enzymes have a unique active site that binds to a specific substrate. Examples given in class: A point mutation on a B-chain of hemoglobin results in a long chain instead of a globular protein. The protein loses its solubility and sickle cell results. A mutation in CFTR makes it unable to transport Cl- and cystic fibrosis results.

Enzymes

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1. General properties of enzymes: -Enzymes are a class of proteins that increase the rate of a chemical reaction. -Because enzymes control the rates of reactions, they are used to regulate the activity of the cell. -Enzymes have a specific distribution within subcellular compartments and within specific organs. *As proteins, enzymes are sensitive to changes in temperature and pH and require a relatively stable environment in order to function. *Enzymes are often kept in the inactive state, where it is called the zymogen or proenzyme. This allows enzyme activity to be strictly regulated. Example: Many proenzymes require s short sequence on the N-terminus to be cleaved in order to become active. For example, pepsinogen is translated and released by chief cells in the stomach. Trypsin then cleaves the Nterminus, converting the proenzyme to its active form pepsin. 2. Interaction of enzyme with substrate: -Substrates bind to a relatively small region of an enzyme called the active site. The bound substrate fits in a specific orientation and is fitted through ionic bonds, hydrogen bonds, and hydrophobic interactions. - No covalent boding to enzyme -The act of the substrate binding to the enzyme can cause a conformational change in the enzyme. This is also called induced fit. - Enzymes don‟t affect the Thermodynamics, they do effect the kinetics 3. Enzyme catalysis; Michaelis-Menten equation -Enzymes have no effect on the thermodynamic properties of a given reaction and therefore always move the chemical reaction towards equilibrium. Instead, enzymes lower the energy of activation, an energy barrier required in order for a reaction to proceed, and thereby increase the speed of a reaction. -Enzymatic reactions can proceed in the forward or backward reactions depending on where the chemical equilibrium lies. -The Michaelis-Menten equation allows one to predict the rate of reaction given a specific amount of substrate.

*During catalysis, the enzyme remains unchanged after the reaction has taken place. In many cases, the enzyme forms a covalent intermediate. However, this covalent bond is not involved in substrate-enzyme binding. 4. Enzyme inhibition -Competitive inhibitors directly compete with the substrate to bind at the active site. -A competitive inhibitor will increase the Km, the concentration required for half the enzymes to be bound to substrate, because the competitive inhibitors will always occupy a specific portion of active enzymes. -A competitive inhibitor will leave vmax unchanged because adding an infinite amount of substrate will allow the enzyme to bind to the substrate more often than to the competitor.

-Noncompetitive inhibitors, also called allosteric inhibitors, bind to a site on the enzyme somewhere other than at the active site. -Non-competitive inhibitors will occupy a given portion of enzymes at any given time, thereby reducing vmax regardless of substrate concentration. *It is hypothesized that the noncompetitive inhibitor binds to the enzyme and prevents it from achieving a specific conformational state, thereby making the enzyme non-functional. 5. Mechanism of enzyme reactions

-Enzymes can have a high specificity to a given substrate or can be more non-specific (digestive enzymes). -In a given reaction with an enzyme, two reactants need to bump into each other with the proper orientation in order for the reaction to take place. An enzyme binds to these substrates, thereby increasing effective proximity and placing the substrates into the proper orientation. *An enzyme remains unchanged after performing the appropriate chemical reaction. 6. Regulation of enzyme activity -Isozymes are multiple forms of the same enzyme, often with different kinetic properties. ex/ -Lactate dehydrogenase is given as a specific example where the distribution of lactate dehydrogenase is specific to different organs. -Phosphorylation can activate or deactivate a given enzyme. -Positive and negative modulators can demonstrate cooperativity (speeding up reaction) or inhibition in order to alter the kinetics of the reaction.

*indicates relevant information covered in other lectures but not this one

DNA Replication
DNA REPLICATION 1. Phase Initiation

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Compare and contrast DNA replication in Prokaryotes and Eukaryotes Prokaryotes DNA A binds to OriC (only origin of replication) and melts DNA DNA B (helicase) unwinds DNA Topoisomerase I needed to nick 1 strand of DNA to relieve torsional stress (bc continuous circle) Single Strand Binding Proteins (SSBs) bind to prevent DNA from re-binding to other parent strand RPAs bind to prevent DNA from re-binding to parent strand Primase recruited to replication fork and adds RNA primer to leading strand, then to lagging strand further down DNA DNA Pol α adds a few DNA nucleotides to primer Eukaryotes

Helicase(?) binds to origin of replication (many)

Helicase (?) unwinds DNA

Priming

Primase recruited to replication fork and adds RNA primer to leading strand, then to lagging strand further down DNA

Elongation

DNA pol III then binds (tethers with beta clamp) to polymerize DNA in 5‟-3‟ direction (leading strand in

DNA pol δ then binds (tethers w PCNA) to replicate in 5'-3' (leading strand in continuous manner, and

continuous manner, and lagging strand in discontinuous manner w Okazaki fragments) DNA Pol III can backtrack and proofread in 3‟-5‟ direction DNA Pol I replaces DNA Pol III to remove RNA primers

lagging strand in discontinuous manner w Okazaki fragments)

DNA pol δ can backtrack and proofread in 3'-5' direction Fen-1 removes primers and DNA pol δ replaces gaps w DNA DNA ligase joins strands Telomerase creates RNA template to extend lagging strand with junk DNA

DNA ligase joins strands Termination Terminator sequences trap replication fork near origin site and bing TUS proteins 2nd TUS Protein does not allow DNA B to pass through, and elongation is stopped Topoisomerase iv unlinks the catenated strands

T-loops formed at ends

2.

Examples of diseases that occur due to replication defects

a. Mutation in RNA telomerase --> Dyskeratosis congenity: Developmental delay b. Low telomerase levels --> no T-loops = genomic instability = increased cancer risk c. Fragile X syndrome – excess CGG d. Muscular dystrophy e. Spinocerebellar ataxia f. Huntington‟s Disease: In coding sequence

Mutations in UTR, introns and coding sequence

DNA Mutation and Repair

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1. Describe the relationship between DNA damage, DNA repair, DNA replication and mutagenesis Mutagenesis: the permanent alteration of DNA

2. State the major sources of DNA damage and the major types of DNA repair Sources of DNA Damage 1. Endogenous Sources DNA Replication Errors (misincorporation, slippage) Deamination (cytosine to uracil, 5-methylcytosine to thymine) Depurination (creates abasic site) Reactive Oxygen Species (strand breaks, base damage) DNA Recombination Errors 2. Environmental Sources: -Ionizing Radiation (IR) increases reactive species (Indirect Mechanism) -Ultraviolet Radiation (UV) generates pyrimidine dimers (Direct Mechanism) -Chemical Mutagens (e.g. alkylation by MNNG, MNU → O6-methylguanine, pairs like A) Repair – least to most serious Proofreading – during replication. error rate:10^-4 10^-8 Post Replication Mismatch excision repair– after replication. Error rate 10^-8 10^10 Error recognition strand discrimination->excision->resynthesis->ligation Base excision repair: DNA glycosylase flips and removes base AP endonuclease cuts phosphodiester bond -> DNA polymerase -> ligase - can create abasic site that can be premutagenic if not repaired on time Direct reversal: MGMT destroys itself to get rid of methylation of guanine bases

Nucleotide excision repair: Damage recognition Nuclease cleavage removal with helicase Polδ, Polε DNA ligase removes bulky dimmers/unrecognizable bases/ note: DNA ligase repairs single-strand breaks > 100,000 / day  very efficient Homologous Recombination – less errors, only available during mitosis when sister chromatid is around. Reliable repair of double strand breaks exonuclease cuts to make stick endstrand invasion by sister chromatidDNA synthesis/sister chromatid exchangeunwindig/ligation(BLM helicase) Non-homologous Recombination – More error prone, available any time of cell cycle. Unreliable – possible error in relegation, clean up step is wasteful synapse formation to hold ends together by Ku70 and Ku80 DNA PKs clean up staggered ends Ligation by LIG4 and XRCC4 proteins

3. Describe the clinical consequences of mutagenesis and of defects in DNA repair

Nuclear Structure and Function

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1. Make a simplified schema of the basic structural components of the nucleus (nuclear envelope, pores, nucleolus, chromatin, nuclear matrix).

2. Explain the structure and function of the nuclear envelope and nuclear matrix.
Nuclear Envelope: Is a 2-membrane system which forms a barrier between the nucleus and the rest of the cell to: - Maintain unique protein and nucleotide environment - Sequester mRNA synthesis from Translational machinery - Protect / Contain DNA Outer Membrane and Inner Membrane are separated by the Perinuclear cisterna. It is perforated by Nuclear Pores containing NPC (Nuclear Pore Complexes). The inner membrane is supported by the Lamina

Nuclear Matrix: A protein lattice made of fibers (similar to cytoskeleton) which: - Anchors DNA replication - Anchors transcription complexes - Reinforces nuclear envelope --> stability

3. Explain the importance of the structure of nuclear pore complexes and how this plays a role in nucleocytoplasmic exchange. Nuclear Pore Complex: (80-100 nm diameter) is the site of selective nucleocytoplasmic exchange. It creates a selective barriers for the transport of macromolecules across the nuclear envelope (not as selective for smaller molecules). Has a 3-ring Structure that has 8 spokes with a hole in the center. - Cytoplasmic Ring: (8-subunits) with protusions into cytoplasm for mediating import - Middle Ring: 8 subsunits protrude into Perinuclear space o Transport proteins Nucleoplasmic Ring: Fibrous proteins protrude into nucleoplasm  Serves as a dock for importins and exportins. o Importin / exportin mediated transport requires energy

4. Describe the organization of chromatin and its role in synthesis, processing and storage of DNA and RNA. Chromatin is highly compacted DNA + Packing Proteins: - Heterochromatin: highly coiled and less active Genes not transcribed in cell - Euchromatin: less coiled, more active DNA Composition: Nucleosome: (10 nm) DNA wrapped around 8 Histones (166 bp/ histone)  Forms Beads on a String Coiled Nucleosome: (30 nm) coiled nucleosomes  H1 Histone: Activity affects density of nucleosome coil

The density of chromatin packing determines the availability of the genes for transcription. Histone tails are sites for activating the exposing / coiling of local DNA 5. Have a general concept of the cell cycle and its control as well as apoptosis. G1: Cell Growth S phase: DNA replication G2 phase: Interval before mitosis (valuable for DNA lesion repair) Mitosis

RNA Transcription

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Transcription and Control of Transcription Learning Objectives 1. Describe the basic transcription machinery, the basic structure of genes (including promoters) and transcription units, and the basic mechanism of transcription in eukaryotes. a. Basic machinery needed i. RNA pol (to read your template 3‟-5‟) ii. Some bases (ATP, GTP, CTP, UTP, all ribonucleotides of course) iii. DNA topoisomerases to unwind the helix b. Basic structure of genes, w/promoters & txpn units  ii. Promoter region o 1. Where proteins bind to begin transcription. This includes: o 2. Initiator sequence (which includes the…) o 3. TATA box o 4. A mix of enhancer and silencer sequences  a. Can be in other places other than right before the transcribed gene (ex. Behind, in the intron, etc.)  b. Fxn: assist regulation by allowing a specific txpn factor to bind to it c. This leads to activation/repression of transcription d. Environmental conditions can control the binding of txpn factors to these enhancer/silencer elements  e. Ultimately, the binding will lead to actions such as phosphorylation or binding/dissociation of another protein that is related to the txpn factor iii. Transcribed gene Exon: leaves the nucleus as mature mRNA afte modification  Intron: Kept inside the nucleus (although problems with the intron   could later contribute with mutations and problems with the mature mRNA) c. Basic mechanism of euk txpn - RNA pol transcribes the DNA - Depending on the RNA we are attempting to transcribe, we will use a corresponding polymerase - Basal txpn factors assist pol in recognizing the promoter and initiating txpn -NOTE: Mitochondria (have RNA pol that is similar to prok pol, and

transcribes their own DNA into their own rRNAs, mRNAs, and tRNAs)

2.Discuss the roles of transcriptional activator proteins, enhancer elements, coactivators, and chromatin in regulation of eukaryotic transcription a. Transcriptional activator proteins - Bind basal txpn factors associated with RNA pol 2 to get it over to the promoter - Recruit coactivators to perform 2 functions  Coactivators are proteins that increase gene expression by binding to an activator or txpn factor which contains a DNA binding domain, facilitating the txpn of a desired gene  Alter chromatin structure (like unwind it from the histone) to make promoter region more accessible  Recruit RNA pol II and its basal transcription factors - Enhancer elements (gene sequences far upstream/downstream for the gene or nearby) are brought closer to the gene we want to transcribe through complexes of transcriptional activator proteins, coactivators, and other transcription factor proteins in preparation for transcription by RNA pol II

3. Describe the cellular response (or signal transduction) pathway used by steroid hormones and list the major hormones which interact with members of the nuclear rece with the steroid receptor protein family also known as the „nuclear receptor‟ family  Glucocorticoids, Mineralcorticoids, Estrogens, Androgens, Progestin  Can also interact with steroid-related vitamins, amino acid derivatives, and other molecules yet to be discovered

b. Pathway - Steroid comes into the cell and is bound by a steroid receptor - This creates a steroid-protein complex that enters the nucleus, which binds to a hormone enhancer element on DNA  Steroid Receptor can be stabilized as dimmers after binding Steroid - The bound complex + enhancer sequence will fold up/join the promoter region, which will now begin to bind txpn factors, coactivators, and Pol II onto the promoter region. The TATA box is illustrated in the example above to give a frame of reference. - Now the desired gene can be transcribed into mRNA - The mRNA is then modified and packaged so it can exit the nucleus and be translated into protein - This protein will in turn create a physiological response

4. Explain why agnoists promote gene activation by steroid receptors, but antagonists inhibit steroid receptor function Agonist binding steps Antagonist binding steps

1. Agonist molecule binds to a receptor. In our notes, the receptor is a steroid receptor. 2.Receptor binds to enhancer sequence on DNA

1. Antagonist molecule binds to a receptor

2.Receptor binds to enhancer sequence on DNA

3.Receptor undergoes a conformational change, yielding a new binding spot

3.Receptor undergoes a conformational change, BUT there is NO new binding site

4.A coactivator protein will bind to this new spot, and with this binding, will recruit the binding of other transcriptional factors and RNA Pol 2 5.Now transcription can occur :)

4.Coactivator has no place to bind, the txpn apparatus never sets up

5.No transcription occurs :(

Conclusion: promotion of activity

Conclusion: inhibited acitivity

5.Discuss the roles of steroid receptors and their agonist/antagonists in the etiology and/or treatment of breast cancer  Breast tissue development is triggered by estrogen  Therefore, estrogen is an agonist for breast tissue/breast cancer growth

Tamoxifen, an anti-cancer drug, is an antagonist, changing conformation and inhibiting txpn by denying coactivators and txpn factors a binding site (see question 4) This prevents the growth of breast cancer cells

6. Explain how the cAMP signaling pathway can regulate txpn of specific genes a. Ex. Glucagon pathway (which signals that we need to make glucose) i. Protein or steroid from outside the cell binds to a receptor. ii. The receptor activates a G protein, which activates adenylyl cyclase iii. Adenylyl cyclase releases cAMP, which binds to protein kinase A iv. Protein kinase A enters the nucleus via nuclear pore, phosphorylating CREB (cAMP response element binding) protein. Now this is just like question 3! v. CREB now binds to its enhancer region, CRE (cAMP reponse element) vi. NOT in the notes, but I thought this was helpful: a coactivator called CBP (CREB-binding protein) then binds to CRE vii. Now txpn is activated

9/29/2011 10:01:00 AM ---------------------------------------Regulation of Transcription------------------------------------Initiation Can have multiple promoter and start sites Creates diversity by including/excluding exons Also changes the UTR length and potential for regulation Capping: Guanine cap Capped by inverted guanine Some groups are methylated Done by capping enzymes associated with polymerase as it transcribes

-

- recognized by nuclear pores, necessary for proper export - prevents exonuclease degradation - promotes circularization and translation Polyadenylation - Transcription ends when it recognizes termination sequence AAUAAA - Also can have multiple termination sites  provides 3‟ end diversity Once termination sequence is recognized, mRNA is cut 30 bp down and 200 adenosines are added - Necessary for export of the mRNA --> proteins bind poly A tail  Link to cap to help promote translation  Prevents 3‟ end exonuclease degradation  Splicing Introns out, Exons in Alternative splicing creates great biodiversity (when intentional) Temporal/spatial regulation Consensus site is strong for introns Excised structure is termed „lariat‟ When accidental or mis-spliced, can be harmful to cell  Dominant negative forms  Protein complexes (snRNPs) remain on mRNA, cells can tell if intron is left in --> Tend not to be exported Cryptic sites- sites (sometimes mutations) that become splice acceptor/donor sites that are not the normal sites Ex/ Portuguese family with cystic fibrosis cryptic splice site >> frameshift mutation - Burkitt‟s Lymphoma - chromosomal translocation shortens 3‟ UTR, removing sequences necessary for mRNA downregulation

RNA Translation

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1) Describe the principle of mRNA translation and explain the degeneracy of genetic code a. mRNA is read in groups of three in two-unit ribosome complexes where tRNA match the mRNA and bring the appropriate amino acids. They form peptide bonds to form the primary structure of the protein b. Degeneracy: There are 64 possible combinations of 3 basepairs, and only 20 amino acids. Each amino acid is coded by 2-6 codons 2) Understand and be able to summarize the general steps of translation

a. Scanning and initiation (2 ATP for aa-tRNA synth; 1 GTP required to unwind 5‟ UTR) i. Aminoacyl-tRNA synthetases couple amino acids to the correct tRNA ii. 40S ribosomal subunit binds to 5‟ cap and starts scanning 5‟-3‟ iii. Finds AUG start codon (ACCAUGG = Kozak sequence – usually start) iv. tRNA recognizes codon & brings in Met v. 60S ribosomal subunit joins for protein synthesis b. Elongation (2 GTP / amino acid) i. AAs bind to A site, then P site as new peptide bonds form. (Leave via E site) 1. This is called Translocation ii. N terminus = 5‟, C = 3‟ c. Termination (1 GTP) i. Occurs when encounters stop codon ii. Requires Release Factor R + GTP iii. Polypeptide released iv. Likely a physical link bw cap and poly A tail --> ribosome recycling 3) a. Explain how aberrant translation can play a role in human: Splicing mutations/ frameshift changes i. Occurs if introns not removed or exons are skipped & not matched up correctly

ii. Frameshift will change all subsequent amino acids & will result in incorrect protein iii. Only a frameshift of 3 amino acids would be less problematic but may still cause a problem w the protein b. The role of nonsense-mediated mRNA decay i. A frameshift mutation will likely cause an early stop codon ii. If a stop codon is encountered, the ribosome releases a release factor that scans for exon/exon junction complexes (EJCs) 50+ bp upstream of stop codon iii. If a protein is encountered, the mRNA is signaled for destruction iv. This is important for cells because dominant negative proteins can be very detrimental to cells (eg beta thalassemia = shortened beta subunit of hemoglobin ruins all hemoglobin)

Protein Secretion and Trafficking

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1) Describe the general mechanism of protein synthesis, and where it takes place in epithelial cells. Protein synthesis takes place on ribosomes. The ribosome life cycle Ribosomal RNA synthesized in nucleolus Ribosomal proteins synthesized in cytoplasm Ribosomal proteins imported to nucleolus, two subunits assembled (The two eukaryotic ribosomal subunits are 40s and 60s, useful to remember. Also useful numbers for the CF sweat test: <40mosm is negative, >60mosm is positive.) Ribosomal subunits are transported to cytoplasm. Small subunits identify and bind to mRNA Large subunits join to complex to form the ribosome. (The two eukaryotic ribosomal subunits assemble to form an 80s protein. Minimum sweat needed for the sweat test is 75 mg.) FROM 2011 FIRST AID FOR THE USMLE STEP 1 Protein Synthesis Initiation Activated by GTP hydrolysis, initiation factors help assemble the 40s ribosomal subunit with the initiator tRNA and are released when the mRNA and the ribosomal subunit assemble with the complex. Elongation Aminoacyl-tRNA binds to A site (except for initiator methionine) Ribosomal rRNA (ribozyme) catalyzes peptide bond formation, transfers growing polypeptide to amino acid in A site. Ribosome advances 3 nucleotides toward 3‟ end of RNA, moving peptidyl RNA to P site (translocation) Termination Stop codon is recognized by release factor, and completed protein is released from ribosome. Mnemonics Eukaryotes: 40S+60S=80S (Even) prOkaryotes: 30S+50S=70S (Odd) ATP = tRNA Activation

GTP = tRNA Gripping and Going places (translocation) Clinical Relevance Many antibiotics act as protein synthesis inhibitors Aminoglycosides inhibit formation of the initiation complex and cause misreading of mRNA Chloramphenicol inhibits 50S peptidyltransferase Macrolides and clindamycin bind 50S, blocking translocation. Ribosome location Free ribosomes in cytoplasm vs. Bound ribosomes on ER. Both form polyribosomes (polysomes). 2) Diagram and list the basic functions of the RER, SER, and Golgi complex. Rough Endoplasmic Reticulum

FROM 2011 FIRST AID FOR THE USMLE STEP 1 Nissl Bodies (RER in neurons) synthesize enzymes (e.g. ChAT) and peptide neurotransmitters. Mucus-secreting goblet cells of small intestine and antibody-secreting plasma cells are rich in RER. Smooth Endoplasmic Reticulum

Irregular network of membrane-bound tubules. Functions in steroid hormone synthesis, drug detoxification, and release and recapture of calcium ions for muscle contraction. FROM 2011 FIRST AID FOR THE USMLE STEP 1 Liver hepatocytes and syeroid hormone-producing cells of the adrenal cortex are rich in SER. Golgi Complex

FROM 2011 FIRST AID FOR THE USMLE STEP 1 Golgi apparatus Distribution center of proteins and lipids from ER to plasma membrane, lysosomes, and secretory vesicles. Modifies N-oligosaccharides on asparagine. Adds O-oligosaccharides to serine and threonine residues

Addition of mannose-6-phosphate to specific lysosomal proteins --> targets the protein to the lysosome Proteoglycan assembly from core proteins Sufation of sugars in proteoglycans and of selected tyrosine on proteins. Vesicular trafficking proteins: COPI: Retrograde, Golgi --> ER and more  Constituitive secretion  Intercisternal transport COPII: Anterograde, RER -> Golgi Clathrin: trans-Golgi --> lysosomes, plasma membrane --> endosomes

3) Distinguish between regulated and constitutive secretory pathways. Constitutive Secretion (default pathway): Usually continuous, needs no special stimulus. Can be upregulated or downregulated. (cop 1) Regulated Secretion: Requires an extracellular stimulus. Until then, proteins to be secreted are concentrated into secretory granules.

Cytoskeleton
Actin Microfilaments Subunits Made from globular (g) actin Tubulin Microtubules Made from tubulin dimers (alpha and beta) Dimers align to form protofilaments.

9/29/2011 10:01:00 AM Intermediate Filaments Various elongated proteins, often helical Dimers and tetramers

Primary Structures

Form filamentous actin (f-actin) in a coiled microfilament

Higher order structures

F-actin MFs can run parallel or anti-parallel, branch, be capped, anchored, or severed (via protein partners) ~7 nm ATP

Protofilaments (13) assemble side-side to form hollow tubules

Coiled-coil (likely not important)

Size Use nucleotide? Polarity

~25 nm (hollow) GTP

~10 nm Nope.

assembly usually occurs on (+) end, disassembly on the (-) end

(-) is anchored centrally to the MTOC, or to the basal body. Assembly usually occurs on (+) end extending towards cell periphery

None

Dynamics Motors

Fast Myosin

Fast Kinesin (+) and dynein (-)

Very slow ?

Found in:

-Cell peripheries, for cell shape and motility (lamellipodia, projections) -Microvilli -Cleavage furrow of cytokinesis -Sarcomeres

-Cell interior, for rigidity and vesicular transport -Mitotic spindles -Cilia, flagella

-Nuclear lamins, for nuclear structure -In cytoplasm for support -ECM (i.e. keratin) for mechanical support

Endocytosis and Lysosomes

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Endocytosis: General phenomenon of cellular uptake via mechanisms involving membranes (not channels or pumps) Pinocytosis (cell drinking) -Macro/micro Mediated by caveolin, causing cell invaginations  Non-specific, but not really random; tend to be positive/neutral molecules Phagocytosis (cell eating)  Can be specific (receptor-mediated) or non-specific  Contents go through endosomal system

Endosomal system  Consists of variable membranous structures (vesicles, tubules, MVBs), differing in shape and sizes  Involves progressive acidification of interior , initially by H+ pumps (pH goes down <6 ) -Traffic to lysosomes, which have low pH and hydrolytic enzymes >Enzymes are tagged with mannose-6-phosphate (M6P) in cis-Golgi - Receptors for M6P (M6PR) binds and sorts these enzymes at the transGolgi, bud as lysosomes to traffic to endosomes - Acidification causes cargo release, and M6PR receptors are recycled - Primary lysosomes are newly minted, have not combined - Secondary lysosomes have combined with endocytic cargo - Hydrolytic enzymes protected by glycosylation *If contents are undegradable, cell removes what it can and then condenses/sequesters material into a residual body -Progression can be generally categorized as early (near periphery) vs.late (near Golgi) but really is a functional continuum Ex: LDL receptor-mediated endocytosis  LDL recognized by receptor (LDLR) and these complexes will start to cluster  Clathrin, previously free in the cytoplasm, associates with these clustered receptors at their cytoplasmic ends

   

Coated pits form and pinch off to form vesicles. Clathrin disassociates and is recycled Acidification causes LDLR to release LDL, receptors are recycled back to plasma membrane Combines with primary lysosome, so that contents can be acted upon Vesicle membranes can be recycled

Heterophagy: uptake of external materials Autophagy: digestion of cellular components for turnover (proteins, ribosomes, mitochondria)

Clinical Correlates: Hypercholesterolemia: Defective receptor-mediated endocytosis of LDL (at the receptor level or trafficking level) causes extreme fatty deposits in various locations of body. -Premature atherosclerosis & heart attacks Infection -Many infectious agents use receptors as cellular entry points- have the ability to escape or neutralize lysosomal degradation >i.e. diphtheria toxin, almost every virus, parasites Inclusion cell disease  Defective enzyme responsible for tagging lysosomal enzymes  Lysosomal enzymes traffic through constitutive pathway out of the cell (secreted) Extra (?) Secretory lysosomes Conventional lysosome whose contents are also secreted Immunity -Cytotoxic T-cells release cytolytic proteins against cancer cells -MHC presentation of degradatory products

-Histamine from mast cells Albinism -Secretion of melanin from melanocytes

Peroxisomes and Mitochondria

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Lecture 6: Peroxisomes and Mitochondria Explain the role of peroxisomes in detoxification, catabolism, and biosynthesis. Peroxisomes (aka microbodies) producing and breaking down hydrogen peroxide Detoxification  Alcohol in liver and purines in kidney (abundant in organs) Catabolism  Oxidase breaks down long chain fatty acids, H2O2 side product Catalase degrades H2O2 H2O + O2 Biosynthesis  Gets membranes from the ER  Myelin precursor, bile acids, cholesterol Other notes: Replicate by fission Peroxins transport proteins from cytosol (receptor for PTS), important for biogenesis PTS (peroxisome targeting signals) at the C and N terminus on these proteins Diseases: Long chain fatty acid accumulation and low myelin Peroxisome biogenesis disorder (more severe) (eg: Zellweger syndrome: severe, peroxin mutations that lead to virtually no peroxisomes, fatal w/I 1st yr of life) Demyelination and accumulation of long-chained fatty acid Very flaccid and no muscle contraction Peroxisomal enzyme deficiencies (eg: X linked adrenoleukodystrophy, gene mutation of fatty acid transport into peroxisomes, death 1-10yrs after onset) Demyelination and fatty acid accumulation Lorenzo‟s oil 2. Explain the role of mitochondria in ATP production, catabolism, calcium homeostasis, and apoptosis. Mitochondria

ATP production: electron transport chain makes intermembrane acidic to drive ATP synthas (I) NADH dehydrogenase: H+intermebrane, e-ubiquinone (II) FADH2 dehydrogenase (succinate): FADH22H + FAD-+e-(ubiquinone) (III) Cytochrome C reductase: H+  intermembrane, e- (ubiquinone) cytochrome c (IV) Cytochrome c oxidase: H+ intermembrane, e- used to reduce O2 to water ATP synthase: H+ returns to matrix Catabolism: long chain fatty acid oxidation, pyruvateacetyl CoAoxidation Calcium homeostasis: sequesters excess cytosolic Ca (refer to muscles) Apoptosis: mito releases cytochrome c and activates a caspase cascade to induce apoptosis *NOT the same as necrosis (inflammation, uncontrolled release of cell contents) Other: Inner membrane is impermeable to small ions Tubular and can form networks Acetyl CoA Citric Acid cycle ETC (ATP synthase) Mt protein import (general): proteins contain mt targeting sequence Proteins bind the TOM and TIM (transfer protein of outer/inner mitochondrial membrane) and binding allows import into mito There are also secondary sequences which target inner membrane, intermembrane space, outer membrane Identify peroxisomes and mitochondria in electron micrographs. Compare and contrast the structure, function, and genetics of peroxisomes and mitochondria. Peroxisomes Structure 0.5-1μm Lipid bilayer; single membrane Granular matrix; oxidase, catalase Nonhumans have a nucleoid structure in the middle (big black spot) Mitochondria 0.5-10μm Inner membrane (cristae etransport) Outer membrane Intermembrane space (high H+) Matrix: mtDNA, ribosome,

tRNA, TCA enzymes Function Biosynthesis (bile acid, cholesterol, myelin) Detoxification of alcohol and purines β oxidation of fatty acids (very long chain) breakdown H2O2 Binary fission replication Membranes from the ER No endogenous DNA Disease biogenesis disorders are more severe Neurological myelination diseases Zellweger syndrome (no peroxisome; buildup of very long chain fatty acids) X linked adrenoleukodystrophy (no fatty acid transport into peroxisomes) ATP synthesis Ca homeostasis Regulation of apoptosis β oxidation of fatty acids

Genetics

Binary fission replication Inherited from egg (gamete) mtDNA (can have defects) autosomal mtprotein defects Mt enzyme deficiency Mitochondrial DNA defects Autosomal DNA defects for mt proteins Leber‟s hereditary optic neuropathy (loss of central vision, affects mostly males, optic nerve degeneration)

Identify maternal inheritance by pedigree analysis; explain how mitochondrial diseases can be maternally or autosomally transmitted; explain heteroplasmy. Maternal inheritance: Mitochondrial DNA defects (13 mt proteins, 22 tRNA, 2rRNA) Autosomal inheritance: nuclear DNA defects that code for mt proteins Heteroplasmy: mixture of mutated and normal mitochondria redistribute unevenly during development, so this is the occurrence of cells having different mitochondria and may occur within one individual Eg: Leber‟s hereditary optic neuropathy (loss of central vision, affects mostly males, optic nerve degeneration because not enough functional mito produce energy to maintain cells)

Microanatomy 7: Cell Differentiation 1. Describe the main characteristics of differentiated cells and understand that it is a result of differential gene expression, not gross changes in the genome itself. Differentiated cells are cells that have become morphologically/structurally, biochemically, and functionally specialized. Since nearly all cells of the body have genomic equivalence – i.e., they all possess the same full genome – cell differentiation typically results from changes in gene expression, not any changes in the DNA itself. Exceptions: Gametes (sperm and egg cells); immune system (B cells, T cells – splicing of immunoglobulin genes in order to generate antibody diversity) Once a cell is differentiated, the changes are often permanent and “heritable,” such that the cells produced through divisions of this differentiated cell have the same specialized characteristics that it does. Terminally differentiated cells (ex. neurons) lose their capacity for mitotic division entirely. In short, differentiation = restriction of cell fate 2. Describe the general properties of stem cells and compare them to differentiated cells. Properties of stem cells: - Capable of self-renewal or differentiation - May give rise to transit amplifying cells with limited division capacity - Often lack specialized organelles, and show high nucleus/cytoplasm ratio - Long-lived – express telomerase - Slow to divide, few in number

- May be restricted spatially to specific zones or niches - Respond to signals that will regulate their growth and proliferation Stem cells vs. differentiated cells seems pretty obvious just by extrapolating from the above properties, so a finer distinction to make might be stem cells vs. progenitor cells: Stem cells can divide an unlimited number of times, and can give rise to more stem cells (self-renewal) as well as differentiate into specialized cells of any of the three germ layers (pluripotency). Progenitor cells have a limited number of divisions and do not maintain their ability to self-renew, are considered to be at a further stage of differentiation than stem cells, and are more limited in the variety of cell types into which they can differentiate (multipotency or oligopotency). *Note: These two terms are sometimes used interchangeably, and the distinction between the two is still contested, since some stems cells (ex. adult stem cells) are also only multipotent, oligopotent, or unipotent. The best criterion to go by for distinguishing the two seems to be the selfrenewal aspect, which is exclusive to stem cells. 3. Distinguish determination and differentiation. Differentiation = the process by which different, specialized cells (morphologically, biochemically and functionally) arise from a homogeneous group of unspecialized cells. Determination = the commitment of a cell to differentiate into a certain cell type at some later time. Determination generally occurs as a step (or a series of steps) that is prior to, and separate from, differentiation. Cells can be determined a long time before they differentiate (at least by histological criteria). Two major ways that cells can become committed are by: 1) possession of special cytoplasmic determinants localized within the egg or early embryo, and 2) interaction with other cells or the factors secreted by other cells. Cytoplasmic determinants are components localized to a particular region of the egg or embryo that, when segregated to a cell or cell

lineage cause those cells to become determined, i.e., to adopt a particular cell fate.

First column: Normal pattern of differentiation; cells in region A differentiate into one type of cell, cells in region B into another type Second column: Sample of region B cells labeled and transplanted into region A early on, before they have become determined  transplanted cells differentiate into region A type cells Third column: Sample of region B cells labeled and transplanted into region A later on, when they are already determined  transplanted cells differentiate into region B type cells [Fourth column is not relevant to this objective, but is basically demonstrating that region B cells that are specified (specification = early, reversible/changeable step of determination, so not fully determined) will still differentiate into region B type cells in an isolated environment.]

4. Explain the potency of stem cells and understand it in the context of differentiation stages. Cell potency = amount of differentiation potential; the following are listed from least differentiated stage (have the most differentiation potential) to most differentiated stage (have the least differentiation potential) Definition Example

Can differentiate into Human zygote is any cell type (ectoderm, totipotent up through Totipotent mesoderm, and endoderm) as well as extra-embryonic tissue such as the placenta Can differentiate into any cell type (ectoderm, mesoderm, and endoderm), but cannot produce full embryo Can differentiate into a limited number of cell types Multipotent Embryonic stem cells (which originate from the inner cell mass of the human blastocyst) are pluripotent. Progenitor cells; also, hematopoietic stem cells (a type of adult stem cell) can differentiate into various blood cells (RBCs, white blood cells, platelets, etc.) but not other mesodermal cell types. Progenitor cells; some adult stem cells Skin cells the 8-cell stage.

Pluripotent

Oligopotent

Can differentiate into very few cell types Can differentiate into only one cell type; however, unipotent stem cells can still selfrenew

Unipotent

Terminally differentiated

Have fully differentiated Neurons and can no longer divide via mitosis

5. Explain inductive interactions and epigenetic controls in cell differentiation. Induction = process by which surrounding cell or environment guide the cell differentiate process; can be mediated through cell-cell, cell-matrix, or diffusible growth factor interactions; can be instructive (signal itself causes target cell to differentiate) or permissive (cell already committed, induction creates conducive environment for differentiation); timing of induction is crucial (see objective #6 on temporal specificity).

Epigenetic controls – contribute to maintenance of differentiation through heritable alterations in chromatin structure (ex. DNA methylation) and transcription factor expression. 6. Draw a diagram that demonstrates the principle of temporal specificity in induction.

7. List the sources of stem cells for use in research and medicine. Type Adult stem cells Pros Easily obtained; no controversy Pluripotent Cons Multipotent or unipotent, so have limited usefulness Political/ethical controversy; potential for aberrations leading to tumor formation during culturing stage Cord blood stem cells Mostly multipotent (peripheral blood stem cells, or PBSCs), but also some pluripotent stem cells; less (None mentioned in lecture)

Embryonic stem cells

controversial Induced pluripotent stem cells (iPSCs) Derived from nonpluripotent cell, ex. adult skin cells, so also no controversy; can be reprogrammed to regain pluripotency and even totipotency Chances of aberrations  tumor formation in origin cells, and during reprogramming and culturing stages; require more research

Intro to Tissues
Intro to Tissues Dr. Wood 9.1.2011 Generalities: All cells have: -Nuclei,EXCEPT RBCs or mitotic cells -Cytoplasm -Plasma membrane

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**They may be hard to see, due to abundance or plane or section Common cell shapes: Squamous Cuboidal Columnar Spherical Spindle Stellate When talking about tissues, don‟t forget to think about the ECM associated with the cells Epithelial tissue -Lines free surfaces, both externally and internally (i.e. skin, sweat glands, GI, kidney tubules) -Supported by connective tissue -Serves as barriers -Cells tend to be tightly packed (look for high density of nuclei) with little ECM -Avascular -Tend to be polarized Connective tissue -Provide structural support for many other types of tissue -Tend to have few cells (sparse density of nuclei) with lots of ECM

-Fibroblasts are the most common type of „connective cell‟ >Stellate/spindle shape, constitutively secreting ECM (collagen) -Classified as „loose‟ (more cells/water, less protein/fibers) or „dense‟ (more protein/fibers, less water/cells) -Blood is connective tissue -Also fat, bone, tendon, cartilage Contractile tissue -Specialized for rapid movement, responsive to electrical signals -Striated (cardiac, skeletal) vs. smooth muscle -Voluntary (skeletal) vs. involuntary (cardiac, smooth) -Many tubular structures (vasculature, GI tract) have contractile tissue *Smooth muscle can often look like connective tissue. Try looking at how many nuclei there are- more nuclei usually signals smooth muscle Nervous tissue -Neurons are large cells responsive for conveying electrical stimuli -Glia are smaller supporting cells (50:50 – 90:10) -Neuropil is cellular (not extracellular) material between cell bodies- cell processes of glia and neurons Origins of tissues from germ layers Ectodermal Origin Epidermis of skin Mesodermal Origin Dermis of skin Endodermal Origin GI (stomach, intestines, epithelial lining) but NOT including muscle layers Nervous system Cornea Lens Circulatory (heart, vascular epithelia) Kidney tubule epithelia Skeletal muscle, skeleton Connective tissue Lymphatic systems Excretory system Lungs and epithelial lining Liver Pancreas Bladder/urethral lining

(kidneys and bladder, excluding lining of bladder) Lining of body cavity (mesothelium?)

Epithelium

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1) Explain how epithelia establish barriers and control exchange between different environments Epithelium lines the free surfaces of the body, so material (ions, proteins, oxygen, sugars) needing to pass between tissues of the body and luminal spaces (lungs, gut, ducts) needs to pass through the epithelium. Epithelial cells are densely packed and connected to neighboring epithelial cells through: Tight junctions: forms a ring around the cell that maintains polarity of transport proteins between apical and basolateral faces. Not anchored by cytoskeletal elements Adhering Junctions: forms a ring around the cell, connected to actin Desmosomes: Spot welds connected to intermediate filaments This most commonly happens in a transcellular pathway, although there are examples of paracellular transport through the tight and adhering junctions. Because of cell polarity, different transporters on apical and basolateral surfaces can regulate directionality of transport and create downhill gradients across the cell. The apical side of epithelium in the trachea are coated in cilia that facilitate transfer of mucus out of the airway Keratinized stratified squamous epithelium provides a particularly strong barrier with a thick layer of dead, denucleated squamous cells that are continuously regenerated as they are sloughed off. 2) Classify Epithelium based on cell layers and cell morphology Simple Squamous Epithelium: fried egg morphology, can have elongated cytoplasm. (endothelium, alveoli, mesothelium, portions of kidney tubule) Simple Cuboidal epithelium: cuboidal, regular (or just not columnar or squamous) morphologyshape. (kidney tubules and ducts) Simple Columnar Epithelium: Tall cells, usually with apical surface specialization (gut, intestine, respiratory tract, oviduct

Stratified Squamous Epithelium: Only top “living” layer has to be squamous. Usually serves to protect underlying tissue. Can be keratinized (skin cells) or non-keratinized (mouth, vagina, esophagus) Transitional Epithelium: Stratified epithelium that lines urinary system from center of kidney (Renal calyx) to bladder. Allows stretching. - rounded (bumpy) surface is identifying characteristic Pseudostratified epithelium (Respiratory Epithelium): All cells contact the basement membrane, but not all are exposed to the lumen 3) Explain replacement of epithelium: Determined cells can undergo mitosis to form new cells (liver, endothelium) or stem cells can undergo mitosis and determination (intestines, skin). In stratified epitheliums, the new cells are generated near the basement membrane and migrate to the top as cells are sloughed off. 4) Explain different types of glandular secretion: Merocrine: secretory granule fuses with the PM and dumps it contents into the ECF Apocrine: secretory granule leaves cell where its contents are still enclosed in plasma membrane (i.e. mammary glands secreting milk) Holocrine: Whole cell lyses and dumps contents (ex/ sebaceous gland) Paracrine: Cell releases contents that then signals nearby cells. Commonly, this occurs with signaling molecules that degrade quickly and can thus only act on nearby cells. Autocrine: Cell releases contents that then binds to same cell (B-cells)

Cell-cell/Cell-matrix interactions

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Multicellularity allows cells to specialize and carry out more tasks simultaneously To become multicellular: - Contact and communicate with other cells - Regulate passage of environmental substances into and out of organism - Recognize self/non-self Occluding/tight junctions: do not involve cytoskeletal elements Formed by claudin and occludin (transmembrane proteins) associations Form tight seal that that not allow most molecules to pass through Found usually on apicolateral surface of cells (epithelial usually) Anchoring junctions: do involve cytoskeletal elements Actin Microfilament associated Cell-cell junction (mediated by cadherins) Cell-matrix junction (mediated by integrins) Focal adhesions Hemidesmosomes Adherens junctions Intermediate Filament associated Desmosomes

Homophilic vs. heterophilic binding Cadherin family: Calcium-dependent adhesion molecules Low Ca++: cadherins are largely disorganized ~1mM Ca++: Dimers form extracellularly, nucleates other dimers – „molecular zipper‟ Adherens Junctions: E-, N-, VE-cadherins („classical‟) Desmosomes: Desmocollin, desmoglein

Linked to actin cytoskeleton plakoglobin/desmoplakin via α/β catenin

Linked to IF via

*Cadherins mediate cell-cell sorting where cells expressing the same cadherins (hemophilic binding) associate with each other and organize into structures Other adhesion molecules: Ig superfamily: Ca-independent family of proteins that are weaker than cadherins and can be both homophilic and heterophilic Selectins Ca-dependent group of adhesion molecules that bind oligosaccharide lectins (heterophilic) Low affinity binding. Responsible for leukocyte „rolling‟ ECM Hydrated, polysaccharide gel substance with fibrous proteins embedded, important for physical stability and diffusion of important substances. Usually formed via fibroblasts. The basal lamina is a specialized form of ECM laid down by epithelial cells. ECM components Glycosaminoglycans (GAGs): Unbranched, repeating disaccharide units. Hydrophilic, highly negatively charged- responsible for resisting compressive forces Ex: hyaluronan Proteoglycans: Protein + GAG = proteoglycan Formed by a tetrasaccharide link Structral and cell-cell signaling roles

Fibrous proteins: Collagen – long inflexible fibers providing strength Elastin - mesh-like fibers that provide flexibility Fibronectin – recognized by attachment proteins Laminin - recognized by attachment proteins Integrins Transmembrane receptors (heterodimeric) with affinity for RGD motif in fibronectin. Linked to actin/IF cytoskeleton via talin. Can communicate bi-directionally. ECM →cell: Leukocyte recruitment Cell → ECM: ECM remodeling for „pathfinders‟ Communicating junctions: mediate chemical or electrical signals from one cell to another Gap junctions Mediated by membrane channels called connexons, which are formed by 6 connexin proteins Permeability affected by pH and Ca++ (open with high pH, low Ca++) Chemical synapses Fast type of paracrine signaling

Blood
Element Function Identifying characteristic

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Lineage
proerythroblasts -> erythroblasts ->

no nucleus or other transport O2, transport a little Erythrocyte CO2 trigger thrombi to limit blood loss at injury site; aggregate w/ platelets, alter blood flow, initiate Thrombocyte (platelets) coagulation cascade phagocytosis & destruction of Neutrophil bacteria destroy larger parasites / modulated allergic Eosinophil response release histamine & heparin (anticoagulation Basophil ) mature into macrophages -Monocyte > phagocytosis S-shaped nucleus, blue/black granules irregular, kidneyshaped nucleus; no granules 4-10% <1% multi-lobed (2); pink/orange granules 1-3% multilobed; clear or light pink granules 60-70% discoid, small fragments (small specks interpsersed w RBCs) many organelles, biconcave shape, appear electron dense most abundant

reticulocytes (no nucleus) -> RBCs (left bone marrow, no more RNA) megakaryoblast >megakaryocyte (in bone marrow) --> 1000 platelet clumps (made of cytoplasm)

myeloid cell myeloid cell

myeloid cell

lymphoid cell: enter lymphoid organs --> macrophages (Myeloid lineage)

B cells --> plasma cells; T cells --> Lymphocyte immune rxns produce platelets; located in bone Megakaryocyte marrow produce RBCs; located in bone Reticulocytes marrow found in connective tissue; secrete targeted Plasma cells antibodies found in connective tissue; phagocytic; act as antigenMacrophages presenters many cytoplasmic processes, numerous lysosomes lots of rough ER that is dilated; clumps of heterochromatin; no storage vessicles largest cell; has clusters of proplatelets look like RBCs but still have RNA (blue stain in cytoplasm) large, round, dark nucleus 20-30%

myeloid cell: B cells -> plasma cells

Remember: Never Let Monkeys Eat Banas

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