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The cell Histology Is the microanatomy of the cell and tissues, using different histological methods of staining guided by LM and TEM Cytology = is the study of the cell structure and function Every cell consists of protoplasm surrounded by cell membrane The mass of the protoplasm is divided into two components: o Cytoplasm which lies between the cell membrane and nuclear membrane, it constitutes the main bulk of protoplasm o The nucleoplasm which fills the nucleus. These plasms are colloid or (Semi fluid) and fluctuate between gel and sol conditions. Cytoplasm is not stationary. It is a state of flux i.e. organelles can work quite and in autonomous manner as well as actual movement of water and ions. Cells divide into two major compartments o Cytoplasm o Nucleus Suspended within the cytoplasm - organelles - Inclusions Organelles They are specialized parts of living substance and probably Always present They are divided into membranous and non-membranous Organelles The cytoplasmic ground substance was called cytosol now called cytoplasmic matrix. Copyright@ 2003 pearson Education. Inc., publishing as Benjamin Cumming Membranous organelles That is surrounded by membrane: Plasma membrane. Mitochondria Endoplasmic reticulum sER Golgi apparatus Lysosomes Endosomes Peroxisomes rER Non membranous organelles include: Microtubules These form the cytoskeleton of the Cell = (Cytoplasmic support) NB Filaments Centrioles Ribosomes Some references refer to ribosomes as membranous organelles Inclusions: They are materials often of a temporary nature and present under special conditions. (1) Membranous organelles Plasma membrane: Cell membrane = plasma lemma It participates in many physiological and biochemical activities essential to cell survival and function. LM: It is not visible EM: It displays a characteristic trilaminar appearance that has been described as a unit membrane. Trilaminar i.e. two electron dense lines (inner & outer) and a clear or lucent single line in between (intermediate) Diameter: 7 – 8 nm Molecular composition: It mainly consists of lipid and protein. Lipid: Consists of a bimolecular leaflet of phospholipids which is the back bone of the cell membrane The fatty acid chains in the phospholipids facing each other with their hydrophilic heads directed to the exterior and their hydrophobic tails directed to interior surface. The Cell Membrane The Cell Membrane (Schematic view of the neuronal cell membrane) (The Eaton T. Fores Research Center) Protein molecules Fluid mosaic model like iceberg float in a sea of phospholipids, it constitute about 60 – 70% of membrane mass. It is described within lipid by two ways: (1) Integral protein = contain hydrophobic + hydrophilic regions are inscribed into the lipid bilayer and extend completely or partially. They attach firmly to the phospholipids only drastic measure as using of detergent can release them. (2) Peripheral protein = are external to lipid bilayer. They are attached to the head by week bonds which easily break by changing PH Function: Participate in enzymatic activities. Maintain ionic conc. across to the opposite sides of the membrane. Receptor protein recognizes the binding of substances (important in hormones, antibody reaction). For reading only According to their function, there are six categories of Proteins (e.g. pumps channels, Receptor, transducer carrier, structural) Read about: cell signaling. Cholesterol: Intracellular membrane less cholesterol It is associated with fatty acid tails and changes their molecular nature i.e. fatty + cholesterol acid determines whether the lipid tail is crystalline or loose (fluid). more cholesterol present, the more fluid (loose) the membrane will be. Carbohydrate: (glycocalyx = cell coat) It is linked either to protein or lipid and present in the extra cellular surface. Link to protein glycoprotein Link to lipid glycolipid These surface molecules constitute a layer at the surface of the cell called cell coat or glycocalyx. Function Act as cell recognition and cell adhesion. Protect the cell against chemical injury. Act as specific receptor sites for incoming stimuli such as hormones. Reading sector - (Read about theory of cancer and graft rejection) Homework: Function of cell membrane (read it !) 1- Selective barrier 2- Exchange of materials between in + outside the cells. 3- Conduct impulse such as nerve cells. 4- Cell recognition + adhesion. 5- Receptor sites are selective to stimuli. Endocytosis, phagocytosis, pinocytosis, receptor mediated Endocytosis, exocytose. (2) Mitochondria It is a membrane limited organelle. They act as the chief source of energy in the cell. Structure They differ in their numbers and shapes. Under phase contrast microscope: they appear like threads that are about 0.2M and several micrometers in length. They can divide by binary division i.e. self replicated; because they contain DNA loops, particles resemble ribosomes and RNA molecules; therefore, they can also synthesis their own protein. EM: Mitochondria consist of a smooth outer membrane and highly folded inner membrane convolutes into cristae that project into the matrix. The space between the two membranes is called outer compartment. Matrix is the inner compartment. Mitochondria The inner surface of the cristae that faces the inner compartment possesses elementary particles. Each consists of a head projects into the matrix and a stalk that attaches to the inner membrane. Matrix contains mitochondrial granules, DNA loops and RNA particles. Function It is the energy source of the cell utilizing pyruvate and generates ATP by the process of oxidative phosphorylation. They are found in large amount in muscle Matrix granules contain enzymes of kreb's cycle Respiratory chain enzymes are in the inner mitochondrial membrane Elementary particles contain the enzyme of the electron transport system and AT pase enzyme. They control the conc. of certain ions especially ca++ and release it into the cytoplasm when needed. (3) Endoplasmic reticulum They are branching network of tubules There are two types: rough (rER) and smooth (sER). Smooth endoplasmic reticulum (sER) Structure: They consist of a complex system of anastomosing short tubules that are associated with ribosome's. LM: cells, which are rich in sER, have a distinct acidophilia Function: It has a despair function in different cells. FOR EXAMPLE o SER is very well developed in cells that synthesize and secrete steroid hormone, a precursor of steriodogenesis (cholesterol) is stored in SER. Site: sex organs (testicular interstitial cells) and adrenal cortical cell. o Striated muscle (skeletal + cardiac muscle) possesses highly developed system of SER called sarcoplasmic reticulum. Ribosomes and rough endoplasmic reticulum It helps in uptake and releasing of ca++ ions which are essential in contractile process of the muscle. o sER of the liver hepatocyte contain enzymes that are responsible for detoxification of lipid soluble chemicals e.g hydroxylated enzymes. sER may involve in lipid absorption. Ribosomes and rough endoplasmic reticulum They are abundant in the cells that produce protein for secretion. LM: the cells rich in rER and ribosomes stain intensely with basic dye (basophilic cells) TEM: rER appears as a serious of branching and interconnecting Membrane limited flattened sacs or saccules that called cisternae. - Cisternae are closely packed in parallel rays. - The outer membranes of these cisternae facing cytoplasm are studded with granules called ribosomes. - Saccule of rER usually continues with the outer nuclear membrane. (4) Ribosomes They are granules that consist of two subunits which are large and small subunits with different sedimentation coefficient. They attach to the rER by their large subunit and contain RNA and protein. They present in either free or attached form. The free ribosomes circulate in the cytoplasm. There are two types of the attached ribosomes: 1- Ribosomes attached to his side of rER that face the cytoplasm. 2- Polysomes or polyribosomes: Several ribosomes attached to a thread of RNA. Function RER responsible for protein for destination (secreted outside the cells) and cell membrane integrity. Free ribosomes are responsible for protein synthesis that is used inside the cell. TEM: Transmission electron microscope Ribosomes translate the genetic message from mRNA that Determine the sequence of amino acids for particular protein. These proteins will be elaborated immediately to the 14 men of rER. RER give off buds of ribosome free vesicle (transfer vesicle), so the protein has been segrated by those vesicles more to ward the immature surface of golgi saccule. Golgi apparatus It is discovered by Golgi in 1898. It is usually located between rER and plasma lemma. LM: - The position f golgi complex appears (using H&E) unstained area near the nucleus while the whole cytoplasm is stained (Basophilic). This is called negative golgi image. - By silver or lead or osmium staining, golgi complex appears as fibril or granular network. These network appear as brownish or black network supranuclear in position. Site: Highly present in cell secret protein e.g. acinar cells of Pancreas and epidemics. EM: Golgi is membrane bounded organelles formed of a stack of: -Parallel flattened cisternae with smooth surface membrane. They associated with vesicles. Each cisternae has two sides or faces. They are composed of: a- Cisternae or saccules: A, 1- Cis – Golgi or forming face (convex side) facing rER A, 2- Trans – Golgi or maturing face ( concave side) that face the plasmalemma. b- Transport vesile: carry newly formed protein from rER. c- Condensing vacuoles: that pinch out from the trans-face containing protein ready for secretion. Function Golgi apparatus concern with secretory activity of the cell and processing the secretory product in one or more of the following ways: 1- Glycosylation and sulfation of glycoproteins and glycolipid. 2- Proteolytic processing of presecretory protein. 3- Concentration, packaging and sorting of the secretory product into membrane secretory vesicles. 4- Membrane biogenesis though the process of fading of secretory vesicles. Protein synthesis The formed protein polypeptide chain in rER will be segregated and transfer toward the Golgi complex through transfer vesicle then fuse with membrane, subsequently release to the lumen and pass through cisternae toward the mature face (trans – Golgi). Within Golgi carbohydrate is added (glycosylation) glycoprotein or sulpha group is added protoglycan is formed. - The enzymes responsible for these processes are found in the membrane of Golgi saccule. - Then the protein will be secreted by condensing vacuole which a bud from the dilated rim by transface vacuole. - In the vacuole will be concentrated and vacuoles become smaller and called secretory vesicles. - It will pass to toward the plasmalema, fuse within it and release its content to outside. Two types of protein could be release for golgi: 1- Lysosomal enzyme 2- Protein for secretion Perixosomes: Microbodies They are small membrane limited spherical (0.5Mn) bodies that contain oxidative enzymes involve in hydrogen peroxidase metabolism. They contain catalase and other peroxidase that break down hydrogen peroxide. (H2O2), (toxic substance). They are present almost in most of the cells but they are numerous in liver and kidney cells. The number of peroxisomes present in the cells increases in response to diet, drugs and hormonal stimulation. Endosomes = They are product of phagocytosis. Lysosomes LM: They are of visible membrane bounded vacuoles 0.2 – 0.5Mn EM: They have heterogeneous morphology They contain hydrolytic enzymes responsible for digestion of intra or extra cellular substance They present in almost all cells except erythrocyte. Types a- Primary lysosomes: these are lysosomes that recently formed from Golgi apparatus and doesn't contain digested material b- Secondary lysosome: when primary lysosome fuses with membrane of structure that contains the material to be digested and release their enzyme. They either called phagosomes or digestive vacuoles or autophagic vacuoles depending on (the intra- or extra-cellular) material to be digested. Example of phagosome = any bacteria e.g of autophagic vacuole = old mitochondria. If secondary lysosome (phagosome) digests its content residual body will be formed exocytosis to expel the digested material. Pinocytotic vesicle when fuse with lysosome, it is called multivesicular body. In nerve and cardiac cells there are accumulation of residunal body (can’t be expelled outside the cell) to form lipofucsin granules or age pigment a golden brown pigment. It is a normal feature of aging. Diagram of the basic architecture of a cell. Note the location of the Rough Endoplasmic Reticulum and the Golgi apparatus within the cell. Taken from: Junqueira and Carneiro, Basic Histology, Text and Atlas, page 42, Figure 2-27. Molecular Biology of the Cell (Hardcover) Secondary lysosome (red arrow) and peroxisomes with lamellar nucleoids (blue arrows) in the pyramidal cell. N - nucleus. Scale = 400 nm. (Rat, hippocampus.) Atlas of Ultrastructural Neurocytology (Josef Spacek) Reading section Lysosomal storage disease = where accumulation of residual bodies in the cells result in interference with normal function of the cells, .g. Tay sachs disease (nervous tissue). Mucopolysaccarydosis. Cytoskeleton They are interna cytoplasmic support system that maintains control of the cell shape. It is mainly include: 1- Microtubules. 2- Thick filaments. 3- Intermediate filaments. 4- Thin filament microfilaments. Microtubules They can’t be seen by LM unless if they are present in thick bundles. They are non-branching – hollow cylinders that measure 20 – 25nm in Diameter they are composed of tubulin diamers. They are labile structure that can change by polymerization and depolymerization. Microtubules found in most of the cells but particularly: 1- Axonema of cilia, flagella. 2- Basal bodies of cilia 3- Mitotic spindle. 4- Centrioles from which spindle fibers radiate. 5- Growing axons 6- Cytoplasm in general. Microtubules involved in numerous essential cellular activity that relate to cytoskeletal function including: 1. Cell elongation and movement (migration). 2. Intracellular transport of secretary granules. 3. Movement of chromosomes during mitosis and meiosis. 4. Maintenance of cell shape. 5. Beating of cilia and flagella. Cytoskeleton Microtubules Microtubules with dark centers (arrow) in a small dendrite. Scale = 100 nm. (Mouse, cerebellar cortex.) (Atlas of Ultrastructural Neurocytology - Josef Spacek) Filaments They are as following: 1- The microfilament Actin = thin filaments – in all cell types on plasma membrane Myosin = thick filaments in muscle cell Tropomyosin these all responsible for muscle contraction, spectrin (in RBC). Myofilaments microfilaments have role in wound healing. 2- Intermediate filaments 1. tono filaments in epithelial cells. 2. Neurofilaments in nerve cells. 3. Glial filaments in glial cells. 4. Desmin filaments in muscle cells. 5. Vementin filaments in mesenchymal cells Cell membrane specialization Apical microvilli, cilia cell coat (gylocalyx) Lateral junctional complex gap junction (nexus) Basal basement membrane Hemidesmosomes Centrioles: Usually occur in pairs each called centrosomes In non-dividing cell are present near the nucleus. They consist of nine triplets microtubules. The centriole and unknown materials surrounding it could be microtubules forming microtubules organizing center It has important role in cell division and microtubules production. Microvilli: Are projections of plasmalemma. Present in certain epithelial calls for absorption They contain thin filament (actin) which are the structure core of microvilli at the apex they attach to a dense area dense lip at the base embedded the actin filament in to terminal web which also contain myosin. These microfilament responsible for the movement of microvilli e.g. brush border of small intestine . Cilia Are invagination or finger like projection of cell membrane found only on free surface of cell that lining lumens or cavities. They facilitate movements of fluid e.g. R.S Each cilium has basal body, shaft and rootless Basal body is identical to structure of centriole and it is the center of production of cilia. It is composed of two central (pair) microtubules surrounded by nine microtubules doublets. A radial spoke radiates from each doublet to the central pair of microtubules. Each doublet has two short arms attach to adjacent doublet they are formed from dynein that possess ATpase activity the hydrolysis of ATP give the energy to generate the cilia beating movement of flagella similar the cilia and present in the tails of the sperm. The nucleus It is the storage of genetic information in DNA In non-dividing cell called interphase. It is composed of nuclear envelope nucleoplasm chromatin and nucleolus. It has affinity to basic dyes to it its rich content of DNA. Usually spherical or ovoid. Nuclear membrane It separates the nucleus from the cytoplasm It has 3 structure: a- Inner nuclear – usually fibrous (adjacent chromatin) outer nuclear usually rough b- Nuclear pore complexes. c- Nuclear lamina. Inner + outer membrane separated by a space called perinuclear cisternae which is continuous with lumen of rER. Chromatin Consisting of strands of DNA and its associated protein. It is responsible for characteristic basophilia of the nucleus. They are not homogenous structure. - Highly condensed chromatin = Heterochromatin which are predominate in less active cell deeply stained. - Extended (highly stained) or dispersed euchromatin in high active cell. Heterochromatin present in the location of Marginal – re – inner nuclear membrane Karyosome irregular bodies Nucleolar associated chromatin in association with nucleus dead cell = nucleus usually shrunked integral and densely shrind all chromatin is heterochromatin pyknotic cell. Nucleolus = is non membranous intra nuclear structure. In LM: most cells have one or more nucleoli they are ovoid in shape and stain with basic dye. It is responsible for ribosomal RNA synthesis. In EM = it consist of 2 pairs 1- Pars granulose = nucleolonema = transcribed ribosomes. 2- Pars fibrosa (fibrillar). Inclusions Pigment Melanin in melanocytes in the iris of the eyes Golden brown pigment = lipofuscin. Haemosidrin = hemoglobin of aged RBC has been with age escaped to circulation + phagocytosed. Lipid Usually lipid droplets dissolve in preparation of tissue for LM and appears as round holes under LM. In EM = Appear black + spherical Glycogen = it can be demonstrated by periodic acid Schiff reaction appear as magentacolour. In EM = 2 particles identified: 1- α free part 2- β rosette Cell junction That bind cell to one another to form tissues. 1- Tight junction = zona occludens = built desmosome A minute or thread like constituent of plasmalemma crosses around the contiguous layer. EM: 2 membranes appear sealed Gap junction = nexus: allow ions and very small molecular Weight substances to pass 2nm space between the adjacent plasmalemma in between them. the nucleus takes up nearly 10% of the volume of the cell. Microsoft Encarta Picture http://www.geocities.com/auroranex/body_index.html