SPECIALIZATIONS OF THE CELL SURFACE MICROVILLUS, CILIA, FLAGELLA Prof Dr Müjgan Cengiz İÜ.Cerrahpaşa Tıp Fakültesi Tıbbi Biyoloji AD The surface of the most cells have extensions They are used in cell movement, phagocytosis, absorbtion. Most of these extensions are based on actin filaments. I- SPECIALIZATIONS OF FREE SURFACE (APICAL SURFACE ) 1-Microvilli 2-Cilia, flagella 3-Stereocilia 1-Microvilli Finger-like extensions derived from the cell surface Function: increase the surface area for absorption, cell movement and phagocytosis Localisations( cells specialized for absorbtion ) 1-İntestinal epithelium (Striated border ) 2-Proximal tubule of the kidney (Brush border) 3- Gall bladder epithelium Microvilli Finger-like prolongations 1 in length 0,1 diameter (can be seen with L.M.) It can seen with E.M. Contains a bundle of straight parallel filaments (20-30 Actin filaments ) Actin filaments extend 0,5 down into the apical cytoplasm Enclosed in an extension of the plasma membrane Glycocalix is ticker around the microvilli Microvilli (Actin based cell surface protrusions) A bundle of actin filaments are cross linked into closely packed parallel arrays “Actin binding proteins” cross link actin filaments ; Fimbrin, Villin Striated borders (microvilli ) contain enzymes i.e sucrase maltase lactase lipase aminopeptidase They are involved in the terminal digestion of proteins, carbohydrates and lipids 2-Cilia (kinocilia ) and Flagella Cilias have eyelash or hair like structures -Motile -Larger than microvilli (5-10 long , 0.2 in diameter) -Have a complex internal structure (E.M ) 250 or more cilia (in each cell) arranged in parallel rows Localisations: 1-Epithelial cells of the upper respiratory tract 2- Epithelial cells of the uterine tubes (oviducts ) 3- Epithelial cells of the Ductus efferentes Ciliary movement is constant in direction In living cells Cilia beat in a rhythmical wave –like manner (phase-contrast microscope) Functions: 1-to move mucus and particles over epithelial surfaces ( in respiratory epithelium ) 2-to transport the ovum toward the uterus (in uterine tubes) 3- to drive the spermatozoa toward the epididymis (in ductus efferentes ) Cilia With L.M Hair –like protrusions No internal structure is detectable At the base of each cilium a dense granule (Basal body) is seen With E.M Have a complex internal structure A characteristic arrangement of microtubules called “ Axoneme’’ =9+2 microtubule complex Fawcett 1954 Cilia Each cilium is covered by an extention of plasmalemma 1-Tapering tip 2-Cylindrical shaft 3-Basal body (located in the apical cytoplasm) Dynein arms are arranged along the length of the microtubule . They are formed by a protein called “ dynein’’ and contain ATPase (ATP splitting enzyme)activity. “Nexin links’’ attach each microtubule A to the microtubule B of the adjacent doublet. Nexin links are composed of an elastic material called “nexin’’ Responsible for recovery stroke Basal Bodies Cylindrical structures about 0.2 in diameter and 0.4 in length nine groups of three microtubules fused into triplets form the wall of the basal body Basal body resembles a centriol basal body contains some accessory structures such as rootlet and basal foot Accessory structures anchor the cilium in apical cytoplasm MECHANISM OF CILIARY MOVEMENT Cilia beat in a rhythmical wave-like manner’’ old concept: Ciliary movement is based on the contraction of microtubules (Microtubules are capable of shortening ) Satır, examined cross sections near the tips of cilia in different phases of beating Doublets of the bend cilium terminate at different levels Sliding Microtubule Mechanism (Satır) If a cilium is straight doublets terminate at the same level If a cilium is bent toward doublets 5 and 6 5 and 6 project farthest Doublet 1 terminates first If movements were produced by microtubule shortening, microtubules on the concave side (5 and 6) Should be shorter . Result: Bending occured without shortening Sliding Microtubule Mechanism (Satır ) To day accepted mechanism: A cilium bends a long the axis by a type of ” Sliding Microtubule Mechanism’’ between microtubules It is similar to that seen between myofilaments in striated muscle fibers Kartagener’s syndrome; 1-Chronic sinusitis 2-Bronchiectasis (chronic dilation of bronchi ) 3-Situs inversus totalis 4-Male infertility Normal number of spermatozoa but no motility Afzelius 1978 E.M Afzelius’s Experiment 1978 Electron Microscopic observation of immotile sperm flagellum of patients with Kartagener’s syndrome: EM observation of bronchial biopsies Showed the absence of dynein arms showed no dynein arms in ciliary axonemes Result: *Dynein is essential for motility of cilia and flagella Kartagener is a genetic disease Dynein is essential for motility of cilia and flagella Afzelius examination 1978 *Clinical and *Electron Microscopic observation on a congenital form of human infertility showed that dynein is essential for motility of cilia and flagella DYNEİN • Extremely large protein • Dynein arms form temporary cross bridges between microıtubule A of one doublet and microtubule B of the adjacent doublet •During sliding they undergo a cyclic break and reattachment •Formation of cross bridges is ATP dependent Dynein –Walking Model • Dynein appears to “ walk’’ along the adjacent doublet Experiment: • Proteolytic enzymes digest nexin links and radial links • The addition of ATP produces a sliding movement of the bridge along the B tubule of the adjacent doublet • Doublets move relative to each other powered by the motor activity of axonemal dynein • Radial linkers convert the sliding of microtubules into bending of cilia Ciliary movement Requires 1-ATP 2-Ca ions , Mg ions STRUCTURE OF MICROTUBULE Microtubule (13 protofilaments ) Proto filaments are composed of Tubulin subunits (dimer) Dimer→ Tubulin α ve Tubulin β Flagella (Same internal structure with cilia) (Axoneme 9+2 ) Long whiplike protrusion whose ondulations drive a cell through a fluid medium. Eukaryotic flagella are longer than cilia. Bacterial flagella smaller and different mechanism of action. FLAGELLA 1- Longer than cilia (100-200 μ ) 2-Different type of movement (undulating wave type of movement) 3- Less in number (one or two in a single cell) 4- Mammalian spermium contains 9 additional dense fibers arround the axoneme (9+9+2) (protective function) Stereocilia 1-Long and irregular microvilli 8μ (EM) 2-Have no internal structure 3-Microfilaments are poorly developed 4-No motility 5-No basal body Function: Increase the cell surface for absorbtion Localisation: Ductus epididymis, ductus deferns