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Intracellular organelles and molecular mechanisms of organi z ation of  a  eucar y otic cell
T ypical organelles in eucaryotic cells
Membrane-bounded compartments  ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
  How to study organelles: optical and electron microscopy   Saccharomyces cerevisiae. Cell size : 3-5  µ m   Phase contrast microscope ELM – freeze -fracturing   ELM  -   u ltrathin section
How to study of organelles: differential centrifugation
How to study of organelles: gradient centrifugation
Nucleus Jaderný obal Jadérko Chromatin Nuclear  envelope Nucleus
Mitochondria C ompartment s: Outer membrane Matrix Intermembranous space Fluorescing mitochondria Mitochondria – ultrathin section
Chloroplasts Light microscope  E lectron microscope   Flattened membrane vesicles  acumulating to  form  grana grana
Endoplasmic reticulum flattened membrane cisternae  rough ER – cisternal surface covered by ribosomes  smooth ER cisternal surface without ribosomes Function:  rER-  s y nthesis of proteins for  secretory pathway  sER – synthesis of lipids and steroids
Golgi ap p ar a t us stacks of flattened cisternae with  perifer al vesicles Synthesis and packaging  of molecules destined to be secreted from the cell
Vesicular transport :  secretory vesicles (dark blue) and endosomes (light blue)
Peroxisomes small membrane-bounded vesicles  that provide containers for reactions where a dangerously reactive hydrogen peroxide is generated  and degraded.
Arrangement of organelles in a liver cell (multipolar) and in a pancreatic cell (polar orientation) Multipolar organization Polar organization
Cytoskeleton as a scaffoulding for cell organelles  Actin  Microtubules  Intermediate   filaments
Topology of organelles in a typical eucaryotic cell:  A cytoskeleton – a system of protein filaments (microtubules,actin filaments and intermediate filaments)   crisscrossing the cytoplasm and forming with other proteins a scaffolding for membrane organelles
The placement of organelles by microtubules green- MT blue – ER, yellow - GA MT ER MT GA
Amino acid sequences as signals for  r ecognition or  attachment
Attachment of  melanosomes to MT, actin filaments and PM Attachment of vaculoles to actin cables  via signal proteins and adaptors
Microtubule-based transport and intra-cellular trafficking pathways .  Many intracellular trafficking pathways involve active and directed transport along the microtubule cytoskeleton. Microtubule-dependent trafficking in mammalian cells includes:  (a,b,c)  ER-to-Golgi transport,  (d)  TGN-to-ER transport and  (e)  lysosomal,  (f,g)  endosomal and  (h)  mitochondrial motility
Plectin molecules serve as linkers between intermediate filaments (orange), microtubules (red) and actin    filaments (yellow)
Key terms from the lecture   1. Size of cells and organelles 2. Overview of eucaryotic cell organelles  3.Techniques of organelle separation and description (microscopy, gradient centrifugation,  differential centrifugation  4. Organelle description: nucleus, mitochondria, chloroplasts,  ER, GA,  secretory vesicles, lysosomes, peroxisomes   5. Cytoskeletal components and localization of organelles 6. How to prove co-localization of ER or GA with the microtubules

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L03 organeles

  • 1. Intracellular organelles and molecular mechanisms of organi z ation of a eucar y otic cell
  • 2. T ypical organelles in eucaryotic cells
  • 3.
  • 4. How to study organelles: optical and electron microscopy Saccharomyces cerevisiae. Cell size : 3-5 µ m Phase contrast microscope ELM – freeze -fracturing ELM - u ltrathin section
  • 5. How to study of organelles: differential centrifugation
  • 6. How to study of organelles: gradient centrifugation
  • 7. Nucleus Jaderný obal Jadérko Chromatin Nuclear envelope Nucleus
  • 8. Mitochondria C ompartment s: Outer membrane Matrix Intermembranous space Fluorescing mitochondria Mitochondria – ultrathin section
  • 9. Chloroplasts Light microscope E lectron microscope Flattened membrane vesicles acumulating to form grana grana
  • 10. Endoplasmic reticulum flattened membrane cisternae rough ER – cisternal surface covered by ribosomes smooth ER cisternal surface without ribosomes Function: rER- s y nthesis of proteins for secretory pathway sER – synthesis of lipids and steroids
  • 11. Golgi ap p ar a t us stacks of flattened cisternae with perifer al vesicles Synthesis and packaging of molecules destined to be secreted from the cell
  • 12. Vesicular transport : secretory vesicles (dark blue) and endosomes (light blue)
  • 13. Peroxisomes small membrane-bounded vesicles that provide containers for reactions where a dangerously reactive hydrogen peroxide is generated and degraded.
  • 14. Arrangement of organelles in a liver cell (multipolar) and in a pancreatic cell (polar orientation) Multipolar organization Polar organization
  • 15. Cytoskeleton as a scaffoulding for cell organelles Actin Microtubules Intermediate filaments
  • 16. Topology of organelles in a typical eucaryotic cell: A cytoskeleton – a system of protein filaments (microtubules,actin filaments and intermediate filaments) crisscrossing the cytoplasm and forming with other proteins a scaffolding for membrane organelles
  • 17. The placement of organelles by microtubules green- MT blue – ER, yellow - GA MT ER MT GA
  • 18. Amino acid sequences as signals for r ecognition or attachment
  • 19. Attachment of melanosomes to MT, actin filaments and PM Attachment of vaculoles to actin cables via signal proteins and adaptors
  • 20. Microtubule-based transport and intra-cellular trafficking pathways . Many intracellular trafficking pathways involve active and directed transport along the microtubule cytoskeleton. Microtubule-dependent trafficking in mammalian cells includes: (a,b,c) ER-to-Golgi transport, (d) TGN-to-ER transport and (e) lysosomal, (f,g) endosomal and (h) mitochondrial motility
  • 21. Plectin molecules serve as linkers between intermediate filaments (orange), microtubules (red) and actin filaments (yellow)
  • 22. Key terms from the lecture 1. Size of cells and organelles 2. Overview of eucaryotic cell organelles 3.Techniques of organelle separation and description (microscopy, gradient centrifugation, differential centrifugation 4. Organelle description: nucleus, mitochondria, chloroplasts, ER, GA, secretory vesicles, lysosomes, peroxisomes 5. Cytoskeletal components and localization of organelles 6. How to prove co-localization of ER or GA with the microtubules