Histology II Flashcards
- What are the 3 types of epithelial tissues?
Contiguous cells, mucosa membrane and serous membrane
- Definition of Histology
Histology is the study of the microscopic structure of biological material and the ways in which individual components are structurally and functionally related.
- What are the characteristics of epithelium cells?
- A vascular
- Closely packed
- Have basal lamina
- What are the epithelium tissues derived from Ectoderm?
Skin, oral mucosa, glands of skin, mammary glands
- What are the epithelium tissues derived from Endoderm?
Liver, pancreas, respiratory and gastrointestinal tracts
- What are the epithelium tissues derived from Endoderm?
Liver, pancreas, respiratory and gastrointestinal tracts
- Definition of Tissue and the four main types
Tissue: collections of cells having similar morphological characteristics The four main types are: Epithelial Tissues Connective Tissue Muscular Tissues Nervous Tissues
- What composes the basement membrane?
Basal lamina -> lamina lucida + lamina densa
Lamina Retricularis
- What composes the lamina lucida?
Laminin, entactin
Laminin Receptors: integrins + dystroglcans
- What composes the lamina densa?
Collagen type 4 - anchoring
Collagen type 7 - fibrils
- What composes the lamina retricularis?
Fibroblasts, collagen 1+3
- Where can we find simple cuboidal epithelium?
Ducts of glands, Ovary covering, Kidney tubules
- Tissue preparation for TEM
- Fixation: glutaraldehyde and osmium tetraoxide
- Dehydrate and infiltration with some specific metal tissues.
- Embed and block fixed tissues in plastic.
- Cut into ultra-thin slices (50 nm thick)
- Stain sections with heavy metal salts (lead citrate and uranyl acetate) that bind nucleic acids & proteins.
- Where can we find stratified squamous epithelium?
Lining of mouth, oral pharynx, Vagina, Skin + keratinized
- Where can we find simple columnar epithelium?
Digestive tract, gall bladder, ducts
- Light microscopy description.
The light of the microscope is projected through the mirror to the condenser lens and to the specimen.
Objective lens magnifies the image of specimen (can be 4X, 10X, 40X to 100X).
The ocular lens projects to the eyes (10X).
- Magnification of Light Microscopy (formula)
Magnification = Ocular Lens x Objective Lens
- Resolution of Light microscopy
Anything smaller than 0.2 μm is out of resolution. (e.g. cellular membrane)
- Tissue preparation for light microscopy
- Stabilize cellular structures by chemical fixation.
- Dehydrate and infiltrate tissues with paraffin or plastic.
- Embed fixed tissue inside paraffin or plastic box.
- Cut into thin slices of 3-10 micrometer thick.
- Rehydrate and stain with Hematoxylin.
- Counter-stain with Eosin.
- Description of TEM (Transmission electron microscope)
The source of electrons is the cathode.
The electrons pass through the specimen and hit the bottom of the microscope, where the image viewing screen is.
- Alcian blue / Periodic acid-Schiff (PAS) Staining
- Strongly acidic mucins : blue
- Nuclei: pink, red
- Cytoplasm: pale pink
PAS detects polysaccharides as well as neutral and acidic mucins, allowing identification of hepatic glycogen, intestinal goblet cells and basal laminae.
- Where can we find trasitional epithelium?
Urinary system
- Where can we find Columnar pseudo-stratified epithelium?
Respiratory airways.
Usually contain cilia.
- What are the 3 cell surface specializations?
Brush border/microvilli, streocilia, cilia
- What composes Microvilli?
Actin, villin, fibrin, connected to the terminal web (intermediate filament) which is composed by spectin and actin
- Direct Immunofluorescence microscopy
Fluorophore is directly bound to antigen.
Fluorophore is the molecule that gives color to the signal.
- Indirect Immunofluorescence microscopy
Primary antibody recognizes the antigen and after that secondary antibody (which is marked with fluorophore) is bound to primary antibody.
- Confocal Microscopy
Resolution: 0.2 μm
Bigger clarity than light microscope.
Confocal microscopy can be used when there are 3D organs.
Specific markers can be used in order to identify specific molecules (Sox9 for stem cells).
- Golgis’ “black reaction” Staining method
- Fixation of nervous tissue blocks in potassium dichromate (2-2.5%)
- Takes 1 to 50 days or even longer
- Immersion in silver nitrate for silver chromate precipitation to fully impregnating cells in the nervous tissue
Turns most of the nervous tissue into black fibers
- Red Oil Staining
Used to demonstrate the presence of fats or lipids in fresh frozen tissue.
Performed on fresh frozen sections because fixatives containing alcohols, or routine tissue processing, will remove lipids.
- The Picro-Sirius Red Stain
Specific for collagen I and III fibers
Used in paraffin embedded tissue sections
- Haematoxylin and Eosin Staining
Haematoxylin: -Binds to basophilic substances (DNA) -Stains nuclei in blue-violet Eosin: -Binds to acidophilic substances (most proteins) -Stains cytoplasm in red or pink
- Alcian blue / Periodic acid-Schiff (PAS) Staining
Strongly acidic mucins → blue
Nuclei → pink to red
Cytoplasm → plae pink
PAS detects polysaccharides as well as neutral and acidic mucins.
- Types of cutting
Cross sectional (transverse)
Longitudinal (Horizontal)
Sagittal (Vertical)
- Functions of ECM
- Modify the morphology and functions
- Modulate survival of cells
- Regulate the migration of cells
- Direct mitotic activity
- Form functional associations with cell
- Cell Differentiation
The process by which unspecialized cells become specialized, in order to carry out specific functions.
- Stem cells definition
Cells that can proliferate without limit and can
differentiate into specialized cells.
- The three types of stem cells
Totipotent
Pluripotent
Multipotent
- Totipotent stem cells characteristics
- Arise directly from the division of the zygote.
- Can differentiate into any type of cell.
- Present in the blastocyst.
- Pluripotent stem cells characteristics
- Precursors to the fundamental tissue layers
2. Can differentiate into only one type of human tissue (epithelial, connective, muscle and nervous).
- Multipotent stem cells characteristics
- Can differentiate into different cell types in a given cell lineage.
We can say that the cells that derive from the same multipotent stem cell have the same progenitor.
- ON/OFF mechanism
In a certain cell, only some genes are expressed, while other are silent. This leads to cell differentiation.
It’s regulated by trascription factors.
- Definition of Induced Pluripotent Stem Cell (iPS)
A cell that has been reprogrammed, through TFs, to act like a pluripotent stem cell.
- Description of Yamanaka’s experiments (iPS)
- 24 possible TFs involved.
- He proved that when all of them acted
together on the DNA of a differentiated cell they were capable of taking it back to pluripotency, generating iPS. - He tested each of the 24 TFs one to see if there was a specific one responsible for this process.
- Four TFs necessary: Oct ¾, Sox2, Klf4, c-Myc.
- He had to prove that the cells he obtained were actually stem cells.
- Applications of iPS
- Regenerative medicine (in vivo)
- Toxicology (in vitro)
- Disease model, drug screening (in vitro)
- What are the three germ layers?
Ectoderm (outer)
Mesoderm (middle)
Endoderm (inner)
- Which tissues originate from the Ectoderm?
- Epidermis
- Glands on the skin
- Mouth between cheek and gums
- Some cranial bones
- Pituitary and adrenal medulla
- Nervous system
- Anus
- Which tissues originate from the Mesoderm?
- Connective tissues (proper, bones, cartilage, blood) 2. Endothelium of blood vessels
- Muscles
- Synovial and serous membranes
- Lining of body cavities and gonads
- Kidneys
- Which tissues originate from the Endoderm?
- Epithelium in the lungs
2. Part of the digestive system and part of its glands.
- Cell membrane functions
- Maintaining the structural integrity of the cell
- Selective permeability
- Regulating cell-cell interaction, recognition and communication between cells
- Interface between the cytoplasm and the external environment
- Establishing transport systems for specific molecules
- Transducing extracellular physical or chemical signals into intracellular events
- Endocytosis definition
It’s the mechanism by which the cell lets substances
inside the cell
- Phagocytosis definition
It’s a type of endocytosis that uses phagosomes, large vesicles (>250 nm).
For bigger molecules.
- Pinocytosis definition
For smaller molecules, uses vescicles of <150 nm.
These vescicles must be coated by clathrin.
- What happenes to pinocytic vescicles after their formation?
They lose the clathrin coat and fuse with endosomes.
Their content can be either degraded through lysosomes, or recycled through endosomes.
- What factor mediates pinocytosis?
EGF (epidermal growth factor)
- Cytoskeleton functions
- Shapes the cell’s form
- Controls the migration of the cells (ability of the
cells to move) - Forms an internal pathway that allows the movement of organelles within the cell.
- What are the three components of the cytoskeleton?
- Microtubules
- Thin filaments (or microfilaments)
- Intermediate filaments
- What’s in common with sweat glands, sebaceous glands and hair follicles?
Their are extended from the epidermis to the dermis
- What epigenetic regulations affect the skin?
DNM1 (DNA methyl transferase) - found more in the undifferentiated basal
Histone H3 Lysine27 trimethylation
Histone acetylation
All the processes decrease during the differentiation
- Where can we find microvilli/brush border?
Intestine, kidney proximal tubule cells
- What is the Treadmilling process in thin filaments?
- G-Actin-ATP associates with the PLUS END, where ATP is hydrolyzed to ADP.
- ADP-actin dissociates from filaments more readily than ATP-actin, so the plus end will grow faster and the minus end monomers will dissociate faster.
- This results in a net gain on the plus end, and a net loss on the minus end.
- Once a cell reaches the right size of the filament, it will put a capping protein on the plus
end, stopping the growing process.
- What composes Stereocilia?
Actin filaments, not motile
- What composes Cilia?
Anexome, made by 9 + 2 microtubules and dynein
- Where can we find cilia?
In respiratory system, oviduct
- What are the types of junctions?
Zonulae occludentes
Zonulae Adherents
Desmosomes
Hemidesmosomes
- What composes Zonulae Occludentes?
Claudius (e-cadherins), occludins, nestins, JAM
Proteins named ZO1->4
- What composes Zonulae Adherentes?
Outside: Cadheins (Ca dependent transmembrane linker)
Inside:
Vinculin, Catenin, alpha-actinin
Space diameter: 15-20 nm
- What composes Desmosomes?
Desmocollins, desmogleins connected to cadherins
Plano globing, plakophilins, held together by desmoplakin + keratin
Space diameter: 30 nm
- Where can you find compound acinar exocrine glands?
Parotid gland
Pancreas
- What are the types of secretion?
Serous, mucous, mixed
- Where can you find serous secretion?
Sweat, milk, tears, digestive juice from pancreas
- Where can you find mucous secretion?
Salivary glands + ??
- Where can you find mixed, mucous + serous secretion?
Salivary glands
- What are the methods of secretion?
Merocrine - exocytosis
Apocrine - small portion of the apical cytoplasm
Holocrine - the mature cell dies and become the secretory product
- Describe sweat glands
Simple coiled tubular
Merocrine/apocrine
Serous
- Describe Sebaceous glands
Simple branched acinar
Holocrine
Make sebum
Androgens stimulate it
If blocked + inflammation can cause acne
- Describe prostate gland
Tubuloalveolar gland lined with epithelium
Concretions can be in the center - calcified glycoproteins
Serous - citric acid,lipids, proteolytic enzymes, phosphotase, fibrinolysin
Regulated by dihydrotestosterone
- Describe salivary glands
compound tubulo/acinar
Serous from acinus
Mucous from tubular
Secrete lysozyme, lactoferrin, IgA
Covered with mayo epithelial cells
Ducts intercalated to very large principal ducts
3 glands:
- parotid: biggest, 30% of saliva, only serous
- submandibular: 60% of saliva, 95% serous 5% mucous
- sublingual: smallest, mostly mucous tubular units, coupled with Demilumes - serous cells
- What is it Metaplasia?
Changing of the type of the epithelium, pseudostratified ciliated ep. -> squamous in a smoker’s lungs (Camilla quit smoking!)
- What is the name of tumors of the epithelium?
Carcinoma/adenocarcynoma
- Describe the liver
Hepatocyte hexagon shape, classical lobes
Each lobe has single central vein
In lateral domains- bile canlicul
In sinusoidal domain-space of Disse, microvilli, lipid droplets, glycogen
Surrounded with Glisson’s capsule connective tissue- conduct of blood vessels, lymph ves., bile duct
Blood - 25% hepatic artery 75% portal vein
Bile leaves through porta hepatic
- What is the rete apparatus?
Dermal ridges (papillae) that are the interface of the epidermis and the dermis
- How is the interface of the epidermis and the dermis called?
Dermal ridges (papillae)/rete apparatus
- What is the diameter of the skin?
Thin - 0.07-0.12 micro m
Thick - 0.8 - 1.4 micro m
- What are the names of the layers of the skin?
Stratum corneum Stratum lucidum Stratum granulosum Stratum spirosum Stratum basale
- What is the name of the cells that compose the skin?
keratinocytes
- Which other cells are found in the skin?
- Langerhan cells - antigen presentation
- Merkel cells - light touch sensations mechanoreceptors
- Melanocytes
- What are the two layers of the dermis?
Papillary - form dermal ridges with epidermis, loose
Retricular layer - thick, deeper denser
- What’s the papillary layer made of?
In dermis - made of collagen 3 + elastic fibers - loose
- What’s the retricular layer made of?
Type 1 collagen - thick
- What is the superficial fascia?
Hypodermis, binded to dermis
- What’s in common with sweat glands, sebaceous glands and hair follicles?
Their are extended from the epidermis to the dermis
- What epigenetic regulations affect the skin?
DNM1 (DNA methyl tranferase) - found more in the undifferentiated basal
Histone H3 Lysine27 trimethylation
Histone acetylation
All the processes decrease during the differentiation
- Define the Plus end and the Minus end in thin filaments.
The plus end is the rapidly growing filament.
The minus end is the slowly growing filament.
The enlargment is through the Plus side.
- What is the Treadmilling process in thin filaments?
G-Actin-ATP associates with the PLUS END, where ATP is hydrolyzed to ADP.
ADP-actin dissociates from filaments more readily than ATP-actin, so the plus end will grow faster and the minus end monomers will dissociate faster.
This will result in a net gain in the plus end, and a net loss in the minus end.
Once a cell reaches the right size of the filament, it will put a capping protein on the plus
end, thus stopping the growing process.
- What is the major capping protein?
Gelsolin, once attached to the plus
end, terminates the elongation of microfilaments.
- What promote the elongation of thin filaments?
Poly-phospho-inositide, it removes the gelsolin cap, permitting elongation of the actin filament.
- What are the three types of G-actin?
- α-actin; MUSCLES
- β-actin and γ-actin : any other cell
- What proteins associate with actin?
Actin is responsible for motility via association with different actin binding protein.
The most common is myosin. Others are spectrin, fimbrin, gelsolin and talin.
- What are the 4 thin filaments arrangements?
- The Contractile Bundle (stress fibers)
- Gel-like network, elements of the cell cortex
- Filopodium, made of tight parallel bundle
- Dendritic network
- Contractile Bundle roles and characteristics
- Are associated with myosin
- The actin filaments are arranged loosely, parallel to each other, with plus and minus end alternating in direction
- Role: organelles and vesicles movements within the cells
- Endo and Exocytosis
- Gel-like network role and characteristics
- Provides the structural foundation of the cell cortex
- Contrains filamin, that assists in the formation of the actin network and gives stiffness to the network
- Parallele bundles role and characteristics
2 proteins: Fimbrin (in tiny spikes) and Villlin
- Fimbrin allows movement
- VIllin forms the microvilli
- How are cells able to move?
Thanks to the lamellopodia: this extracellular protrusion
enables cancer cell to invade the surrounding tissues.
When we remove one of the proteins that binds to the filaments, the cells can’t move and we’ll have filopodia.
- What is the focal region?
It’s a zone of connection between the cell and the extracellular matrix.
It’s an anchoring junction of the cell to a non-cellular substrate.
- How do cells mantain attachment?
Bundles of the actin cytoskeleton attach to the transmembrane linkers integrins, that in turn bind glycoproteins such as fibronectin in the ECM, permitting attachment.