Professional Documents
Culture Documents
Histology:
branch of anatomy study of tissues does not deal only with tissues but also with functions
3 Tissue Preparation
y
Steps in Tissue Preparation Fixation Dehydration Clearing Embedding Sectioning Mounting Staining
4 Fixation
y
Fixation
refers to the treatment of tissue with chemical agents that retards the alteration of the tissue subsequent to death or removal from the body maintains normal structure of the tissue most common fixatives used in light microscopy are: Both of these fixatives cross-link proteins, thus maintaining a life-like image of the object. neutral buffered formalin Bouins fluid
5 Dehydration
y
Dehydration
process of removing water from the tissues graded series of alcohol baths beginning with 50% to 100% alcohol are
Clearing
process of treating the tissues with xylene xylene is a chemical that is miscible with melted paraffin the tissue becomes transparent with xylene
7 Embedding
y
Embedding done to distinguish the overlapping cells in a tissue and extracellular matrix from one another usual embedding medium is melted paraffin the tissue is placed in a suitable container of melted paraffin until the tissue is completely infiltrated
Trimming
removal of excess paraffin wax from the tissue block cutting the tissue with the use of a microtome ideal thickness of tissue is about 5 to 10 um Can also be performed on specimens frozen in liquid nitrogen or on the rapid freeze bar of a cryostat
Sectioning
Mounting
placing tissue sections on adhesive-coated glass slide provides contrast to the tissue with the use of stains stains that are used are grouped into 3 classes: stains that differentiate between acidic and basic components of the cell specialized stains that differentiate the fibrous components of the extracellular matrix
Staining
10 Staining
y
Most commonly used stain in histology: Hematoxylin and Eosin (H & E) Hematoxylin is a base that colors the acidic components of the cells a bluish tint DNA and RNA, nucleus and cytoplasm rich in ribosomes have acidic pH and stain dark blue (basophilic) Eosin is an acid that dyes the basic components of the cell a pinkish color regions of the cytoplasm have basic pH and stain pink (acidophilic)
11 Light Microscope
y
permits a high magnification and good resolution of being viewed several types of light microscopes are distinguished by: type of light used as a light source manner in which they use the light source
Digital Imaging Technique utilize computer technology to capture and manipulate histologic techniques images are stored in digital format and therefore are easy to archived and retrieval is almost instantaneous and thus allows electronic transmission of images advantages immediate visualization of acquired image digital modification of the image capability of enhancing the image by the use of commercially
available software
Histochemistry method of staining provides information concerning the presence and location of intracellular and extracellular macromolecules tissue structures are colored when a chemical group (e.g. carboxyl, phosphoric or aldehyde) reacts with the stain suitable for diagnosis of pathologic conditions not suitable for routine tissue staining
Histochemistry capitalize on enzyme activity, chemical reactivity or other physicochemical phenomena associated with the constituent of interest permits relatively good localization of some enzymes and macromolecules reactions of interest are monitored by the formation of an insoluble precipitate that takes on a certain color performed on frozen tissues and can be applied to both light and electron microscopy Ex. Periodic Acid Schiff Stain
Immunocytochemistry uses fluoresceinated antibodies and antiantinbodies to provide more precise intracellular and extracellular localization of macromolecules than is possible with histochemistry
there are two methods of antibody labeling: Direct: antibody against the macromolecule is labeled with a fluorescent dye; antibody is made to react with the macromolecule and the resultant complex is viewed with a fluorescent microscope
Indirect: fluorescent-labeled antibodies are prepared against an antibody that reacts with a particular antigen ; when viewed with a fluorescent microscope, the region of fluorescence represents the location of the antibody.
Autoradiography uses the incorporation of radioactive isotopes into macromolecules, which are then visualized by the use of an overlay of film emulsion
Autoradiography
20 Electron Microscopy
y
makes use of electron beams instead of light rays and magnetic fields instead of lenses 2 types:
requires heavy metal precipitation techniques Fixatives used include buffered solutions of glutaraldehyde, paraformaldehyde, osmium tetroxide and potassium permanganate ( these does not only preserve fine structural details but also act as electron dense
used to view the surface of a solid specimen specimen is prepared in a special manner that permits a heavy metal such as gold or palladium, to be deposited on the specimens surface
Freeze-Fracture Technique reveals internal aspects of membranes tissue does not suffer mechanical damage quick-frozen specimens are treated with cryopreservatives that do not develop ice crystals during the freezing process replica of the surface is generated as the specimen is hit by a supercooled razor blade that fractures along cleavage planes, which are regions of least molecular bonding fracture face is coated at an angle by evaporated platinum and carbon, forming accumulations of platinum on one side and no accumulation on
the opposite side next to the projection, thus generating a replica the replica is then examined by transmission electron microscopy
Freeze-Fracture Technique
All cells possess certain unifying characteristics and thus can be described in general terms:
4 Introduction
o o
Cells are the basic functional units of complex organisms. The human body is composed of more than 200 different types of cells, each performing a different function
5 Introduction
y
Protoplasm, the living substance of the cell is divided into two compartments:
6 Cell Membrane: A Selectively Permeable Barrier The cell membrane forms a selectively permeable barrier between the cytoplasm and the extracellular fluid
Functions
o o
o o o
maintains the structural integrity of the cell controls movements of substances in and out of the cell (selective permeability) regulates cell to cell interactions recognition via receptors, antigens, foreign cells and altered cells acts as an interface between the cytoplasm and the extracellular fluid
Cell Membrane The cross section of the cell membrane shows two different structures:
o o
The phospholipids are the round yellow structures with the blue tails. The proteins are the lumpy structures that are scattered among the phospholipids.
This is a representation of a phospholipid The yellow structure represents the polar head, which is the hydrophillic or water loving section of the phospholipid. The blue tails that come off the sphere represent the non-polar tails which are the hydrophobic or water fearing ends of the phospholipid.
The two long chains coming off the bottom of this molecule are made up of carbon and hydrogen. These chains are not attracted to water. These are the non-polar tails. At the other end of the phospholipid is a phosphate group and several double bonded oxygens. The atoms at this end of the molecule are not shared equally. This end of the molecule has a charge and is attracted to water. This is the polar head. If you mix phospholipids in water they will form these double layered structures. The hydrophillic ends will be in contact with water. The hydrophobic ends will face inwards touching each other.
Floating around in the cell membrane are different kinds of proteins. These are generally globular proteins. They are not held in any fixed pattern but instead float around in the phospholipid layer. Generally these proteins structurally fall into three catagories:
They are multipass membrane transport proteins that possess binding sites or molecules on both sides of the lipid bilayer. Transport may be passive, along an electrochemical concentration gradient; or active against a gradient.
They do not extend through the membrane and they bond and drag molecules through the bilipid layer and release them on the opposite side.
Steroids are sometimes a component of cell membranes in the form of cholesterol. When it is present, it reduces the fluidity of the membrane. Not all membranes contain cholesterol.
o o
The cell membrane's function, in general, revolves around its membrane proteins. General classification of membrane proteins include: receptor proteins which allow cells to communicate transport proteins which regulate what enters or leaves the cell marker proteins which identify the cell
These proteins are used in intercellular communication. This causes the receptor protein to release a signal to perform some action.
electrochemical gradient or utilize ATP-driven transport mechanisms to ferry specific substances across the cell membrane against the concentration gradient 17 Cell Membrane Function: Membrane Transport Proteins
y
Transport may be coupled, two different molecules moving in the same direction Symports also use the process of diffusion. In this case, a molecule that is moving naturally into the cell through diffusion is used to drag another molecule into the cell. In this example glucose hitches a ride with sodium
o o
Symport coupled transport is the simultaneous transport of two different molecules or ions across a lipid bilayer membrane in the same direction.
y y
Antiport coupled transport involves two different molecules or ions across a lipid bilayer membrane in opposite directions.
Marker proteins extend across the cell membrane and serve to identify the cell. They are as unique as fingerprints. They play an important role in organ transplants.
o o
Channel proteins are also membrane transport proteins that may be gated or ungated They are incapable of transporting substances against a concentration gradient. Channel proteins extend through the bilipid layer. They form a pore through the membrane that can move molecules in several ways. In some cases the channel proteins simply act as a passive pore. Molecules will randomly move through the opening in a process called diffusion. This requires no energy, molecules move from an area of high concentration to an area of low concentration.
Some proteins actively use energy from the ATPs in the cell to drag molecules from areas of low concentration to areas of high concentration (working directly against the concentration gradient). The sodium/potassium pump is an example in which the energy of a phosphate is used to exchange sodium ions for potassium ions.
24 Cytoplasm The term cytoplasm refers to everything between the cell membrane and the nuclear envelope. It consists primarily of water and contains various organelles as well as salts, dissolved gases and nutrients.
Golgi Apparatus
Functions in synthesis of carbohydrates and in the modification of and sorting of proteins manufactured in the rough endoplasmic reticulum
Lysosomes
o o
Not studded with ribosomes For carbohydrate and lipid synthesis as well as detoxification
Studded with ribosomes Vesicles associated with the RER and golgi possess a protein coat and surface markers
Mitochondrion
Possess their own DNA and performs oxidative phosphorylation and lipid synthesis. Note that the inner membrane is thrown into folds called cristae.
Centrioles
o
These are small cylindrical structures consisting of nine microtubule triplets. They constitute the core of the microtubule organizing center or the centrosome.
Peroxisomes
Ribosomes o They are small particles that provide cell surface for protein synthesis.
o
Each ribosome is composed of a small subunit (40S) and a large subunit (60S), both of which are manufactured or assembled in the nucleolus and released separately in the cytosol. The 40S subunit is composed of 33 proteins and an 18S rRNA. The 60S subunit has 49 proteins and 3 rRNAs.
o o o
33 Nucleus
y
Nucleus
o o
This is the largest organelle of the cell. It contains nearly all the DNAs possessed by the cell and all the mechanisms for RNA synthesis. It is surrounded by a nuclear envelope which is composed of two parallel unit membranes that fuse with each other at certain regions to form perforations known as nuclear pores. It houses three major components chromatin nucleolus nucleoplasm
34 Nucleus
y
Chromatin
Chromatin is the complex of DNA and protein found inside the nuclei of eukaryotic cells The nucleic acids are in the form of double-stranded DNA. The major proteins involved in chromatin are histone proteins. The functions of chromatin are: to package DNA into a smaller volume to fit in the cell to strengthen the DNA to allow mitosis and meiosis and to serve as a mechanism to control expression
o o o
35 Nucleus
y
Nucleolus
This is a deeply staining non-membrane bounded structure within the nucleus that is involved in the rRNA synthesis and in the assembly of small and large ribosomal subunits
Slide Show Outline 1 CELL CYCLE and REPLICATION 2 The Cell Cycle 3 Mitosis
y
y y y
Mitosis o When a eukaryotic cell divides into two, each daughter or progeny cell must receive a complete set of genes (for diploid cells, this means 2 complete set of genomes=2n)
o
In human cells there are 23 pairs of chromosomes, 22 pairs are autosomes and 1 pair constitute the sex chromosomes (XX for
4 Mitosis
y y y
Mitosis is preceded by an exact duplication of the DNA and associated proteins in the nucleus This occurs during interphasethe period between actual mitotic divisions. After this doubling, the chromosomes are organized, complete division and are then equally distributed to the two daughter cells during mitosis.
5 Mitosis
y y
All somatic cells pass through the mitotic and interphase periods at one time or another. The duration of the mitotic period is usually about 0.5 to 2 hours, while the intermitotic period can vary from a few hours to many years.
6 Mitosis
y
The mitotic index (proportion of cells undergoing division at any one time) varies greatly depending upon the type of tissue and the physiological needs of the organism. A high mitotic index is observed in: o all growing embryonic tissues o certain adult tissues such as : bone marrow intestinal crypts cancer cells
7 Interphase
y
Interphase o Interphase is not a stage of mitosis o The chromosomes cannot be distinguished and appear as scattered granules connected by a network of pale-staining strands within a distinct nuclear membrane. o A nucleolus is usually present
8 Stages of Mitosis
Mitosis is divided into 4 stages for convenience of study: o Prophase o Metaphase o Anaphase o Telophase
9 Interphase
y
Interphase o is not a stage of mitosis o Longest period in the cell cycle o Divided into 3 stages called into order: G1 phase S phase G2 phase o The chromosomes cannot be distinguished and appear as scattered granules connected by a network of pale-staining strands within a distinct nuclear membrane. o A nucleolus is usually present
10 Prophase
y
Prophase o Chromatin materials which are made up of DNA and associated proteins condense into chromosomes o Nucleolus fades o Chromosomes split and form 2 sister chromatids o The 2 sister chromatids remain attach at a region called centromere
11 Prometaphase
y
Prometaphase
o o
Dissolution of nuclear envelope Spindle fibers come in contact with the chromosomes
12 Metaphase
y
Metaphase o chromosomes become arranged at the equatorial plate which is midway between the two pairs of centrioles
the spindle is now fully formed and does not stain (achromatic) and consists of fine microtubules, some of which are attached to the chromosomes. During this period the centromeres begin to show indications of splitting
13 Anaphase
y
Anaphase o each chromosome pair completes its splitting, and the two daughter chromosomes move toward opposite poles o spindle microtubules may be seen between the retreating chromosomes o spindle fibers shorten through a process known as depolymerization (removal of tubulin proteins)
14 Telophase
y
Telophase o telophase, the chromosomes have reached the spindle poles and appear as a dense, basophilic mass within which individual chromosomes cannot be defined. o the nuclear membrane reappears, and the outlines of the chromosomes disappear leaving scattered chromatin granules connected by a pale-staining network. o nucleoli reappear and seem to be associated with a particular chromosome.
15 Cytokinesis
y
Cytokinesis o the division of the cytoplasm, usually occurs during the telophase, but the synchrony between nuclear and cytoplasmic telophase, is not constant o cytokinesis may begin as early as late anaphase or be delayed beyond the nuclear reconstruction of telophase
Cells in mitosis are easy to spot. Instead of a nucleus, the chromosomes are visible as tangled, dark-staining threads.
y y y
We call these "mitotic figures." Counting mitotic figures sometimes helps the pathologist tell benign from malignant. If you see a nuclear membrane, it is not a mitotic figure
17 Meiosis
y
Meiosis is the type of cell division by which germ cells (eggs and sperm) are produced. Meiosis involves a reduction in the amount of genetic material Meiosis comprises two successive nuclear divisions with only one round of DNA replication. Four stages can be described for each nuclear division Interphase: Before meiosis begins, genetic material is duplicated
y y y
2n
Prophase 1 o Duplicated chromatin condenses. o Each chromosome consists of two, closely associated sister chromatids o Crossing-over can occur during the latter part of this stage Metaphase 1 o Homologous chromosomes align at the equatorial plate
Telophase 1 o Two daughter cells are formed with each daughter containing only one chromosome of the homologous pair.
y y y y
Second division of meiosis: n n o Gamete formation Prophase II o chromosomes shorten, condense and thicken Metaphase II o Chromosomes align at the equatorial plate Anaphase II o Centromeres divide and sister chromatids migrate separately towards opposite pole Telophase II o Cell division is complete. o Four haploid daughter cells are formed
20 Gametogenesis
y y
Meiosis is studied in relation to gametogenesis Gametogenesis is the general process of gamete formation in both males and females. Female gametogenesis: oogenesis Male gametogenesis: spermatogenesis
y y
21 Oogenesis
y
Oogenesis is the process of meiosis in female organisms from an oogonium to a primary oocyte, to a secondary oocyte, and then to an ovum. Oogenesis begins soon after fertilization, as primordial germ cells travel from the yolk sac to the gonads, where they begin to proliferate mitotically. They become oocytes once they enter the stages of meiosis several months after birth. Now called primordial follicles, they are made up of oogenic cells from the primordial germ cells surrounded by follicle cells from the somatic line.
y y
The oocyte is then arrested in the first meiotic prophase until puberty.
22 Oogenesis
y y y y
At puberty, between 4 to 10 follicles begin to develop, although only 1-2 are actually released. Surrounding each oocyte is a zona pellucida, membrana granulosa, and theca cell layer. Each oocyte finishes its first meiotic division, creating a secondary oocyte and polar body, which serves no further function. It begins the next meiosis cycle and is arrested in its second metaphase, at which point it is released from the ovary in ovulation. It will not finish the meiosis cycle until it is fertilized by a sperm.
23 Oogenesis 24 Spermatogenesis
y y
Spermatogenesis: the process by which stem cells develop into mature spermatozoa. There are three phases: o Spermatocytogenesis (Mitosis) o Meiosis o Spermiogenesis
25 Spermatogenesis
y
Spermatocytogenesis o Also called mitosis o Stem cells Type A spermatogonia divide mitotically to produce cells that begin differentiation into Type B spermatogonia Spermatogonia have spherical or oval nuclei, and rest on the basement membrane
26 Spermatogenesis 27 Spermatogenesis
y
Meiosis: o Cells in prophase of the first meiotic division are primary spermatocytes. o They are characterized by highly condensed chromosomes giving the nucleus a coarse chromatin pattern and an intermediate position
o o
in the seminiferous epithelium. This is a long stage, so many primary spermatocytes can be seen. Primary spermatocytes go through the first meiotic division and become secondary spermatocytes. The cells quickly proceed through this stage and complete the second meiotic division. Because this stage is short there are few secondary spermatocytes to be seen in sections. is the process by which the diploid number of chromosomes present in spermatogonia (the stem cells) is reduced to the haploid number present in mature spermatozoa
28 Spermatogenesis 29 Spermatogenesis
y
Meiosis o The products of the second meiotic division are called spermatids. o They are spherical cells with interphase nuclei, positioned high in the epithelium. o Since spermatids go through a metamorphosis into spermatozoa, they occur in early through late stages. o All of these progeny cells remain attached to each other by cytoplasmic bridges. o The bridges remain until sperm cells are fully differentiated
30 Spermatogenesis
y
Spermiogenesis o This is the metamorphosis of spherical spermatids into elongated spermatozoa. o No further mitosis or meiosis occurs. o During spermiogenesis, the acrosome forms, the flagellar apparatus forms, and most excess cytoplasm (the residual body) is separated and left in the Sertoli cell. o Spermatozoa are released into the lumen of the seminiferous tubule. o A small amount of excess cytoplasm (the cytoplasmic droplet) is shed later in the epididymis.
31 Spermatogenesis 32 Spermatogenesis
Spermiogenesis
a process of metamorphosis from a round cell with typical organelles to a highly specialized, elongated cell well adapted for traversing the male and female reproductive tracts and achieving fertilization of an egg
Cytogenetics is the study of chromosomes and the related diseased states caused by numerical and structural chromosome abnormalities. A variety of cell or tissue types can be used to perform these studies.
3GENE
4GENOME
y
5GENE POOL
y
6HOMOLOGOUS CHROMOSOMES
y
Two physically identical chromosomes with the same gene loci but not necessarily the same alleles; one is of maternal origin and the other paternal
7SEX CHROMOSOMES
y
8AUTOSOMES
y
9GENE LOCUS
y
1ALLELE 0 y Any member of a given pair of gene; any alternative form that a particular gene can take 1GENOTYPE 1 y The genetic make-up of an individual; alleles that a person has for a particular trait 1PHENOTYPE 2 y A detectable trait, such as eye color, blood type, appearance of fingers, etc.
1RECESSIVE ALLELE 3 y An allele that is hidden/not expressed in the presence of a dominant gene 1DOMINANT ALLELE 4 y Allele that is always expressed whenever present 1HOMOZYGOUS 5 y Pure individual; with identical alleles for a given gene 1HETEROZYGOTE 6 y An individual with two different alleles for a given gene 1CARRIER 7 y A person who carries a recessive allele but does not phenotypically express it 1CODOMINANCE 8 y A condition in which both alleles are fully expressed when present in the same individual 1INCOMPLETE DOMINANCE 9 y A condition in which two alleles are both expressed when present in the same individual and the phenotype is intermediate between those which each allele would produce alone 2PLEIOTROPY 0 y A condition in which a single phenotype results from the combined action of genes at two or more different loci, as in eye color 2SEX LINKAGE 1 y Inheritance of a gene on the x or y chromosome so that the associated phenotype is expressed more in one sex than in the other
2PENETRANCE 2 y The percentage of individuals with a given phenotype who actually exhibit the phenotype predicted from it 2CHROMOSOMES 3 y Three dimensional structure in the cell that is chemically made up of DNA 2G BANDING CHROMOSOMES 4 y Characteristic banding patterns caused by staining the chromosomes with dye called Giemsa 2FISH 5 y A molecular cytogenetic technique in which fluorescent gene probes are used to determine the presence or absence of chromosomes, DNA specific sequences or genes 2M-FISH 6 y Multi-Color Fluorescent in situ hybridization. y All chromosomes are distinguished by a specific color 2CGH 7 y Comparative Genomic Hybridization Comparative genomic hybridization (CGH) is a molecular-cytogenetic method for the analysis of copy number changes (gains /losses) in the DNA content of tumor cells. The method is based on the hybridization of fluorescently labeled tumor DNA (frequently Fluorescein - FITC) and normal DNA (frequently Rhodamine or Texas Red) to normal human metaphase preparations. 2HISTONE 8 y http://images.search.yahoo.com/search/images/view?back=http%3A%2F%2 Fimages.search.yahoo.com%2Fsearch%2Fimages%3Fp%3DHistones%26ei %3DUTF-8%26fr%3Dyfp-t482%26fp_ip%3DPH%26x%3Dwrt&w=417&h=986&imgurl=www.unc.ed u%2Fdepts%2Fmarzluff%2Fhistone.jpg&rurl=http%3A%2F%2Fwww.unc. edu%2Fdepts%2Fmarzluff%2Fhistone.html&size=52.9kB&name=histone.j
pg&p=Histones&type=jpeg&no=2&tt=356&oid=e0e737270c654730&ei=U TF-8 2HISTONE 9 y Histones o combine with DNA to form nucleosomes, the fundamental structural units of chromatin. o Nucleosomes pack DNA in a stable coiled form in eukaryotic nuclei. o The total DNA complement of the nucleus, with associated histone and non-histone proteins, is broken into individual lengths; these are the chromosomes.
y
3NUCLEOSOME 0 y The coiling of DNA around nucleosomes and the further winding of nucleosomes into chromatin fibers greatly compact the DNA of the eukaryotic nuclei. y Winding the DNA into nucleosomes and chromatin fibers is estimated to shorten its length by at least a factor of 10,000. y Because packing of nucleosomes in chromatin fibers probably prevents access to DNA by the enzymes of transcription and replication, the fibers probably unwind for these activities to take place. 3NUCLEOSOME 1 y The coiling of DNA around nucleosomes and the further winding of nucleosomes into chromatin fibers greatly compact the DNA of the eukaryotic nuclei.
y
Winding the DNA into nucleosomes and chromatin fibers is estimated to shorten its length by at least a factor of 10,000. Because packing of nucleosomes in chromatin fibers probably prevents access to DNA by the enzymes of transcription and replication, the fibers probably unwind for these activities to take place.
3
y
3MITOSIS 4 y Retrieved from: http://images.search.yahoo.com/search/images/view?back=http%3A%2F%2 Fimages.search.yahoo.com%2Fsearch%2Fimages%3Fp%3DMITOSIS%26e i%3DUTF-8%26fr%3Dyfp-t482%26fp_ip%3DPH%26x%3Dwrt&w=500&h=600&imgurl=www.dartmo uth.edu%2F%7Ecbbc%2Fcourses%2Fbio4%2Fbio4lectures%2Fimages%2Fmitosis.JPG&rurl=http%3A%2F%2Fwarren.dusd.n et%2F%7Ecrobinson%2FCoordinated_Past_Web_Pages%2Fmitosis_meiosi s.htm&size=17.1kB&name=mitosis.JPG&p=MITOSIS&type=jpeg&no=12 &tt=15,737&oid=09d38de0a2885188&ei=UTF-8 3 TRISOMY 21 5 3TRISOMY 21 6 y Retrieved from:http://www.millerandlevine.com/genome/trisomy-21.jpg y http://www.woodbinehouse.com/images/large/down_syndrome_the_first_18 _months.jpg y http://www.down-syndrom.ch/Fotos/Welcom1.jpg 3CRI-DU CHAT 7 y http://images.search.yahoo.com/search/images/view?back=http%3A%2F%2 Fimages.search.yahoo.com%2Fsearch%2Fimages%3Fp%3DCRI%2BDU% 2BCHAT%2Bsyndrome%26ei%3DUTF-8%26fr%3Dyfp-t482%26fp_ip%3DPH%26x%3Dwrt&w=925&h=916&imgurl=www.busine ss.unet.com%2F%7Ecridchat%2Fgallery%2Fj%2Fjanel_1.jpg&rurl=http%3A% 2F%2Fwww.business.unet.com%2F%7Ecridchat%2Fgallery%2Fj%2Fjanel_1.htm&size=150.8kB &name=janel_1.jpg&p=CRI+DU+CHAT+syndrome&type=jpeg&no=2&tt =394&oid=6bea37bb9482898a&ei=UTF-8 3KARYOTYPE 8 y Retrieved from http://fig.cox.miami.edu/~cmallery/255/255hist/mcb10.0.karyotype.jpg
Organization of the chromosomes of an individual, lined up from largest to smallest according to the location of the centromere.
Slide Show
Cytogenetic analyses are almost always based on examination of chromosomes fixed during mitotic metaphase. During that phase of the cell cycle, DNA has been replicated and the chromatin is highly condensed. The two daughter DNAs are encased in chromosomal proteins forming sister chromatids, which are held together at their centromere. The centromere is the structure where the mitotic spindle attaches prior to segregation
3 Metaphase Chromosome
y
Metaphase chromosomes differ from one another in size and shape, and the absolute length of any one chromosome varies depending on the stage of mitosis in which it was fixed. However, the relative position of the centromere is constant, which means that that the ratio of the lengths of the two arms is constant for each chromosome. This ratio is an important parameter for chromosome identification.
4 Metaphase Chromosome
y
The ratio of lengths of the two arms allows classification of chromosomes into several basic morphologic types
5 Chromosomal Banding
y
Centromere position and arm ratios can assist in identifying specific pairs of chromosomes, but inevitably several or many pairs of chromosomes appear identical by these criteria. The ability to identify specific chromosomes with certainty was revolutionized by discovery that certain dyes would produce reproducible patterns of bands when used to stain chromosomes.
Chromosome banding has since become a standard and indispensible tool for cytogenetic analysis., and several banding techniques have been developed: o Q banding: chromosomes are stained with a fluorescent dye such as quinacrine o G banding: produced by staining with Giemsa after digesting the chromosomes with trypsin o C banding: chromosomes are treated with acid and base, then stained with Giemsa stain
6 Chromosome Banding
y
Each of these techniques produces a pattern of dark and light (or fluorescent versus non-fluorescent) bands along the length of the chromosomes. Importantly, each chromosome displays a unique banding pattern, analagous to a "bar code", which allows it to be reliably differentiated from other chromosomes of the same size and centromeric position.
7 Chromosome Banding
y
In the following figure, human chromosome pairs 1, 2 and 3 are seen with and without G banding.
Chromosomes are arranged into seven groups based on size and centromere location. The centromeres can be found in the: o Middle of the chromosome (median) o Near one end (acrocentric) o Between the middle and end: submedian
3 Group A Chromosomes
y y y
4 Group B Chromosomes
y y y
5 Group C Chromosomes
y y y
6 Group D Chromosomes
y y y
7 Group E Chromosomes
y y y
8 Group F Chromosomes
y
Chromosomes 19-20
y y
9 Group G Chromosomes
y y y
Cri du Chat Syndrome o deletion in chromosome no. 5 o defective larynx o mishapen ears o small head
y y
Klinefelter Syndrome o XXY o Phenotypically male Turner Syndrome o XO o Only viable human monosomy Metafemale XYY syndrome
Klinefelter syndrome (KS) is a disorder that occurs in some men who have more than one X chromosome (XXY). Virtually every cell in the body contains chromosomes, which carry the genes that determine many of our physical, intellectual, and emotional characteristics.
Males usually inherit a single X chromosome from their mother and a single Y chromosome from their father. Males with KS get at least one extra X chromosome from either their mother or their father. In most cases there is only one extra chromosome. The extra X chromosome is caused by a biological accident.
Normally, an egg has one X chromosome and a sperm has one X or one Y chromosome.
8 Sex Chromosomal Aberrations (Klinefelter Syndrome) Risk Factors for Klinefelter Syndrome A risk factor is something that increases your chances of getting a disease or condition. Women over age 35 have a slightly increased chance of having a child with KS. There are no other known risk factors for this disorder.
y y
9 Sex Chromosomal Aberrations (Klinefelter Syndrome) Although the chromosome variation XXY occurs in approximately 1 out of 1,000 live male births, many men with it do not develop KS. When KS does develop, it usually goes undetected until puberty and sometimes much later. It may present in childhood as learning problems, adolescence as excessive breast development, in adult life as infertility concerns and as an unexpected finding on an amniocentesis.
10 Sex Chromosomal Aberrations (Klinefelter Syndrome) Symptoms/Characteristics may include: o Tallness with extra long arms and legs o Abnormal body proportions (long legs, short trunk) o Enlarged breasts o Lack of facial and body hair o Small firm testes o Small penis o Lack of ability to produce sperm
o o o o
11 Sex Chromosomal Aberrations (Klinefelter Syndrome) 12 Sex Chromosomal Aberrations (Klinefelter Syndrome) 13 Sex Chromosomal Aberrations (Klinefelter Syndrome) 14 Sex Chromosomal Aberrations (Klinefelter Syndrome) 15 Sex Chromosomal Aberrations (Klinefelter Syndrome) 16 Sex Chromosomal Aberrations (Klinefelter Syndrome)
y
Treatment o Testosterone o When boys with KS are 10 to 12 years old, their hormone levels are checked yearly. o If their testosterone levels are low, then treatment with testosterone may be very helpful. o Men diagnosed in adulthood may also benefit from taking testosterone. o However, testosterone cannot reverse infertility.
Treatment Testosterone is most often given through regular shots in the form of depotestosterone. The benefits of this treatment include: o Increased strength o More muscular, male appearance o Growth of facial and body hair o Better self-esteem o Modulation of mood
o o o
18 Sex Chromosomal Aberrations (Turner Syndrome) 19 Sex Chromosomal Aberrations (Turner Syndrome) 20 Sex Chromosomal Aberrations (Turner Syndrome) 21 Sex Chromosomal Aberrations (Turner Syndrome) 22 Sex Chromosomal Aberrations (Turner Syndrome) 23 Sex Chromosomal Aberrations (Turner Syndrome) 24 Sex Chromosomal Aberrations (XYY Syndrome) 25 Sex Chromosomal Aberrations (XYY Syndrome)
Euploidy is the condition of having a normal number of structurally normal chromosomes. Euploid human females have 46 chromosomes (44 autosomes and two X chromosomes), and euploid bulls have 60 chromosomes (58 autosomes plus an X and a Y chromosome).
3 ANEUPLOIDY
Aneuploidy o is the condition of having less than or more than the normal diploid number of chromosomes, and is the most frequently observed type of cytogenetic abnormality. o abnormality in chromosome number o in other words, it is any deviation from euploidy, although many authors restrict use of this term to conditions in which only a small number of chromosomes are missing or added.
4 ANEUPLOIDY
y
Aneuploidy o is recognized as a small deviation from euploidy for the simple reason that major deviations are rarely compatible with survival, and such individuals usually die prenatally
5 ANEUPLOIDY
y
The two most commonly observed forms of aneuploidy are monosomy and trisomy: o Monosomy 2n-1 is lack of one of a pair of chromosomes an individual having only one chromosome 6 is said to have monosomy 6. A common monosomy seen in many species is X chromosome monosomy, also known as Turner's syndrome. XO is the only viable human monosomy Monosomy is most commonly lethal during prenatal development o Trisomy 2n + 1 is having three chromosomes of a particular type a common autosomal trisomy in humans is Down syndrome, or trisomy 21, in which a person has three instead of the normal two chromosome 21s. Trisomy is a specific instance of polysomy, a more general term that indicates having more than two of any given chromosome.
6 TRIPLOIDY
y y y y y y y
triploid individual has three of every chromosome presence of three haploid sets of chromosomes a triploid human would have 69 chromosomes (3 haploid sets of 23) a triploid dog 117 chromosomes production of triploids seems to be relatively common and can occur by, for example, fertilization by two sperm cells. however, birth of a live triploid is extraordinarily rare and such individuals are quite abnormal the rare triploid that survives for more than a few hours after birth is almost certainly a mosaic, having a large proportion of diploid cells.
7 DELETION
y y
chromosome deletion occurs when the chromosome breaks and a piece is lost involves loss of genetic information and results in what could be considered "partial monosomy" for that chromosome
8 INVERSION
y y y
A related abnormality is a chromosome inversion. a break or breaks occur and that fragment of chromosome is inverted and rejoined rather than being lost inversions are thus rearrangements that do not involve loss of genetic material and, unless the breakpoints disrupt an important gene, individuals carrying inversions have a normal phenotype