Lesson 1 Introduction.
The Chromosome Theory of Inheritance
-In the 1800s German scientist Walther Flemming made fire observations of darkly stained
threadlike structures in the nucleus of the cell. (eventually named chromosomes). Also the first
to describe cell division.
-Theordor Boveri and Walter Sutton set up the basis for chromosome theory of inheritance
and the field of cytogenetics.
-Sutton stupied great lubber grasshopper(Brachystola magna) he concluded that X and Y
chromosomes determine the sex of species this study was incomplete therefore non conclusive.
-Thomas Hunt Morgan’s fruit fly lab study proved this (kinda).
Fast Forward
-Fruit flies eye colour is determined by a gene called “white”, that produces an enzyme that
makes red pigment(eyes). Capital “W” means the gene is present(red eyes).
-Alleles: Different versions of the game genes.
Chromosome Terminology
-At the mitotic metaphase chromosomes are at their most condensed state
-diploid cells have two copies of each chromosome called homologous chromosomes.
-Each chromosome consists of two identical chromatids called sister chromatids because each
chromosome was duplicated during the S phase of the cell cycle.
-Every chromosome has a centromere located in the middle.
Chrom_____?
Chromosome: Threadlike structure of nucleic acids and protein found in the nucleus of most
living cells carrying genetic information in the form of genes.
Chromatid: Each of the two threadlike strands into which a chromosome divides longitudinally
during cell division.Each contains a double helix of DNA.
Chromatin:The material of which the chromosomes of organisms other than bacteria are
composed. It consists of protein,RNA,and DNA.
-Each chromosome in the pair will have the same genes
-homologous chromosomes:Each chromosome in the pair will have the same genes but not
necessarily the same version.
Nonhomologous chromosomes:Chromosomes that do not belong to the same pair.
Haploid vs Diploid
-Meiosis only occurs in organisms that reproduce sexually.
-gametes(sperm cells) are produced by parents and contain half the number of chromosomes in
their genome.
-zygote is when the gametes fuse together (sex cells).
-individuals with a single copy of each different chromosomes are called haploid
-diploid is when the sperm and ova fuse during fertilization resulting in two complete sets of
chromosomes.
Heterozygous:Having two different alleles of a particular gene (Ex. Aa)
Genotype:The complete set of an organism's genetic material. The complete set of alleles
Phenotype:Is what we see. The visible or detectable effect of the alleles.
Karyotypes
Autosomes: Any chromosomes that are not sex chromsomes (humans have 22 pairs of
autosomes.
Sex Chromosomes: (X and Y).
Karyotype: A process by which photographs of chromosomes are taken in order to determine
chromosome complement of an individual
-To create karyotypes, chromosomes from cells in metaphase are isolated and spread out in a
microscope slide, then they are stained photographs and then a computer program arranges the
chromosomes into pairs.
-Karyotypes can be used to identify a species based on the number and staining pattern of the
chromosomes,or to determine an organism ploidy. They can also be used to determine if any
gross chromosomal differences(male vs. female).
-Cri-du-Chat syndrome part of chromosome #5 is missing(deletion)
-In Down syndrome, three chromosome #21’s will be present(trisomy-21)
-Four different types of chromosomes
-Metacentric(Centromere in middle)
-Submetacentric(Centromeme off centre
-Acrocentric(centromere near one end)
-Telecontric(Centromere is at one end)
Lesson 2:The Cell Cycle and C-Value
The Cell Cycle Overview
-The cell cycle is a way to describe the process of cell growth and division
-Stages G1,G0,S and G2 are collectively called interphase.
-M phrase represents mitosis(division of chromosomes) and cytokinesis(cell division).
-Most of the growth of a cell occurs during G1 and G2.
-G is the gap phase sometimes referred to as the “growth” phase.
-Certain cells in our bodies are nondividing, like our nerve cells.
-These cells are said to be in G0 phase.
-The S phase is named because this is when DNA synthesis takes place
-DNA replicates and doubles in amount to prepare for mitosis
S has 2 sets
G2 has 2 sets
The Cell Cycle S Phrase
-During S phase the DNA is replicated
-In G1 Chromosomes contain a single double-stranded DNA(dsDNA) molecule
-In S phase each (dsDNA) is replicated to produce two identical copies; these copies are held
together to create chromosomes that now have two dsDNA molecules per chromosome.These
two dsDNA molecules are now contained within sister chromatids.even though the amount of
DNA in the cell doubled the number of chromosomes remains the same.
-In G1 phase each chromosome contains a single double stranded DNA molecule following
DNA replication in S phase each chromosome contains two double stranded DNA
molecules(sister chromatids).
-Sister chromatids are held together at the centromere by a protein called cohesin; they are still
one structure which is why they are still referred to aws one chromosome.
C-Value:
-In a haploid cell (G1 phase) there is only one chromatid per chromosome so there is only one
copy of the genome (1c) in a haploid cell in G1 phase.
-Following S phase, the DNA has replicated and now each chromosome has two sister
chromatids per chromosome Now there are two copies of the genome so the cell is 2c
C-Value Paradox And Gene Density
-C-value is a measure of the amount of DNA present in the nucleus of a haploid cell (1C)
usually measured in picograms.
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Lesson 3:Mitosis
Overview of Mitosis
-Mitosis is a nuclear division that gives rise to two daughter cells that have the same number of
chromosomes as the parent cell. 5 stages of mitosis.
Prophase
Chromosomes begin to condense and become visible.
-Normally it is impossible to discern chromosomes in the nucleus of a cell.
-As the chromosomes condense the nucleoli disappear and the centrosomes (made up of two
barrel-like centrioles begin moving to opposite poles.
-spindle microtubules emerge from the centrosomes.This now forms the mitotic spindle
apparatus, which is composed of three kinds of microtubules: astral, kinetochore and interpolar.
Prometaphase:
-Kinetochores begin to form on the centromeres of each sister chromatid soon after S-phase.
-When the cells reach prometaphase the kinetochore contains more than 50 different proteins.
-At the start of prometaphase the nuclear envelope breaks down and microtubules from the
centrosomes move and connect with the kinetochores at the centromeres of each sister
chromatid the forms the mitotic spindle apparatus which is composed of three kinds of
microtubules:astral,kinetochore and interpolar.
-The kinetochore microtubules' role is to position the chromosomes along an imaginary plate in
the middle of a cell called the metaphase plate.
-The astral microtubules help stabilize the spindle apparatus by anchoring it to the cell
membrane.
-The interpolar microtubules project from each spindle pole crossing the equation of the cell
from opposite sides here motor proteins attach and the interpolar microtubules slide along one
another to push the spindle poles to opposite sides of the cell and then later to elongate the cell
to prepare for cytokinesis(cell division).
-The sister chromatids are actually glued together along their entire length by a protein called
cohesin( A chromosome-associated multisubunit protein complex that is highly conserved in
eukaryotes)
-Just before prometaphase an enzyme called separase is released which breaks down the
cohesin proteins which allows the sister chromatids to separate but not completely.
-At the centromere regions cohesin is protected by another protein called shugoshin which
keeps the sister chromatids together during prometaphase and metaphase.
Metaphase:
-Microtubules move chromosomes to the metaphase plate with each sister chromatid attached
to microtubules from opposite poles.
-The sister chromatids are still held together in the centromere region by cohesin with the help
of shugoshin
-The microtubules set up an equilibrium of pushing the chromosomes from opposite poles which
results in them lining up at the metaphase plate.
-once equilibrium is achieved with each chromosome sitting on the metaphase plate,
metaphase is complete.
Anaphase:
-Anaphase happens so fast
-Once the chromosomes are fully aligned at the end of metaphase,shugoshin comes off the
centromere region and the remaining cohesin is now degraded by an enzyme called separase.
-Once the chromosomes are fully aligned at the end of metaphase, shugoshin comes off the
centromere region and the remaining cohesin is now degraded by an enzyme called separase.
-Sister chromatids are then pulled to opposite poles by the kinetochore microtubules. As the
chromatids are pulled across the cytoplasm they form a v shape
Telophase:
-after anaphase the chromosomes are at opposite poles and begin to deconsendes
-Nuclear envelopes begging to form around these two clusters of chromosomes
-Soon followed by the appearance of nucleoli
-The spindle fibres break down and the chromosomes decondense.
Cytokinesis
-Final step where the cell physically divides into two
-In animal cells a contractile ring of actin microfilaments forms in the middle of the parent cell
and slowly contracts to form a cleavage furrow eventually pinching off forming two daughter
cells.
-In contrast,plant cells have rigid cell walls and build a new cell wall between the two nuclei
called a cell plate.
The organelles are also distributed to each daughter cell.
Meiosis Introduction
-The main purpose of meiosis in a diploid organism is to create gametes.
-gametes fuse to form diploid zygotes
-in the first division (Meiosis i) homologous chromosomes are separated from one another and
in the second(meiosis ii) division sister chromatids are separated from one another essentially
like it occurs in mitosis.
-meiosis I introduces two sources of genetic variation, during pairing of the homologues during
prophase I, recombination will lead to new alleic combinations on the chromosomes, then in
metaphase I the arrangement of the homologous on metaphase I plate will ead to random
assorting of the chromosomes to the daughter cells
Meiosis
-Stars with a diploid cell that is in G2 cell cycle which means each chromosome is made up of 2
sister chromatids.
-In a diploid cell in G2 phase thereof 4C (copies of the genome) which are used to produce 4
haploids gametes each with 1C one copy.
Meiosis I Phases:Prophase I
-Prophase I is subdivided into 5 substages: Leptotene,Zygotene,Pachytene,Diplotene and
Diakinesis.
Leptotene:Chromosomes begin to become more condensed, the centrosomes move to
opposite poles to begin setting up the spindle apparatus
Zygotene:The homologous chromosomes begin to pair up. A process involving a series of
DNA-DNA comparisons that pairs up the homologs in a process called synapsis. Each paired
chromosome is held together by a protein complex called the synaptonemal complex. When the
two homologous pairs are paired together there are now four chromatids in this complex called
a tetrad.
Pachytene:In this stage recombination occurs, this process rearranges genetic information from
the maternal and paternal copies of each pair of synapse homologous. Following pachytene
each homolog will look like a “harlequin” with patches of maternal and paternal segments along
each homologue. This contributes to genetic variation in the offspring.
Diplotene
-During Diplotene the synaptonemal complex breaks down, when this happens the crossover
points,called chiasmata can be seen between the chromosomes.now that the synaptonemal
complex is gone the chiasmata holds the non-sister chromatids together.
Diakinesis
-The nuclear membrane breaks down, the spindle apparatus continues to form and the
chiasmata terminalize (move to the telomere regions of the chromosomes). During both
diplotene and diakinesis the chromosomes become more and more condensed to prepare for
their movement to the metaphase I plate.
Metaphase I and Anaphase I
-Spindle microtubules from one pole attach to one chromosome of a homologous pair and
spindle microtubules from the other pole attach to the other homologous chromosome of the
homologous pair(tetrad) and each pair aligns along the metaphase I plate. This set is a major
source of genetic variation due to what Mendel termed to be an independent
assortment.Independent assortment occurs since each pair of homologs can sit on the
Metaphase I plate in one of two orientations (see Figure 4.4). If someone has a genotype of
AaBb with the “A” and “B” genes on different chromosomes, then the “A” and “B” genes will
assort independently of one another. This means that whether a gamete will receive the A or an
allele is not dependent on which other alleles they receive.For each gene that is heterozygous,
there are two different types of possible gametes. So, with the AaBb individual, there will be 2 x
2 = 4 different gametes produced due to independent assortment. The genotypes of the
possible gametes are AB, ab, Ab, and aB which will be observed in a 1:1:1:1 ratio (Figure 4.4).
Humans have 23 pairs of chromosomes which results in over 8 million different gamete
combinations! A general rule for diploid organisms is that the number of different gametes is
equal to 2n, where n equals the number of different chromosomes.
Anaphase I: During anaphase I the two chromosomes that make up the homologous pairs are
separated and pulled to opposite poles
-This results in a haploid set of chromosomes being delivered to each pole of the spindle in
other words 2n to n
-Meiosis I is sometimes called the reductional division due to this halving in chromosome
number.
Telophase I
Once the chromosomes have finished migrating to the poles, telophase I begins. The spindle
disassembles, the nucleus reforms and cytokinesis divides each cell into two haploid daughter
cells. In humans, these haploid daughter cells will be n=23. It is important to note that at the end
of Meiosis I each chromosome is still in the replicated state, with two sister chromatids per
chromosome. Unlike in mitosis, the two daughter cells that are produced at the end of Meiosis I
are not genetically identical due to crossing over and independent assortment.
Meiosis II
-All the stages of meiosis II are the same as mitosis.
-Here the chromosomes align on the metaphase II plate individuals and during anaphase II, the
sister chromatids are pulled to opposite poles by the spindle microtubules
-Following telophase II and cytokinesis the four haploid cells produced will all be genetically
distinct.
Comparing Mitosis and Meiosis
-Mitosis results in two genetically identical daughter cells that have the same number of
chromosomes as the parent cell.
-THe goal of mitosis is to produce more cells for growth in contrast meiosis begins with a diploid
parent cell and produces four haploid daughter cells which are all genetically unique (differences
arise in meiosis I).
-In prophase I homologous chromosomes pair with each other allowing for genetic
recombination and in metaphase I, each pair of homologues can align on the metaphase I plate
in two different orientations giving rise to independent assortment
-in meiosis I the chromosome number is halved during anaphase I when the homologous are
pulled to opposite poles.
-The products of meiosis are only ever used for sexual reproduction.
Recombination
In Dna recombination two DNA strands that are crossing over are cut and then joined,
-in certain fungi (yeast and bread mold), the products of meiosis (haploid spores) are held
together in a sac (ascus). Yeast can also grow as a haploid, so we can determine the
phenotype of these spores, which we can’t do easily in animal species like ourselves. Let’s say
we look at a gene that is involved in spore colour, that we’ll call the “D gene”. The “D” allele
results in darkly coloured spores, while the “d” allele results in lightly coloured spores.
-If you look at the meiotic products of a diploid yeast cell that is Dd, you would expect to see in
each ascus ½ dark and ½ light spores (we call this a 2:2 segregation pattern). However, in
some of the asci you will find ¾ dark, ¼ light and in others, ¼ dark, ¾ light spores (these are 3:1
or 1:3 segregation patterns) (Figure 4.8).
-What happened to the other allele in the 3:1 and 1:3 asci? It looks like in the 3:1 pattern that
one of the “d” alleles was converted into a “D” allele, and that one of the “D” alleles was
converted into a “d” allele in the 1:3 asci. This is what is referred to as gene conversion and it
provides some insight into how recombination occurs.
-In the 1960s Robin Holliday came up with a model for recombination
-When a crossover occurs a single strand cut is made in the DNA on two non sister chromatids
of a homologous pair which then reattach to one another to create a crossover.
-The exchange point migrates down the chromosomes pulling the top strand down and the
bottom strand up this creates a patch of heteroduplex DNA on both non-sister chromatids
-Any differences will be repaired soon after but it is generally random which base will be
replaced.
Lession 5:Sex Determination in Humans
Turner and Klinefelter Syndromes
-Henry Turned described a small number of human females that had abnormal sexual
development like under 5 feet,skin flaps on the back of the neck, a shield-like chest and
underdeveloped secondary female sexual characteristics.
-Many of these individuals were sterile this was termed Turned syndrome individuals had a
single x chromosome 2n=45 XO
-Harry Klinefelther described a similar thing in males; he determined it to be Klinefelter
syndrome 2n=47 and phenotypically male. They have extra x chromosome,
-generally taller than average with long legs and arms their tests are small than normal which is
contributing factor to lack of sperm production in half of klinefelter males, for males with sperm
there sterility is usually low in addition some feminine sexual development occurs resulting in
slight breast development and rounding of hips
-Klinefelter affects 1/660 males and in turn is present in 1in every 2000 females. Both errors
occur due to the same errors in the separation of chromosomes during meiosis in a process
called nondisjunction the reason for the differences is because a lot of the 2n=45 XO die in
utero
-Nondisjunction is the failure to separate either the homologous in meiosis I or the sister
chromatids in meiosis II leading to the gametes having an extra chromosome missing.
XXX Syndrome and XYY Condition
-Females with 3 X (XXX) are not identified unless they have a karyotype test
-Some Triplo-X individuals have underdeveloped secondary sex charactics,sterility and
deplayed language and motor functions.
XYY Condition
-Nondisjunction events in males with the Y chromosome in meiosis II lead to male offspring that
are 2n=47
-they are significantly above average in height and certain studies have shown behavioural
problems.
Sex Determination In Humans
-One regions called SRY(Sex-Determining Region Y) is all that is needed to determiner whether
the gondadal tissue will develop into testes.
-A gene in this region codes for a protein called testis-determining factor(TDF) that stimulates
the gonadal tissue to form tests which then produce testosterone which triggers a series of
changes in gene expression to result in a male.
-Swyer syndrome are people with a female phenotypically yet their karyotypes show they are
genotypically XY
-20% of those swyer syndrome the SRY region is either absent or nonfunctional on the Y
chromosome
-The other 80% have disruptions in one or more of the key transition points along the pathway
to producing testes.
-those with swyer syndrome don’t develop female secondary characteristics and must undergo
horm-The SRY regions still causes thone therapy.
- Another Phenotypically genotypically XY is called Androgen Insensitivity Syndrome females
with AIS have a functional SRY regions but they lack the cell receptor for testosterone and other
androgens
e formation of testes but they are usually reduced in size and location
-The tests still product testosterone but without the cell receptor changes in gene expression
that are normally triggered by testosterone resulting in male development dont occur
X Chromosome Inactivation
-Klinefelter and Turner syndromes are examples of aneuploidy
-The only individuals with an extra autosome are those with down syndrome (2n 47,21,21,21 or
Trisomy-21)
The Lyon Hypothesis
-Mary Lyon and Liane Russel suggested that Barr bodies represented an X-chromosome that
was “shut down”.
-The general rule is that number of Barr bodies is equal to the number of X-chromosomes minus
1
-The X-chromosome that is inactivated is randomly selected in a given cell but once it is
shutdown all progenitor cells will have the same X-chromosome inactivated.
-If the female is heterozygous for genes on the X-chromosomes,then she will be a mosaic with
half of her cells expressing the gene and the other half not.
-The Calico cat is also genotypically XBXO with black and orange patches. In addition, it has
white patches which are due to another gene located on one of the autosomes that controls
melanocyte migration (the Spotting allele).