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Lectures Flashcards

1
Q

What is sex determination?

A

The development of characteristics allowing an individual to be identified as male or female
Reproductive system, phenotype, behaviour, hormones, metabolism

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2
Q

From the bipotential gonad, describe the mechanism of sex differentiation in the male

A

Y chromosome - Testis determining factor encoded by SRY gene - SOX-9 activation - stimulates testis development - steroidogenic factor 1 - Sertoli cells produce AMH to regress female duct - Leydig cells produce testosterone to develop male organs from Wolffian duct

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3
Q

From the bipotential gonad, describe the mechanism of sex differentiation in the female

A

X chromosome - DAX1/Wnt4a activation - stimulates follicular and theca cells in ovary for follicle growth - follicles product oestrogen - oestrogen causes development of female organs from Mullerian duct

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4
Q

In high temperatures what happens to aromatase? What may happen to reptile sexes?

A

Is increased, so increased androgen-oestrogen conversion which increases female offspring or can cause male-female conversion (vice versa in cold)

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5
Q

What is the sex allocation hypothesis?

A

Female mammals are able to adjust offspring sex ratio in response to their maternal condition. I.e. in deer, dominant males father all offspring, subordinate males do not mate. All females get pregnant but only higher rank ones produce sons that can mate. So to maximise reproductive output, dominant females should produce males and lesser females should produce females as all females are impregnated.

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6
Q

What is adaptive control of gender bias?

A

Changing offspring sex bias in response to changing environment - food, population, disease. I.e. if low population density, skews bias towards males as these disperse to fertilise other colonies and maximise genetic potential

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7
Q

Explain the difference between differential fertilisation and differential embyro survival?

A

Differential fertilisation - more male or female sperm selected for fertilisation
Differential embryo survival - more male or female embryos aborted

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8
Q

Name 4 places in the female reproductive tract sperm progression is controlled?

A

Cervix - mucus removes DNA damaged sperm as have poor motility
Uterus - neutrophils remove membrane damaged sperm
Utero-tubal junction - prevents 90% of sperm in uterus entering oviduct to reduce polyspermy
Oviduct - storage in reservoirs and controlled release to give access to oocyte

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9
Q

Explain the control of sperm progression at the UTJ

A

Molecular recognition system in place. Some sperm unable to pass UTJ despite normal motility and morphology due to lack of surface proteins i.e. ADAM1. Lack of these mean unable to bind to ZP.

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10
Q

What is the glycoprotein that causes the negative charge of sperm? What does the negative charge correlate with?

A

CD52 which is involved in sperm binding. Negative charge correlated with DNA integrity and state of maturation so CD52 is a marker of genetic integrity

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11
Q

What is the set of genes controlling acquired immunity? Is this involved in mate choice?

A

Major histocompatibility complex (MHC). Females choose partners with MHC different to their own - more sperm at oocyte when parental MHC differs

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12
Q

How do X and Y sperm differ?

A

By around 3/4% nuclear content, X sperm is longer, different proteins expressed influencing motility and metabolism, individual sperm microRNAs

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13
Q

What is reproductive intervention? Name 5 types.

A

Methods directed at helping individuals to overcome infertility/assist animal production
IVF, AI, ICSI, ET, cloning

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14
Q

What is genetic management?

A

Aiming to avoid inbreeding

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15
Q

What is the extinction vortex?

A

Small population leeds to inbreeding, disease, inbreeding depression, smaller population and extrinction

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16
Q

Explain the difference between in vivo and in vitro embryo transfer

A

In vivo - superovulated cows mate naturally, embryos are flushed from tract and transferred, can be split into multiple
In vitro - ovaries obtained from dead, oocytes extracted and matured then inseminated and transferred when embryo forms

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17
Q

What is cloning?

A

Taking a cell line from a valuable animal and injecting the somatic cell nucleus into an enucleated oocyte from another, which then develops into an embryo in a foster mother

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18
Q

What are the stages or preimplantation development?

A 
2 cell stage
4 cell stage
8 cell stage
Morula
Blastocyst
Hatched blastocyst
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19
Q

When does compaction occur?

A

16-32 cell morula stage

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20
Q

What are the components of the blastocyst?

A

Trophectoderm (outer layer)
Inner cell mass
Blastocoele cavity

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21
Q

What is compaction?

A

The stage preceding blastocyst where cell to cell adhesion increases, outer cells become polarised and form the trophectoderm and inner cells form the inner cell mass, and there is formation of a cavity

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22
Q

What molecule drives compaction?

A

E-cadherin, a transmembrane cell:cell adhesion molecule

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23
Q

Name 4 other molecules important for compaction

A

Calcium
Protein kinase C (relocated E-cadherin)
Alpha catenin
JAM-1 (junctional adhesion molecule)

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24
Q

What are the 2 cell polarisation models at compaction?

A

Inside/outside hypothesis where cell position directs cell fate
Cell polarity hypothesis where cell fate drives the positioning

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25
Q

What factors prevents loss of fluid out of the embryo resulting in expansion to blastocyst?

A

Polarised distribution of Na/K ATPase in the membrane
Established an ion gradient of high Na in blastocoele, facilitating water movement in through aquaporins
Tight junctions preventing loss of fluid

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26
Q

Name 4 key molecules important in the blastocyst, expressed in the ICM or trophectoderm

A

Oct4 - ICM
Nanog - ICM
Gata6 - ICM
Cdx2 - trophectoderm

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27
Q

Explain how energy metabolism differs between oocyte to blastocyst

A

At compaction embryo switches from dependence on TCA cycle in early cleavage, to glycolysis metabolism as blastocyst.
So the ATP energy gained from oxidation decreases and ATP produced increases

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28
Q

State the changes in lactate, glucose and pyruvate uptake/production from oocyte to blastocyst

A

Lactate production increases
Glucose uptake increases
Pyruvate stays around the same - surpasssed by glucose

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29
Q

How does glucose uptake relate to embryo viability?

A

Glucose uptake is higher in a viable embryo.

Very high glycolysis is associated with non viable embryos

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30
Q

What is the optimum O2 level for embryo culture?

Name 3 reasons why

A

5%
20% (atmospheric) leads to more alteration in gene expression, higher amino acid turnover in early cleavage stages and low in blastocyst, lower cell numbers

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31
Q

What functions do amino acids serve? Name 5

A

Essential for embryo development.

Protein synthesis, metabolism, chelation, pH regulation, energy

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32
Q

How does amino acid turnover relate to embryo viability?

A

Embryos with high amino acid turnover at early cleavage stage have low viability compared to those which have steady increase in turnover as they get to blastocyst stage

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33
Q

Name 3 outcomes of abnormal embryo development

A

Fragmentation
Loss of blastomeres
Arrest

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34
Q

In early cleavage stage, faster embryos… (glucose, lactate, glycolysis)

A

High glucose consumption

Low lactate consumption and glycolytic rate

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35
Q

In blastocyst stage, faster embryos… (glucose, lactate, glycolysis)

A

Increased glucose uptake, lactate production and glycolytic rate

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36
Q

Name 3 substrates and 3 products that may be used as biomarkers for embryo viability

A

Glucose, pyruvate, amino acids uptake

Lactate, ammonium, amino acids production

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37
Q

How does oxygen consumption of the embryo change?

A

Increases sharply at blastocyst stage

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38
Q

Define infertility. How many couples are affected?

A

If a woman has not become pregnant after 1 year of regular sexual intercourse without contraception.
Affects 1/6 of couples

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39
Q

What day does fertilisation occur to form zygote?

A

Day 0

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40
Q

What day does first cleavage to 2 cell stage occur?

A

Day 1/2

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41
Q

What day does 4 cell stage occur?

A

Day 2/3

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42
Q

What day does compaction occur? (morula)

A

Day 3/4

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43
Q

What day does blastocyst enter uterus?

A

Day 5

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44
Q

What day does hatching occur?

A

Day 6/7

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45
Q

What are 3 essential factors for embryo implantation?

A

Receptive endometrium
Healthy blastocyst
Communication between mother and baby

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46
Q

Name 3 features of non receptive endometrium

A

Long apical microvilli, high surface negative charge, thick mucin layer

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47
Q

Name 3 features of a receptive endometrium

A

Shorter microvilli, loss of surface negative charge, thinning of mucin coat, formation of pinopodes

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48
Q

What is decidualisation?

A

Postovulatory process of endometrial remodelling in preparation of pregnancy. Includes secretory transformation of uterine glands, influx of NK cells, vascular remodelling. Progesterone driven

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49
Q

What are the 3 stages of embryo implantation?

A

Apposition, adhesion, invasion

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50
Q

What is an inhibitory molecule for attachment?

A

Mucin1

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51
Q

What are promoting molecules for attachment?

A

Leukaemia inhibitory factor, adhesion molecules (E-cadherin, integrins)

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52
Q

What are molecules involved in invasion?

A

MMPs, TIMPs, PGs, COX-2, IL-11, VEGF

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53
Q

What type of implantation occurs in humans?

A

Invasive interstitial implantation

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54
Q

How long is the embryogenic phase?

A

14 days from conceptus to embryo

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55
Q

How long is the embryonic phase?

A

6 weeks from embryo to foetus

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56
Q

How long is the foetal phase?

A

220 days until birth

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57
Q

What does the corpus luteum do?

A

Synthesises progesterone to maintain pregnancy until the placenta takes over at around 12 weeks

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58
Q

What did the dutch famine birth cohort study find?

A

In utero exposure to famine affected the offspring - coronary heart disease in 1st trimester, obstructive airways disease in 2nd and high blood pressure and mental health disease in 3rd

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59
Q

What is the Barker early origins/foetal origins hypothesis?

A

Low weight in infancy increases death from ischaemic heart disease

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60
Q

What does a low birth weight predispose to?

A

CV disease, type 2 diabetes, osteoporosis, depression

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61
Q

What does a high birth weight predispose to?

A

Cancer

62
Q

What is cellular differentiation?

A

Process by which genotype gives rise to phenotype

63
Q

What is epigenetics?

A

Inheritable changes in organisms caused by modification of gene expression rather than changes in the genetic code itself

64
Q

Name 2 types of epigenetic modification

A

Methylation, histone acetylation

65
Q

What is developmental plasticity?

A

Selection of the right phenotype to fit the anticipated future environment

66
Q

What does the maternal-embryonic communication regulate? Name 3

A

Blastocyst morphogenesis
Implantation coordination
Maternal immunotolerance
Developmental plasticity

67
Q

What facilitates sperm transport?

A

Sperm motility and female tract movement

68
Q

What is capacitation?

A

The final maturational stage of sperm that takes place in the female genital tract before sperm gain ability to fertilise oocyte, increasing viability and motility

69
Q

Where is sperm stored in the human and for how long?

A

Sperm can be stored in reservoirs in the fallopian tubes for up to 5 days

70
Q

What is the benefit of sperm storage?

A

Increased motility, viability, delayed time to capacitation so longer lifespan of sperm

71
Q

What are 2 changes that occur in the oviduct in response to sperm presence?

A

Altered protein secretion
Changes in transcription
I.e. adrenomedullin secretion increases at sperm contact with oviduct, increasing ciliary beat frequency

72
Q

What modulates the periconception environment?

A

Short range signals - local responses to gametes

Long range signals - hormones, environment, nutrition

73
Q

What is the purpose of ZP3?

A

Mediates primary sperm binding and acrosome reaction

74
Q

What is the purpose of ZP2?

A

Mediates secondary sperm binding

75
Q

What is the purpose of ZP1?

A

Forms a crosslink between 2 and 3 as a protein network

76
Q

What is the cortical reaction?

A

Occurs when a sperm binds to the oocyte membrane. Increased calcium results in mobilisation of cortical granules to the membrane, and release into perivitelline space, where they act on ZP to prevent further binding

77
Q

What is the acrosome reaction?

A

Occurs when a sperm binds to the ZP. Acrosome swells, membrane binds to the overlying plasma membrane, large increase in Ca2+ and contents of acrosomal vesicle are exocytosed out. These enzymes digest the ZP to allow sperm to reach oocyte

78
Q

What are 3 patterns of fertility behaviour?

A

Starting, stopping, spacing

79
Q

What is fecundity?

A

Physiological capability of a woman to produce a live birth

80
Q

What is fertility?

A

How many live births actually achieved by a woman

81
Q

What is prospective cohort analysis?

A

How many children a woman has over her lifetime, following her as it happens i.e. by population register

82
Q

What is retrospective cohort analysis?

A

How many children a woman has HAD over her lifetime, collected i.e. by medical history or survey after childbearing age

83
Q

What is period analysis?

A

Number of births in a particular time period, collected i.e. by birth register

84
Q

What is parity progression ratio?

A

Cohort measure

Proportion of women with at least ‘n’ children who go on to have at least one more child ‘n+1’ (expressed P01, P12)

85
Q

What is crude birth rate?

A

Period measure
Ratio of live births in a time period i.e. one year to the average population in that period, expressed as births per 1000 population
(births/population x1000)

86
Q

What is general fertility rate?

A

Period measure

Births per 1000 women aged 15-49

87
Q

What is age specific fertility rate?

A

Period measure

Births per 1000 women aged a certain age (x)

88
Q

What is total fertility rate?

A

Period measure expressed as cohort measure - synthetic cohort measure
Number of children who would be born to a woman if she lived to the end of her childbearing years and had children in accordance with current age specific fertility rates

89
Q

What are 5 lowest fertility countries?

A

Taiwan, Macao, Hong Kong, Singapore, South Korea, Moldova, Bosnia and Herzegovina, Portugal, Spain, Hungary

90
Q

What are 5 highest fertility countries?

A

Niger, Somali, Mali, Chad, Angola, Congo, Burundi, Uganda, Timor-Leste, Gambia

91
Q

What is immunology?

A

Study of the defence mechanisms of an organism

92
Q

Name the types of pathogen

A

Bacteria, virus, fungi, parasites

93
Q

What is immunity?

A

Protection from infectious disease

94
Q

What are the 3 branches of the immune system?

A

Physical and chemical barriers
Innate immune system
Adaptive immune system

95
Q

What are the 5 signs of inflammation?

A

Heat, redness, swelling, pain, loss of function

96
Q

What are 4 aspects of the innate immune system?

A

Physical barriers - skin
Immediate action - anti microbial peptides
Non specific - pattern recognition receptors
Leukocytes - neutrophils, NK cells, dendritic cells, macrophages

97
Q

What are pattern recognition receptors?

A

Germ line encoded receptors which can recognise pathogen associated molecular patterns (PAMPs) synthesised by microorganisms

98
Q

What are toll like receptors?

A

PRRs which recognise PAMPs and initiate downstream signalling events, leading to release of cytokines/chemokines/anti microbial peptides which cause recruitment of neutrophils and macrophages and maturation of dendritic cells

99
Q

What are 2 aspects of the adaptive immune system?

A

Cellular effectors - cytotoxic/helper T lymphocytes

Humoral effectors - B lymphocytes secrete antibodies

100
Q

Name 3 differences between the innate and adaptive immune system

A

Innate - early recognition, low specificity, no memory

Adaptive - late recognition, specific, long memory

101
Q

What are potential sources for infection of the female tract?

A

Menstruation, fertilisation, implantation, pregnancy, parturition

102
Q

How does the innate immune system facilitate events of the female tract?

A

Helps passage of sperm
Facilitates implantation
Enables pregnancy to continue

103
Q

How does oestradiol affect immunity?

A

Increases susceptibility to infection, allows sperm into the tract

104
Q

How does progesterone affect immunity?

A

Increases immune surveillance

105
Q

What are the 3 immunological phases of pregnancy?

A

1 - implantation and placentation resemble an open wound and elicit strong inflammatory response
2 - growth in the second trimester
3 - renewed inflammation promoting contractions and birth

106
Q

How does immune system respond to implantation in the decidua?

A

DCs, NK cells, macrophages infiltrate decidua and accumulate around embryo, without these there is negative effect on development

107
Q

What is immune tolerance?

A

Reducing impact of a pathogen without actually fighting it - i.e. pregnancy

108
Q

Describe the grading system of early cleavage embryos

A

3 numbers representing the number of cells, degree of ideal cells, and degree of fragmentation (higher numbers better)
8, 4, 4 = best quality 8 cell embryo
5, 1, 1 = worst quality 5 cell embryo

109
Q

Describe the grading system of blastocyst embryos

A

Grade 1-4 for degree of expansion and 5-6 for hatching (higher numbers better)
If 3 or higher, further graded A-D for ICM and trophectoderm (A highest)

110
Q

What is implantation?

A

Series of stages including blastocyst hatching, adhesion, endometrial invasion, embryo differentiation and growth to produce a state of clinical pregnancy

111
Q

What is implantation rate?

A

Number of embryos implanted over number transferred in a given time period

112
Q

What is repeated implantation failure?

A

Failure to reach clinical pregnancy (gestational sac/foetal heartbeat on USS) after the transfer of at least 4, good quality fresh or frozen embryos in a women <40 over at least 3 transfer cycles

113
Q

When is the implantation window?

A

Day 6-10 after fertilisation

114
Q

What cell types does the trophoblast differentiate into?

A

Cytotrophoblast

Syncytiotrophoblast

115
Q

What is the objective of ART?

A

To bring sperm and oocyte closer together to increase chances of fertilisation and pregnancy

116
Q

Give 3 reasons for ART

A

Infertility, absence of a partner, genetic engineering

117
Q

State each step of the IVF-ET procedure

A

Superovulation and egg collection - semen collection - insemination - fertilisation check - embryo culture - embryo transfer (/cryopreservation) - luteal support - pregnancy test

118
Q

State the procedure of oocyte collection

A

Monitor follicular development by TVUSS, and when at least 2 follicles are 17mm diameter give hCG to induce maturation and aspirate with a catheter

119
Q

Name methods of sperm selection

A

Percoll swim up
DNA probe stains - SYBR14
Flow cytometry
Fluorescence activated cell sorting

120
Q

When is ICSI used?

A

If sperm are unable to fertilise - abnormal morphology, motility, damaged acrosome, immature

121
Q

What is PGD?

A

Pre implantation genetic diagnosis. Used to screen embryos for genetic disorders

122
Q

How is PGD performed?

A

At 8 cell stage (2/3 days), zona drilling performed to remove 1/2 blastomeres to screen for disease

123
Q

What is CRISPR-Cas9 used for?

A

Enables gene editing for identification, removal and replacement of faulty genes

124
Q

Explain how CRISPR-Cas9 works

A

Clustered regularly interspaced short palindromic repeats - CRISPR - RNA sequence generated to reflect sequence of target gene
CRISPR RNA coupled with the restriction enzyme Cas9
CRISPR-Cas9 complex injected into cell
Target sequence snipped out of DNA by Cas9 to disable gene
Synthetic DNA can be spliced at the restriction zone to introduce a desired trait

125
Q

How much semen and sperm do men ejaculate?

A

Men ejaculate 5ml of semen with 250 million sperm

126
Q

What is migration efficiency?

A

The distance (mm) travelled by sperm in 7 minutes

127
Q

What 2 things does high fertility need?

A

Competent sperm

Sufficient duration

128
Q

What are ROS?

A

Reactive oxygen species, important in capacitation/acrosome reaction/binding but oxidative stress can cause damage to sperm DNA

129
Q

What is cryopreservation?

A

When a substance is preserved by cooling to very low temperatures to stop any processes that would cause functional or material damage

130
Q

What are the 3 steps in cryopreservation?

A

Cooling
Freezing
Thawing

131
Q

What are the 4 requirements of sperm cryopreservation?

A

Retain an intact plasma membrane
Cell function should not be impaired
Organelles should be intact and functional
DNA should be intact

132
Q

How does cryopreservation affect sperm?

A

Reduces number of surviving sperm and their competency - damages membranes
Shortened lifespan - premature capacitation - so insemination must be accurate

133
Q

What is vitrification?

A

Rapid cooling to avoid ice crystals, typically involving very high levels of cryoprotectants so sperm do not survive

134
Q

What is a stem cell?

A

Relatively primitive cell that is capable of self renewal, potency and differentiation

135
Q

Give 2 functions of stem cells in the body

A

Development and regeneration

136
Q

Name 4 types of embryonic stem cell

A

Extraembryonic endoderm stem cells (XEN)
Trophoblast stem cells (TS)
Human naive embryonic stem cells (hESCs)
Epiblast stem cells (EpiSCs)

137
Q

Name 3 types of stem cell other than embryonic

A

Foetal stem cells
Adult tissue specific stem cells
Induced pluripotent stem cells

138
Q

Give 4 applications of stem cells

A

Regenerative medicine, drug discovery, toxicology, disease modelling

139
Q

How are human embryonic stem cells derived?

A

Blastocyst grown in IVF medium, immunosurgery with anti-trophectoderm antibodies and complement, inner cell mass isolated and cultured

140
Q

What are 3 fates of stem cells?

A

Self renewal, differentiation, apoptosis

141
Q

What can induced pluripotent cells be used for?

A

Can capture a specific genome from a population/disease and study effects
Can test drugs on normal and affected phenotypes to see if they would be effective in specific populations

142
Q

What are the 3 factors needed for successful stem cell treatment of disease?

A

Make desired cell, transplant it correctly and make sure it integrates

143
Q

Name 5 ARTs for male factor infertility

A

IUI, ICSI, IVF, SSR, DI

144
Q

What are the 2 types of pituitary suppression? What are the pros/cons?

A

GnRH agonists - may cause OHSS, longer and more expensive

GnRH antagonists - no initial flare up, shorter treatments, more flexibility and reversibility

145
Q

How are follicles stimulated?

A

FSH daily injection given to stimulate oocyte growth

146
Q

How is ovulation triggered? Is there a risk?

A

hCG injection when follicles are big enough - may have a risk of OHSS (or GnRH agonist if using an antagonist regime, less OHSS risk)

147
Q

What is the preferred number of embryos transferred? When is this changed?

A

Single embryo transfer
2 transferred - if over 40 straightaway, if 37-39 only if there is not top quality embryos in 1st/2nd cycle (or always if 3rd cycle), and if under 37 transfer 2 on 3rd cycle and if on 2nd cycle there is not top quality embryos

148
Q

What is luteal support?

A

Progesterone (or hCG) injections in the first few weeks to maintain pregnancy

149
Q

Give 5 factors affecting IVF success

A 
Age
Cause of infertility
Duration of infertility
Number of previous attempts
Other medical conditions
Environmental factors
Obesity
150
Q

Explain the difference between action of GnRH agonists and antagonists and when they are given

A

GnRH agonists given continuously, suppressing gonadotrophins after an initial surge, then FSH stimulation performed and maturation triggered
GnRH antagonists suppress gonadotrophins immediately, given after FSH stimulation and before maturation triggered

Embryo (1 decks)

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