From Embryology

--Mark Hill (talk) 17:16, 5 August 2015 (AEST) Well done, you were first! We will be talking more about this in the Practical on Friday.

Please do not use your real name on this website, use only your student number.

2015 Course: Week 2 Lecture 1 Lecture 2 Lab 1 | Week 3 Lecture 3 Lecture 4 Lab 2 | Week 4 Lecture 5 Lecture 6 Lab 3 | Week 5 Lecture 7 Lecture 8 Lab 4 | Week 6 Lecture 9 Lecture 10 Lab 5 | Week 7 Lecture 11 Lecture 12 Lab 6 | Week 8 Lecture 13 Lecture 14 Lab 7 | Week 9 Lecture 15 Lecture 16 Lab 8 | Week 10 Lecture 17 Lecture 18 Lab 9 | Week 11 Lecture 19 Lecture 20 Lab 10 | Week 12 Lecture 21 Lecture 22 Lab 11 | Week 13 Lecture 23 Lecture 24 Lab 12 | 2015 Projects: Three Person Embryos | Ovarian Hyper-stimulation Syndrome | Polycystic Ovarian Syndrome | Male Infertility | Oncofertility | Preimplantation Genetic Diagnosis | Students | Student Designed Quiz Questions | Moodle page

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Test student 2015

Lab Attendance

Lab 1 --Z5088434 (talk) 13:46, 7 August 2015 (AEST)

Lab 2 --Z5088434 (talk) 12:07, 14 August 2015 (AEST)

Lab 3 --Z5088434 (talk) 12:21, 21 August 2015 (AEST)

Lab 4 --Z5088434 (talk) 12:12, 28 August 2015 (AEST)

Lab 5 --Z5088434 (talk) 12:05, 4 September 2015 (AEST)

Lab 6 --Z5088434 (talk) 12:12, 11 September 2015 (AEST)

Lab 7 --Z5088434 (talk) 12:16, 18 September 2015 (AEST)

Lab 8 --Z5088434 (talk) 12:37, 25 September 2015 (AEST)

Lab 9 --Z5088434 (talk) 13:55, 9 October 2015 (AEDT)

Lab 10 --Z5088434 (talk) 12:07, 16 October 2015 (AEDT)

Lab 11 --Z5088434 (talk) 13:17, 23 October 2015 (AEDT)

Lab 12 --Z5088434 (talk) 12:02, 30 October 2015 (AEDT)

Lab 1 - Online Assessment

Summary of Article I

Low-dose growth hormone supplementation increases clinical pregnancy rate in poor responders undergoing in vitro fertilisation[1]

PMID 26193891

The aim of the prospective, self-controlled study was to investigate the impact of low-dose growth hormone (GH) supplementation for patients undergoing in vitro fertilization (IVF) with poor ovarian response (POR), which is defined as IVF incidence rates between 9 and 24%. GH is known to participate in follicular development by regulating gonadotropins in granulosa cells which in turn regulate the synthesis of IGF-I. This growth factor plays a major role in the synthesis of sex steroids and, thus, oocyte maturation. GH has been administered to POR patients since the 1990s, however, with mixed, mostly positive, results. Therapeutic advantages were largely established in women of advanced reproductive age. A possible reason for this unsuccessful administration is proposed to be the GH dose; the majority of doses used for POR patients is equal to those used in patients with GH-deficiency. However, POR patients are usually not GH-deficient, which leads to the investigation of the effects of lower dose GH supplementation to POR patients. A lower dosage is safer, since occurrence of side effects correlates with the dose of GH. In addition, it offers a more economic treatment strategy.

This study recruited 64 women with history of POR and an absence of pregnancy in at least two previous IVF cycles who were given GH during their third cycle. Certain factors, such as high BMI, different diseases, or surgeries lead to exclusion of the prospectus patient. The included patients underwent an ovarian stimulation protocol. This protocol involved ovarian hyperstimulation using a GnRH agonist to control for any differences besides Non-GH- and GH-cycle. A dose of 0.5 IU GH was supplemented while the GnRH agonist was given until human chorionic gonadotropin (hCG) was administered. 36h after hCG administration oocytes were retrieved and standard IVF procedures were performed. The clinical pregnancy rates, the number of retrieved oocytes, and obtained embryos, embryo quality, and cycle cancellation rate of the Non-GH- and the GH-cycle were then statistically analyzed.

Even though a greater number of oocytes and embryos were obtained in the GH-cycles and the cycle cancellation rate was lower in GH-cycles, the differences were not statistically significant. However, the number of top quality embryos obtained was significantly higher in the GH-cycles. The clinical pregnancy rate for the GH-cycles was 34.4%. These results align with prior knowledge about the mechanism and role of GH for follicular development. The evidence for ovarian GH receptors and this study's observation of its correlation with improved pregnancy rates and embryo quality may indicate that GH effects oocyte maturation. Additionally, low GH dosage was proven to be successful for POR patients' IVF outcome and, thus, may offer a safer and more economic treatment strategy.

Summary of Article II

Artificial oocyte activation in intracytoplasmic sperm injection cycles using testicular sperm in human in vitro fertilization[2]

PMID 26161332

The study aims to evaluate artificial oocyte activation (AOA) with a calcium ionophere as an effective method for severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. In the case of male factor infertility intracytoplasmic sperm injection (ICSI) is applied. The majority of failures of this method can be traced back to oocytes remaining inactivated despite appropriate injection of spermatozoa, with more than 80% of unfertilized oocytes being arrested at metaphase II stage. During oocyte activation intracellular calcium concentrations rise drastically in form of calcium oscillations. These oscillations are presumed to be triggered by certain spermatozoa factors delivered to the oocyte upon membrane fusion and cause the resumption of meiosis and multiple events of oocyte activation. In cases of non-motile spermatozoa PF is commonly used to induce motility and AOA is applied in cases where PF does not restore motility. However, the combination of both fertility methods is relatively unexplored. This study, therefore, explores the combined efficiency of PF and AOA on fertilization and pregnancy rates after ICSI.

29 patients were included who underwent AOA with a calcium ionophore after ICSI. In addition, a control group of 480 patients who only underwent ICSI without AOA was included in the study. All ICSI cycles involved male factor infertility. Oocytes and testicular spermatozoa extraction were conducted using conventional IVF methods. Non-motile spermatozoa were treated with 5mM PF to induce motility and were injected into the oocytes. 30 minutes post-ICSI the oocytes were exposed to 10uM calcium ionophere for 5 minutes and conventional IVF procedures were resumed. Embryo quality, pregnancy, and delivery rate were statistically analyzed. In addition, the effects of AOA and PF were assessed individually.

The quality of embryos was significantly lower in the AOA group compared with the control group. Similarly, delivery rates were lower in the AOA group than in the control group. Fertilization rates, however, did not account for significant differences. Prior studies have shown that DNA damage and constrained spermatozoa motility are negatively related; this potentially accounts for the decreased embryo quality in the AOA group. Within the AOA group in 17 cases sperm motility was not restored after PF exposure. Nevertheless, there was no difference between the motile and non-motile spermatozoa in fertilization, pregnancy, or delivery rate. AOA may, therefore, be useful in patients with low fertilization rate or total failure fertilization rate as it ensures fertilization regardless of the success of PF treatment.

--Mark Hill (talk) 17:38, 3 September 2015 (AEST) These are good summaries of the papers. (5/5)

Lab 2 - Online Assessment

Different Stages of Embryo Development.jpeg

Different Stages of Embryo Development[3]

PMID 25935518

--Mark Hill (talk) 17:40, 3 September 2015 (AEST) Image uploaded with reference, copyright and student template. Would have been good if you had also indicated that this was a human embryo somewhere. (5/5)

Lab 3 - Online Assessment

<pubmed>26168107</pubmed> This article reviews the cytogenetic techniques and embryo biopsies required for PGD & PGS and gives an account on the differences in PGD for single gene defects and chromosomal translocations.

<pubmed>22723007</pubmed> This article gives relatively recent and detailed information on the three types of biopsy performed on embryos at different stages of development (before conception, after fertilization, and early cleavage or blastocyst stage)

<pubmed>24515905</pubmed> This article reviews indications for PGD focusing on single gene disorders.

<pubmed>20966459</pubmed> This article gives detailed laboratory instructions and guidelines for PGD procedures, which might be useful for the methodological part of the website

The following articles are about diseased cells/embryos derived from PGD procedures for further research: <pubmed>23242925</pubmed> <pubmed>22735930</pubmed>

Other articles <pubmed>21748341</pubmed>





--Mark Hill (talk) 17:45, 3 September 2015 (AEST) These are relevant to your project. I hope you get to use some in the final project. (5/5)

Lab 4 - Online Assessment

Fertilization Quiz

1 Which of the following statements about female and male gametogenesis is incorrect:

While male meiosis is completed in days or weeks, female meiosis is delayed for months or years.
Male meiosis produces four gametes, whereas female meiosis produces only two.
In oogenesis meiosis is initiated once in a finite cell population, while in spermatogenesis meiosis is continuously initiated in a stem cell population.
Male gamete differentiation occurs after meiosis ends, whereas in oogenesis this occurs earlier during the first meiotic prophase.

2 Where does fertilization usually occur:

Upper uterine cavity
First 1/3 of uterine tube
Last 1/3 of uterine tube
Lower uterine cavity

3 The acrosome reaction...:

Helps spermatozoa to penetrate the zona pellucida
Initiates ovulation
Allows spermatozoa movement by stimulating the mitochondria
Is responsible for ZP2 expression in the zona pellucida
all of the above

--Mark Hill (talk) 17:50, 3 September 2015 (AEST) Q1 and Q2 are too easy and require simple guesses. You have not explained why the other options are incorrect in your answer and could have linked to further resources. (8/10)

ANAT2341 Student 2015 Quiz Questions

Lab 5 - Online Assessment

Interferon Regulatory Factor-6 and Cleft Lip and/or Palate

Orofacial clefts such as cleft lip and/or palate are common structural birth defects with birth prevalence ranging from 1/500 to 1/2,000 in different populations. The failure in growth of the frontonasal prominence, the paired mandibular processes, the paired maxillary processes, and the medial and lateral nasal processes in week 4, together with the failure in fusion of the lateral nasal processes with the maxillary processes, and the medial nasal processes in week 6 and 7 results in orofacial clefting of the upper lip and/or primary palate. Clefts in the secondary palate may arise due to failure in several developmental steps after week 6, such as palatal shelves elevation or migration[4].Their complex etiology is not fully understood and both genetic and environmental causes have been established to be involved in cleft lip and/or palate development. Many aberrations in different genes have been found to correlate with cleft occurrence. One pivotal gene appears to be located on chromosome 1 at the 1q32 region, which encodes interferon regulatory factor-6 (IRF6)[5], an important member of the IRF family involved in oral and maxillofacial development[6].

Mutations in IRF6 were first identified in patients suffering from Van der Woude syndrome, who often display orofacial clefts in addition to other symptoms. In several subsequent research studies SNPs in IRF6 were also detected in some non-syndromic cleft lip and/or palate. Irregularities in IRF6, therefore, are considered risk factors for cleft lip and/or palate development[7]. The phenotypic heterogeneity of Van der Woude syndrome in comparison to non-syndromic cleft lip and/or palate is hypothesized to be caused by different types of mutations of IRF6 resulting in either a partially or fully nonfunctional protein[8]. In addition, the phenotype might be influenced by the site of mutation[9]. Mutations on a specific sequence variant about ten kb upstream of its transcription start site have been found to keep transcription factor AP-2α from binding and, therefore, influencing IRF6 expression[7].

Many animal models have given rise to several hypotheses about how IRF6 irregularities may affect development on a molecular level. For instance, IRF6 mutations caused a hyper-proliferative epidermis in mice. This will induce a failure of terminal differentiation in the respective epidermis and generate epithelial adhesions that can clog the oral cavity and create a cleft palate further on during development. IRF6 also has been identified as a key determinant of keratinocyte proliferation, oral periderm formation, and its spatio-temporal regulation. Additionally, IRF6 in interaction with other transcription factors has been studied widely. For example, p63 activates IRF6 transcription and is mutated in many malformation syndromes that display cleft lip and/or palate. This emphasizes IRF6's role in facial development and in the etiology of orofacial clefts[7]. Studies in zebrafish and frog embryos have provided information about the interaction between IRF6 and Grainyhead-like 3 (Grhl3), which has effects on regulation of the epidermal permeability barrier and on periderm differentiation. IRF6 seems to directly active Grhl3 expression by binding to its promotor. This Grhl3 promotor binding association has also been observed in humans[10]. Several other hypotheses of IRF6's involvement in the etiology of cleft lip and/or palate have been proposed, however, the precise mechanisms are not known and more research is needed to establish a full understanding[11].

--Mark Hill (talk) 14:20, 7 November 2015 (AEST) (5/5)

Lab 7 - Online Assessment

1. Identify and write a brief description of the findings of a recent research paper on development of one of the endocrine organs covered in today's practical.


This study tried to elucidate the pathways for β-cell generation and investigated in particular the potential functions of 17β-estradiol (E2) and oestrogen receptor α (ERα) signalling in the stimulation of β-cell generation during embryonic development and in injured adult pancreases. The study focused particularly on the transcription factor Neurogenin 3 (Ngn3), which is involved in the developmental program that generates embryonic β-cells. Mice studies with different knock-add-on ERα antagonists were performed to investigate oestrogen's and ER's roles in β-cell proliferation and differentiation. In addition, the ERα signalling was impaired chemically. The loss of ERα signalling results in decreased expression of Ngn3 in pancreatic progenitors. This consequently decreased amounts of progenitor cells and β-cell. It is hypothesized that the ERα signalling interferes with notch expression which counteracts the further differentiation of pancreatic progenitors. However, as low levels Ngn3 were present other molecular processes besides ERα seem to be involved in its regulation. An upregulation of ERα signalling was recorded adult mice severely injured pancreatic cells that recruited Ngn3-positive endocrine progenitor-like cells. The study, therefore, concludes with establishing the possibility of an existing relationship between ERα signalling and β-cell proliferation and differentiation. However, more research is needed to fully establish a causal relationship[12].

2. Identify the embryonic layers and tissues that contribute to the developing teeth.

Teeth development is derived from ectoderm of the first pharyngeal arch, mesoderm, and neural crest (ectomesenchymal cells) contributions and is based on these epithelial/mesenchymal interactions. In specific odontoblasts and ameloblasts are involved. The former are neural crest-derived mesenchymal cells that differentiate under the influence of enamel epithelium. They also produce predentin and eventually calcify in order to form dentin. The latter are inner enamel epitherlial cells that produce enamel. In addition periodontal ligaments, which coat the tooth root with cementum, are needed to hold the tooth in the bone socket (alveolar bone), while the teeth are growing in the ossifying jaw. The teeth are not directly anchored into the alveolar bones but rather held in place by this specialized connective tissue structure. It has additional functions, such as transmitting chewing forces and contains collagen fiber bundles that are known as "Sharpey's fibres".

--Mark Hill (talk) 14:20, 7 November 2015 (AEST) (5/5)

Lab 9 - Online Assessment/ Peer Reviews

Group 1

The group project page ‘Three Persons Embryos’ covers a lot of aspects of this topic. The page incorporates the example of Alana Saarinen. This ‘personal’ case study makes the topic interesting and makes people want to learn more about it. The video is a good way of introducing the reader to the page, but maybe a summary would be useful for people who cannot/ do not want to access the video. Overall, the headings are in a logical order and the subheadings are useful. However, it might make it easier to follow the page if the subheadings for each of the techniques were unified, e.g. “Procedure” “Animal Models” “Current Research” for each procedure. Also, the heading “Benefits” appears to be rather a description of the indications for the procedure, a renaming of this heading might clarify this.

Many key points relating to the topic have already been mentioned, but expanding further on the majority of headings and subheadings would be useful. Several useful pubmed articles are mentioned where the text is only short, so only their summary is missing. The referencing appears to be correct most of the time, however, there are several instances where one paper was referenced several times individually. Check to learn how to avoid this.

In addition, the three uploaded images need to be double checked for their copyrights, as they do not clearly state an allowance for the reuse of their content. In case, they cannot be reused, the images could be self-drawn. The timelines are interesting and necessary components of the project. They could also be displayed as actual timeline-graphs. This would add to the amount of graphs used and illustrate the content nicely. Overall it might be useful to check for spelling and grammar mistakes before the final deadline.

Group 2

The contents of the website about OHSS appear to be very well researched and the key points of the topic are clearly described. The headings and subheadings are in a logical and complete order. However, it might be useful to add a new heading labelled “Current Research” about e.g. the pathophysiology, treatments, diagnosis, etc. It might be that this will be included in the “animal models” section, which is still incomplete. Upon completion of the last sections it might be useful to round the website off with a conclusion.

The website so far has only one image. However, the image is self-drawn and very clear. The image is useful, readable, and makes the understanding of the pathophysiological processes easier. It might be good to consider adding more graphs and images to the website. Useful images could display the involved anatomical structures, the symptoms, and the diagnostic procedures (Ultra-sound showing OHSS).

The used references are all very recent, which strengthens the credibility of the website. However, sometimes sentences or paragraphs are not cited at all. Thus, lacking citations should be added. The glossary is a nice addition to the website but still needs to be completed (VEGF, LHR, BMP-15, etc.)

It might be beneficial if the link to ART were explained more clearly. As the general frame of the project is ART, including OHSS’ implications on the procedure, causative role, etc. would elucidate that link.

Group 3

The website appears very well researched and covers all important points relating to the topic PCOS. Thus, it might be reasonable to change the topic of the entire page to PCOS and not female infertility, as that does not seem to be the real focus of the website. Otherwise, this might seem a little confusing to readers. In addition, maybe try to relate the topic to ART.

The website has many useful images, diagrams, and tables, which are useful for the understanding and illustration of the written content. The self-drawn image is a very good anatomical illustration. Nevertheless, there are many references and a lot of information to be found on the website and inclusions such as the animal models, etc. really go in depth of the topic.

The headings and subheadings are all in a logical order and cover all relevant aspects of the topic. Content wise, it might be good to look into the causes again and see if there is any additional information on the environmental factors, obesity and diet, and medication, as these are comparatively short. Also, the definition heading could probably be labelled epidemiology.

The display of the treatment options is really great and highlights all important information. In general the page does very well on highlighting important features (purple highlights). Maybe add some more of these to be more consistent.

Overall, the referencing is very good. Only the environmental factor subheading has no reference yet and the referencing of the Ultrasound of Polycystic Ovaries image is not according to Mark’s guidelines (only the PMID is shown, not the actual name of the paper,authors).

Group 4

The website is very well organized and contains a lot of researched information and content. The introduction gives a short and precise definition of male infertility and let’s the reader know what to expect from the website. Overall the order of the headings and subheadings is very logical. To start off with explaining the healthy male physiology is a smart way to allow readers with less knowledge about the topic to understand the website. The blue theme of the tables is very coherent and the recurrent incorporation of the tables is alternative to plain text. The “treatment” heading has a lot of information ranging from traditional to Western medicine, which shows that the topic was very well researched.

In addition, it is good to see that at the end of the treatment section (upon completion) the page will give a direct link to ART. The referencing of the website and the images is correct. Here and there are some paragraphs or tables that are not referenced yet (Classifications of Valsalva manoeuvre, Intro. of “Diagnosis”). The amount of references is extensive showing again that the topic has been researched quite well. The images are very useful, for understanding the content of the website. Particularly the diagrams explaining the Varicocele and the mechanisms of Lycopene Treatment clarify a lot. The timeline of the development of gonadotropin preparations is also a great way to incorporate bits of the history of your topic without spending too much text on it.

It might be useful to add another heading about current research and a glossary to help readers to understand the website. It is really great that you added a video to your website, particularly in the beginning as it will introduce to topic to readers and ease them into the topic. However, it might be reasonable to look for a video that only focuses on male infertility. If that cannot be found it would not be too bad, as the video is still relevant to the topic overall. Moreover, try to add a self-drawn image, maybe in the treatments part for the surgical interventions?.

Group 5

The website contains a lot of information and covers all key points relevant to the topic. It shows that the topic of oncofertility and cancer in general has been researched very well. The addition of many pictures, graphs and tables is useful for understanding the written contents. The videos are a great way to bring some variety into the website and illustrate cancer and chemotherapy well. It might be nice to look for videos for the oncofertility section. Making several keywords bold is a good way to stress their importance and helps readers to orientate around your website. The timeline of oncofertility is very extensive and the table is a great way of presenting this information.

Overall, the structure of the headings is a bit confusing. It might be more logical to place the “chemotherapy” section before the “infertility” heading. Moreover the “infertility” section appears to be rather about the causes of infertility instead of describing what infertility is, so renaming might useful. The referencing overall is good, however, here and there some references are missing, for instance in “Bone marrow or stem cell transplant” or “How Does Chemotherapy Work?”. Additionally, some references were used more than once using the wrong code, which makes them show up several times in the reference list. Check to learn how to avoid this. The image ‘IVF flow chart’ does not include the copyright information.Overall, double check the copyright and referencing of the images under the chemotherapy sections.

As for the tables comparing fertility preservation in men and women, the presentation in two identical tables is very helpful. However, adding some more information for the men one or changing something of the layout of the table would make the table look less incomplete. The way it is now makes it look a little ‘neglected’. It might be interesting to add a “current research” section and a glossary to clarify terms.

--Mark Hill (talk) 14:20, 7 November 2015 (AEST) These are well thought out peer assessments and it is obvious you have spent some time preparing this feedback. (17/20)

Lab 10 - Online Assessment

Retinal Pigment Epithelium

Link to permalink image: Retinal Pigment Epithelium

The Retinal Pigment Epithelium (RPE) is a complex differentiation of the retina,is generated from the optic neuroepithelium, and is structurally made up of cuboidal cells and multiple villi on its apical side. Its lateral sides are joined together by gap junctions and adherens and the RPE's basal side is in contact with Bruch's membrane. It lies between the neuronal retina and the choroid. The section shows that in the embryo the pigmented retina is still separated by a space from the neuronal retina. This space will be decreased in the adult and closely appose the two to each other.

Embryology Link Vision - Retina Development#Retinal Pigment Epithelium

--Mark Hill (talk) 14:25, 7 November 2015 (AEST) Very good. (5/5)


PMID 26193891 PMID 26161332 PMID 25935518 PMID 26168107 PMID 22723007 PMID 24515905 PMID 20966459 PMID 23242925 PMID 22735930 PMID 15185170 PMID 21331089 PMID 23940636 PMID 22438645 PMID 23949966 PMID 24124047 PMID 22931925 PMID 26332872 PMID 26015547

--Mark Hill (talk) 14:45, 7 November 2015 (AEST) Stem Cell Presentation (17/20)

--Mark Hill (talk) 08:40, 1 November 2015 (AEDT) Catei submitted (5)

  1. <pubmed>26193891</pubmed>
  2. <pubmed>26161332</pubmed>
  3. <pubmed>25935518</pubmed>
  4. <pubmed>24124047</pubmed>
  5. <pubmed>15185170</pubmed>
  6. <pubmed>23940636</pubmed>
  7. 7.0 7.1 7.2 <pubmed>21331089</pubmed>
  8. <pubmed>22438645</pubmed>
  9. <pubmed>23949966</pubmed>
  10. <pubmed>22931925</pubmed>
  11. <pubmed>26332872</pubmed>
  12. <pubmed>26015547</pubmed>