From CellBiology

--Z3336156 15:13, 8 March 2012 (EST)

Lab Attendance

--Z3336156 15:29, 8 March 2012 (EST) --Z3336156 14:11, 15 March 2012 (EST) --Z3336156 14:08, 22 March 2012 (EST) --Z3336156 14:09, 29 March 2012 (EST) --Z3336156 14:07, 5 April 2012 (EST) --Z3336156 14:07, 19 April 2012 (EST) --Z3336156 14:19, 26 April 2012 (EST) --Z3336156 14:05, 3 May 2012 (EST) --Z3336156 13:59, 10 May 2012 (EST)

Lab 1 (week2) JOVE Lecture 2

Cell Nucleus

Hi guys!

Lab 2 (week3)

Collapsing Actin Mesh.jpg


The exact location of nucleiods which contain mitochondrial DNA has been vaguely described and was a mystery due to the lack of available technology, which could measure precise locations of minute organelles molecules, that lie inside organelles. However, since the discovery of superresolution microscopy scientists were able to study further the unsolved questions of nucleiods found in the mitochondria. Brown et. al were able to discover the shape, size and relative locations of nucleoids with superb imaging. Three dimensional images showed that mitochondrial DNA are found in ellipsoidal nucleoids and is arranged in an extremely condensed fashion.

Reference: <pubmed> 22006021</pubmed>

--Mark Hill 12:50, 20 March 2012 (EST) Good explanation of the study findings.

Lab 3 (week4)


Journal Articles About VEGF:

1) The journal article was researched about the effect of Hydrogen Sulfide to VEGF during myocardial infarction. VEGF binds to the tyrosine kinase and fms-like tyrosine kinase receptor that can up-regulate or down regulate vascular organisation. The research showed that mice treated with hydrogen sulfide after an induced myocardial infarct had higher levels of VEGF and the 2 receptors mentioned earlier that resulted in a better recovery after the injury.

Reference: <pubmed>22419888</pubmed>

2) In the research conducted by Tororovic et al. they studied the correlation of the presence of VEFG and the proliferation of leukemic blast cells and microvessel density found in acute lymphoblastic leukemia(ALL) patients. The study found that increased VEGF leads to higher levels of leukemic blast cells and microvessel density count. This finding can be used to seek new methods in combating ALL.

Reference: <pubmed>22417860</pubmed>

3) The article talks about the effect of VEGF which binds on VEGFR-1(vascular endothelial growth factor-1) and VEGFR-2(vascular endothelial growth factor 2) that affects the astrocytes in terms of gap junctional intercellular communication, cell proliferation and motility of astrocytes. The experiment was conducted through the nurturing of astrocytes in VEFG rich solutions, from here, it was discovered that VEGF promotes gap junctional intercellular communication, increases cell mitosis and enhances the motility of astrocytes through the VEGFR-2.

Reference: <pubmed>22431192</pubmed>

4) VEGF plays major role in the progression of osteoarthritis thought the proliferation of blood vessels in the subchondral growth plate. It was hypothesized that VEGF plays a role in the early stage of osteoarthritis as a marker for diagnosis that can be used as a marker for diagnosis and was proven to be correct.

Reference: <pubmed>22429866</pubmed>

SDS for Methanol:

Properties: state: liquid appearance: colourless odour: alcohol-like, weak odour solubility in water: miscible

Hazards: POISON. Ingestion can be fatal and cause colour blindness. Vapour is flammable and still harmful. Harmful if swallowed, inhaled, or absorbed through the skin. Eye, skin, and respiratory tract irritant.

Potential Health Effects Eye: May cause painful sensitization to light. Methanol is a mild to moderate eye irritant. Inhalation, ingestion or skin absorption of methanol can cause significant disturbance in vision, including blindness.

Skin: Causes moderate skin irritation. May be absorbed through the skin in harmful amounts. Prolonged and or repeated contact may cause defatting of skin and dermatitis. Methanol can be absorbed through the skin, producing systemic effects that include visual disturbances.

Ingestion: May be fatal or cause blindness if swallowed. Aspiration hazard. Cannot be made nonpoisonous. May cause gastrointestinal irritation with nausea, vomiting and diarrhea. May cause systematic toxicity with acidosis. May cause central nervous system depression, characterized by excitement, followed by headache, dizziness, drowsiness, and nausea. Advanced stages may cause collapse, unconsciousness,


Lab 4 (week5)

Musashi Protein <pubmed>16717192</pubmed> Musashi is responsible for the balance between germ-line stem cell renewal and differentiation. Its function in stem cells is for maintenance of stem cell identity. The paper also found that Musashi plays various roles in the different stages of germ cell differentiation in the male meiosis(spermatogenesis).

Primary Musashi Antibody: Musashi-1 RBT REC OLIGO AB

Musashi-1 ABfinity™ Musashi-1 SDS for RBT REC OLIGO AB

Antibody:Musashi-1 RBT REC OLIGO AB

Host: Rabbits

Price: $280.00 (usd) /100 µG

Clonality: polyclonal (oligoclonal)

Secondary Musashi Antibody:

Anti-antibody: Alexa Fluor® 488 Goat Anti-Rabbit IgG (H+L) *2 mg⁄mL*

Host: Goat

Reactivity: Rabbit

Lab 5 (week6)

Image Relating to Topic

Alexa Fluor® 488 Goat Anti-Rabbit IgG (H+L) *2 mg⁄mL*

MSDS for Alexa Fluor® 488 Goat Anti-Rabbit IgG (H+L) *2 mg⁄mL*

VEGF signalling pathway .gif

Reference: <pubmed> 11700390 </pubmed>

Lab 6 (week7)

Tm4-Control Graph.png

Do you see a difference in phenotype morphology between Tm4 over –expressing cells with the control group?

Fan– the control group had almost four times the number of cell count as compared to Tm4 over-expressing cells

Broken Fan – the Tm4 over-expressing cells showed a 65% reduction in cell count in comparison with the control group

Stumped - the Tm4 over-expressing cells showed significant reduction of 30% in cell count in comparison with the control group

Pronged - the Tm4 over-expressing cells showed a great increase of 320% in cell count in comparison with the control group

Stringed - the Tm4 over-expressing cells showed an immense increase of 250% in cell count in comparison with the control group

Pygnotic - the Tm4 over-expressing cells was only 17% of the total number of control group cell count

Comparison of Tm4 over-expression and the control group

Tm4 Group:

-most of Tm4-overexpressers had a stringed or pronged phenotype

-there were few stumped and broken fans and very less fan

-there were just over a couple of cells with pygnotic phenotype

-the cells in this group were brighter and more fluorescent

-cells had more branching and longer processes

Control Group:

-most of the cells had a broken fan and stumped phenotype

-there were several fan, pronged, stringed and pygnotic cells of about the same amount

-cells had lesser branching and shorter processes

-the cells were not as fluorescent as compared to Tm4 over-expressers

How could Tm4 over-expression lead to this difference?

Tropomyson is an actin binding protein that affects the activity of actin. From the lab, it was found that Tm4 over-expression caused and increase in number of branching and length of processes in B35 cell phenotype. High levels of tropomyosin induce actin activity and prevent its disassembly. Further more, it increases the availability of actin to other actin related proteins leading to more developed filaments.[1] The differences seen from the two groups were caused by the over-expression of Tm4 in B35 neuroblastoma cells. Tm4 is found to increase the interaction of actin filaments that affected the cellular phenotype, for example, longer axons and dentries on neurons. Hence, over-expressers B35 cells had longer and more processes and branching. [2]

[1] <pubmed>19209825</pubmed>

[2] <pubmed> 7876361 </pubmed>

Lab 7

Contributions for group project:

Normal Function

Vascular Endothelial Growth Factors (VEGF) bind with the Vascular Endothelial Growth Factor Receptor (VEGFR) which initiate a cascade of signal resulting in vasculogenesis, angiogenesis and lymphangiogenesis in cells.

Several factors causes the production of VEGF namely:


All cells need oxygen to survive. However, in some cases not enough oxygen is supplied to tissues from various reasons, decrease in inspired oxygen from places of high altitude and preterm birth. Hypoxic cells, cells without adequate supply of oxygen can stimulate the production of VEGF. When cells are lacking oxygen, it produces Hypoxia-Inducible Factor (HIF) a transcription factor that stimulates the release of VEGF. The free VEGF then binds to VEGFR on cell membranes of endothelial cells causing angiogenesis and vasculogenesis as a cellular response.


An oncogene is a gene that can latently cause cancer. Oncogenes can be observed in high levels in tumour cells. Cell death or apoptosis is a normal process of a cell cycle, however, oncogenes suppresses this mechanism; hence, leading to further proliferation of mutated cells. Some of the manifestation of oncogenes is the error in protein structure that affects the level of enzyme activity and cell regulation. In cells with high levels of oncogenes, levels of kinase production are altered. Kinases are enzymes that add a phosphate group to different proteins which acts as an on and off switch as well as receptor kinases adding phosphate group to receptor proteins found in the surface of cell membrane to send out signal from the environment to the inside of the cell. Since VEGFRs are tyrosine kinase receptors, an alteration in the signaling pathway can cause cancer by switching on the receptor in spite having no signals from outside the cell.


-Angiotensin II

-Thyroid Stimulating Hormone (TSH)

-Corticotropin-Releasing Hormone

---Adrenocorticotropic Hormone


Other growth factors and cytokines

VEGF are also produced as a response to other growth factors and cytokines. Cox-2, cyclooxygenase-2 induces the production of VEGF. This correlation has been found with the study of tumors. PMID:11326316.