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Attendance

--Z3330409 (talk) 17:05, 21 March 2013 (EST)
--Z3330409 (talk) 15:11, 28 March 2013 (EST)
--Z3330409 (talk) 15:07, 11 April 2013 (EST)
--Z3330409 (talk) 15:16, 18 April 2013 (EST)
--Z3330409 (talk) 16:46, 9 May 2013 (EST)
--Z3330409 (talk) 16:44, 16 May 2013 (EST)

Individual Assessments

Lab 1

Inferred lifecycle of eukaryote ancestor.jpg

Figure 2. Inferred life cycle and high degree of organellar complexity of the last common ancestor of all extant eukaryotes. Origin of the cell nucleus, mitosis and sex: roles of intracellular coevolution Thomas Cavalier-Smith http://www.biology-direct.com/content/5/1/7

© 2010 Cavalier-Smith; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Lab 2

The use of confocal microscopy in observing melanin and the skin

The human skin is a complex organ in the way that it protects the rest of the body from light and UV penetration. Although its has been determined that “skin optics respect the rules of absorption, reflection and scattering” (Corcuff, P et al 2001), the skin still has features that may prevent direct penetration of light. These include
• The stratified, layered nature of the epidermis
• Biochemical composition of both the epidermis and dermis
• Melanin within the epidermis and haemoglobin from blood in the dermis.
For these reasons, in vivo observation of the skin in its physiology and responses to stimuli has always been somewhat of a challenge. What Corcuff, P et al experimented with was a method of observing the skin of the forearm before sun exposure and for a month afterward using confocal microscopy techniques. Observations were made in regards to melanasome (melanin-containing organelle) caps within basal keratinocytes (skin cells in the deep epidermis). The use of in vivo confocal microscopy also “affords new insight to the role of melanin and its gradual migration after sun exposure”

In vivo confocal microscopy involved reflected singals being used to create an image. Contrast was improved due to absorption and scattering (minimal loss of photons). Another feature that was regarded was realtime imaging to prevent blurring from motion caused by blood pulses and involuntary movement.


• Skin optics revisited by in vivo confocal microscopy: Melanin and sun exposure
PIERRE CORCUFF, CIdLINE CHAUSSEPIED, GENEVIEVE MADRY, and CHRISTOPHE HADJUR, L'Ordal
Journal of Cosmetic Science, 52, 91-102 (March/April 2001)

http://journal.scconline.org/pdf/cc2001/cc052n02/p00091-p00102.pdf

--Z3330409 (talk) 18:02, 27 March 2013 (EST)


--Mark Hill (talk) 11:29, 11 April 2013 (EST) You have identified a research article using in vivo confocal imaging of the skin and this meets the criteria for the microscopy. The paper though is from 2001 (12 years ago) and I do not think this meets the "recent research article" criteria. Furthermore you have not used the Pubmed formatting as described in the in-class tutorial and as shown below, see also Project Referencing. Both these factors affected your final mark.

<pubmed>11382846</pubmed>



Lab 3

1) Fragmentation and partitioning of the Golgi apparatus during mitosis in HeLa cells

HeLa cells are a type of human cell used in scientific research obtained from cervical cancer cells in 1951. These cells were used to observe the activity of the Golgi apparatus when they divide. It has been concluded that animal cells contain only one copy of the Golgi apparatus and they are unable to be synthesised de novo. The Golgi must divide when the cell divides and involves breaking down into fragments. By using osmium to stain the Golgi, Lococq and Warren were able to demonstrate the division of the organelle through twisting and turning in a ribbon form.

Lucocq JM, Warren G (1987) Fragmentation and partitioning of the Golgi apparatus during mitosis in HeLa cells. The EMBO Jounal 6(11)



2) Active ADP-ribosylation Factor-1 (ARF1) is required for Mitotic Golgi Fragmentation.

Although it has been established that animal cell Golgi apparatuses undergo division and disassembly during mitosis, the mechanism for the way this occurs is still not clear. Xiang Y et. al. investigated how ARF1 may dictate the division of the Golgi. ARF1 is a GTPase that is necessary for the forming of vesicles from the Golgi and is associated with Golgi vesicles created in vitro as well as in mitotic cells. When the Golgi was treated with ARF1, it was converted to vesicles. However depletion of ARF1 meant the Golgi failed to fragment.

Xiang Y et. al. (2007) Active ADP-ribosylation Factor-1 (ARF1) is required for Mitotic Golgi Fragmentation. Journal Of Biological Chemistry 282(30)



3) The Golgi-associated protein GRASP65 regulates spindle dynamics and is essential for cell division

It is found that that the protein known as GRASP65 (a Golgi-associated protein) is significant in the in Golgi fragmentation during mitosis. GRASP65 is required for the stacking of Golgi cisternae in vitro and is phospohrylated during mitosis to cause unstacking during mitosis. The study showed that depletion of GRASP65 did not affect the stacking of cisternae however it did afect the organisation of the mitotic spindle. This ultimately lead to cell death.

Sutterlin C, Polishchuk R, Pecot M, Malhotra V (2005) The Golgi-associated protein GRASP65 regulates spindle dynamics and is essential for cell division. Molecular Biology of the Cell Vol 16



4) The Golgi and Endoplasmic reticulum remain independent during mitosis in HeLa cells

The Golgi apparatus must undergo partitioning and dissasembly prior to mitosis. And although it may not require molecular machinery to dissasemble/ reassemble, it requires a number of specific cell-cycle regulated activities. The same can be said about the Endoplasmic reticulum which is a large tubular network that involves the nuclear membrane. In this study, Jesch SA et. al. conducted a number of assays to observe wether the Golgi apparatus and ER are dependent or independent of one another during mitosis

Jesch SA, Linstedt AD (1998) The Golgi and Endoplasmic reticulum remain independent during mitosis in HeLa cells. Molecular Biology of the Cell 9(3).



Stained Golgi.png

--Mark Hill (talk) 12:12, 3 May 2013 (EST) I cannot see appropriate information associated with this image from EMBO J that allows you to reuse here. Unless you provide this information I will be deleting this image.

Lab 4

Integrin α1 Antibody (A-9): sc-271034

-Monoclonal antibody
-Isotype - IgG (made of 4 peptide chains - 2 light, 2 heavy)
-Raised in mice, used within mice
-Provided at 200µg/ml
-Recommended for detection of Integrin α1 of human and rat origin by Western Blot, IP, IF and ELISA
-


http://www.scbt.com/datasheet-271034-integrin-alpha1-a-9-antibody.html
http://datasheets.scbt.com/sc-271034.pdf


Lab 7

Graphs of cell phenotype.png