The purpose of this experiment was to examine the cytoskeleton, its role in cellular shape and adhesion; as well as identify the molecules necessary for cytoskeleton function. There were two experiments preformed, one involving RAW cells and the second involving rabbit skeletal muscle cells. The first experiment required the RAW cells to be exposed to different concentrations of the drug cytochalasin D. This particular drug inhibits the polymerization actin, a protein microfilament involved in the cytoskeleton. It provides shape to cell and is involved in cell adhesion. After incubation of the experimental cultures they were examined for changes in cell structure in comparison to the control cells with no exposure to the drug. The latter experiment …show more content…
involved exposing the skeletal muscle cells to solutions with varying solute content. Using a light compound microscope, the striations in the muscle fibres were examined to see if solute content would effect the cells ability to contract (Heidcamp et al., 2017). Part B: 1. The average number of adhered cells is 2. 3. According to the data collected there were no concentrations of Cytochalasin D that lowered cell adhesion to more than 50% relative to the control cells which on average was 6 cells. The highest adhesion rate was observed with the lowest concentration of Cytochalasin D added and lowered with increasing concentrations of Cytochalasin D but never below the average of the control. 4. Based on the data the Cytochalasin D concentrations may be grouped into “cells adhered and stretched” versus “cells that adhered but stayed rounded”. Concentrations of 5nM to 10 nM showed cells which were adhered and stretched. Concentrations of 100 nM to 200 nM showed cells that adhered but stayed rounded. The concentration of 50 nM of Cytochalasin D showed approximately the same number of rounded cells and stretched cells, being a balance between the two. 5. The purpose of adding DMSO to the control (vehicle) tubes is to prevent cells from being damaged. DMSO is involved in the freezing of cells and reduces formation of ice crystals in a cell culture during the freezing process (Pubchem, 2017). Without DMSO to reduced crystal formation the cells would become damaged or destroyed and ultimately not viable for data collection. 6. Based on the observations and measurements collected in exercise 3.2, muscle contraction requires , and molecules to be in the extracellular fluid volume in order to happen. The data showed that only solution 2 with all three molecules caused a decrease in length of striation. A decrease in this length would translate to the muscle fibre contractions. Part C: Research 1. Fibronectin is a large glycoprotein found in the extracellular matrix in all vertebrates (Alberts et al., 2008). It is involved in cell adhesion and works by bonding to protein receptors on the surface of a cell. F-selectin is a selectin protein predicted to be involved in cell adhesion to fibronectin. All selectin need ions to function (Alberts et al., 2008). A possible experiment to show that F-selectin can mediate cell adhesion to fibronectin can be the following: Create six cultures of the cell line MD1 suspended in a solution of DMSO and , six cultures of the cell line MD1FS suspended in a solution of DMSO and , create one culture of the original cell line suspended in DMSO and , and create one culture of the original cell line only suspended in DMSO.
The culture of the original cell line suspended in DMSO and will act as a control for the first 12 cultures mentioned. The culture of the original cell line only suspended in DMSO will act as a control from the original with DMSO and to see if the added ions affect cell adhesion. The multiple test cultures are to insure consistency in collected …show more content…
data. The 16 cultures will all be incubated, aspirated of the solutions, and then fixed with a solution of paraformaldehyde, the cells will be incubated once more, aspirated of the solution and then mounted to slides to be observed under a microscope. The expected outcomes for these cell cultures is that the cell line of MD1 would show the most number of adhered cells while the cell line MD1FS would show much less or no adhered cells.
This is expected because if F-selectin is responsible for cell adhesion to fibronectin surfaces, F-selectin would need to be express to observe adhesion. Therefore, the cell line that does express F-selection would show much more cell adhesion. The original cell line suspended in DMSO and would show results much similar to the MD1 line and the original cell line only suspended in DMSO would show very littler or no adhered cells because of the lack of essential ions in the extracellular matrix needed for selectin to work if selectin does mediate cell adhesion to
fibronectin. References Dimethyl sulfoxide | (CH3)2SO - PubChem. (n.d.). Retrieved March 16, 2017, from https://pubchem.ncbi.nlm.nih.gov/compound/dimethyl_sulfoxide#section=Top Johnson, A., Lewis, J., Raff, M., Roberts, K., & Walter, P. (2008). Chapter 19: Cell Junctions, Cell Adhesion, and the Extracellular Matrix. In B. Alberts (Author), Molecular Biology of the Cell (5th ed., pp. 1145-1146, 1191-1193). New York, NY: Garland Science. Heidcampl, W. (2017). Cell Biology II Laboratory Manual (C. Antonescu, R. Botelho, & L. Victorio - Walz, Eds.).
In the case of temperatures the cultures were incubated at each determined temperature. For the UV radiation, cells were exposed to UV light for 10 seconds and then grown in 30oC. For the EtBr treatment, 50ul of EtBr was added to the growth medium and cells were incubated at 30oC. In the case of sunlight exposure, cells were exposed to sunlight directly and grown at room temperature
...et light. If the LAA plate glows green under exposure to ultraviolet light, then we can conclude that our unknown insert piece of DNA would be the kan gene. If it does not glow green under exposure to ultraviolet light, then then we streak the colony from our LAA plate onto the LAC plate using a sterile glass spreader. When the LAC plate is dray, we place it upside down in the microfuge rack so that it can be incubated at 37 ºC. Incubation at 37 ºC will allow the transformed bacterial cells to grow. If we see bacterial growth on the LA plate containing chloramphenicol, we can conclude that our unknown insert piece of DNA would be the cat gene, since the cat gene is resistant to chloramphenicol. Afterwards, we then grab the microfuge tube labeled NP and repeat the aforementioned steps shown above pertaining to the LA plates. This would be considered our control.
...st the sacrolemma will depolarized, thus activation potentials along the T-tubules. This signal will transmit from along the T-tubules to sarcroplasmic reticulum's terminal sacs. Next, sarcoplasmic reticulum will release the calcium into the sarcroplasm leading to the next second event called contraction. The released calcium ions will now bind to troponin. This will cause the inhibition of actin and mysoin interaction to be released. The crossbridge of myosin filaments that are attached to the actin filaments, thus causing tension to be exerted and the muscles will shorten by sliding filament mechanism. The last event is called Relaxation. After the sliding of the filament mechanism, the calcium will be slowly pumped back into the scaroplasmic reticulum. The crossbridges will detach from the filaments. The inhibition of the actin and myosin will go back to normal.
No obvious boundaries are observed between the two distinct materials, tendon and bone. Type I collagen and tenocytes are highly aligned in tendon. In uncalcified fibrocartilage, where collagen type II is of great content, along with rich type III collagen and small amount of type X collagen, decorin, and aggrecan. Similarly, with a great amount of type II collagen, the mineralized cartilage presents significant amounts of collagen type X and scarce levels of aggrecan. Note that the collagen fibers are highly aligned in the direction of tensile force in tendon but less oriented in the insertion site (Figure 2).[4, 19] Additionally, the insertion site possesses a transitional decrease in tissue organization while an increase in mineral content.[4] The complex collagen and mineralization content in this region lead the repair and rehabilitation of tendon-to-bone insertion site more
Obesity a risk factor in which excess body fat accumulates and can have negative effects on your health. Here we identify how the hormone insulin reacts in 3T3-L1 fibroblasts and its role on adipogenesis. Adipogenesis is the development of fat cells from pre adipocytes. Insulin is an important factor in the differentiation of 3T3-L1 pre adipocytes to mature adipocytes. Oil Red O (ORO) is used to demonstrate the presence of lipids in each different treatment. A spectrophotometer is used to get the optical density of liquid at the different insulin concentrations. One factor CREB is revealed from preadipocytes to mature adipocytes. By demonstrating how insulin triggers transcription factors. When cells are insulin induced CREB is activated in differentiation. Insulin increased the rate of differentiation and the amassing of triglycerides in 3T3-L1 cells . Insulin was able to induce adipogenesis by observing cell morphology and optical density of liquid from ORO stain. Insulin at 1 µg/ml had the optimal rate of differentiation compared to the other insulin concentrations. Morphology of cells changed significantly from Day 0 to Day 7 at 1 µg/ml and appeared larger and
Repair after a muscle is damaged happens through the division of certain cells who then fuse to existing, undamaged muscle fibers to correct the damage. Different muscle types take different amounts of time to heal and regenerate after it has been damaged. Smooth muscle cells can regenerate with the greatest capacity due to their ability to divide and create many more cells to help out. While cardiac muscle cells hardly regenerate at all due to the lack of specialized cells that aid in repair and regeneration. In skeletal muscle, satellite cells aid in helping restoration after injury. Along with muscles, tendons are very important structures within the human body, and they to can be damaged. However, tendon repair involves fibroblast cells cross-linking collagen fibers that aid in not only reinforcing structural support, but also mechanical support as well (“Understanding Tendon Injury,” 2005). While quite different from muscle repair, tendon repair involves the similarity of reestablishing d...
The first step to the research is to be able to have large enough cell colonies (a cell colony is where a group of cells are grown to be researched on) but even cell colonies can
The experiment that tested the contractile level of muscle in various solutions used a muscle fiber from rabbit’s muscle. One fiber was detached, put under microscope, and submerged first under ATP and salt solution (KCl and MgCl2), then ATP only solution, and lastly salt only solution [2]. The fiber’s level of contraction was measured in micrometers. Muscle contractile strength and number motor units employed at various force lev...
The sarcomere is found in structures called myofibrils which make up skeletal muscle fibres. Within the sarcomere there are various different proteins. One of the most significant, myosin is found in the thick filaments of the sarcomere. Although both cells contain myosin, it is important to highlight that smooth muscle cells contain a much lower percentage of myosin compared to skeletal muscle cells. Despite this, myosin filaments in smooth muscle cells bind to actin filaments in a manner similar to that in skeletal muscle cells; although there are some differences. For instance, myosin filaments in smooth muscle cells are saturated with myosin heads so that myosin can glide over bound actin filaments over longer distances, enabling smooth muscle cells to stretch further, whilst in skeleta...
LAB REPORT 1st Experiment done in class Introduction: Agarose gel electrophoresis separates molecules by their size, shape, and charge. Biomolecules such as DNA, RNA and proteins, are some examples. Buffered samples such as glycerol and glucose are loaded into a gel. An electrical current is placed across the gel.
The cytoskeleton is made up of three different types of filaments, actin filaments, intermediate filaments and microtubules. Actin filaments are the thinnest, they are also known as microfilaments. They create a band under the plasma membrane, this gives strength to the cell and links transmembrane proteins such as cell surface receptors to cytoplasmic proteins. Intermediate filaments include keratins, lamins, neurofilaments and vimentins. Keratins form hooves, horns and hair and are found in epithelial cells. Lamins form a type of mesh that ‘stabilizes the inner membrane of the nuclear envelope’ (Biology Pages). Neurofilaments bring strength to the axons of neurons and vimentins provide mechanical support to cells – particularly muscles. The cytoskeleton is also involved in cell
In the nursing profession, strategic thinking, effective decision making and delegation is important. With this, the goal of providing quality care is key and can be accomplished through a theory called servant leadership. In this style of leadership and management, the entire team has input into decision making based on the organization’s values and ideals. Servant leaders create devoted followers in response to the positive attention they give (Nursing Community Journal, 2015).
The Cell, the fundamental structural unit of all living organisms. Some cells are complete organisms, such as the unicellular bacteria and protozoa, others, such as nerve, liver, and muscle cells, are specialized components of multicellular organisms. In another words, without cells we wouldn’t be able to live or function correctly. There are Animal Cells and Plant Cells. In Biology class the other day we studied the Animal Cell. We were split into groups of our own and we each picked a different animal cell slide to observe. My group chose the slide,'; Smeared Frog Blood ';.
Tissue culture allows for the clonal propagation of plant (production of multiple copies of the same genotype).