Friday, March 6, 2020

Chlamydomonas Lab Report Essays

Chlamydomonas Lab Report Essays Chlamydomonas Lab Report Paper Chlamydomonas Lab Report Paper Essay Topic: Whiplash The power stroke consists of beating stiffly in one direction, while during the recovery stroke the cilium is pulled back in floppy fashion close to the cell surface, thereby offering little resistance. Once in position, it straightens out and the stiff power stroke begins again. Human cilia or flagella are found in traditionally in three locations of the human body. In men, sperm is composed of a head and a tail, a tail that functions as flagella. In women, the fallopian tubes are lined with cilia projections, which help carry eggs and what creates resistance for sperm entering the vagina for fertilization. Both men and women (and all other mammals) have cilia in two other places in their bodies. One location is somewhat familiar: cilia coat the trachea and bronchial tubes leading down into the lungs. Cilia are also present in the brain and spinal cord. Cilia line the canals and ventricles, the empty spaces in the brain. The cerebral fluid in the brain and nervous system is kept in motion by the numerous cilia projections. Human cilia is very useful for multiple processes in the body, but when cilia is damaged, certain processes will not occur and defects will arise due to the lack of recesses. Cilia defects occur often in humans due to the defects in the cilia projections around the body. Cilia can suffer from genetic disorders, which are called caliphates. Caliphates usually involve proteins that localize to the primary cilia (the sensory organelles), which can cause genetic disorders such as primary cilia disease duskiness (PC) or nonproliferations or Senior-Looked syndrome. Polycyclic kidney disease can occur when there is defect in the cilia of the renal tube cells. Another genetic disorder called Barded-Bided syndrome (BBS) occurs hen the mutant gene products are in the components in the basal body and cilia. The symptoms of these diseases include physiological defects such as kidney failure, defective mucus clearance, neurosurgery deficits, and obesity. Defects in cilia or flagella in men sperm can lead to male infertility if the sperm cannot successfully move up the fallopian tube. Lack of functional cilia in female Fallopian tubes can cause octopi pregnancy, which is when the embryo implants outside of the uterine cavity. The cause of cilia defects is not really known other that genetic mutations. The cilia defects can get passes down from generation to generation. Symptoms for defects are not particularly known, but are circumstantial from defect to defect. Chlamydeous is a green plant that reproduces asexually and sexually. Chlamydeous is known for a number of different characteristics. First of all, Chlamydeous is a haploid organism. This means that it only has one allele, thus only showing one genotype for the cell. This characteristic makes it very easy during experiments to know why certain traits and description are a certain way and it makes it easy to test for variables. Chlamydeous are also known for having a generation time of two weeks (from mating of one generation to when this next generation can mate), meaning that it is easy to see results on a generation of Chlamydeous if it was test with a certain protein, as compared with the control. Chlamydeous has two interiorly inserted whiplash flagella that are similar to human cilia and contain the same flagella as humans. Chlamydeous also is not a new model organism, meaning that there are many different mutant strains that have unique characteristics from one another. Those unique characteristics sometime mimic environments inside the body and can be used to see how the body will react to the variable without actually affecting a patient. Victor Stool and his colleagues recognized that cilia play critical roles in animal physiology and development. They need to identify the gene products necessary for cilia assembly and function by running a proteomics analysis of human cilia. However, the proteomics analysis of human cilia is limited in its ability to detect low-abundance proteins. Comparative genomics was used to identify genes mound only in the genomes of organisms with cilia and flagella and was successful. This study was then implemented to find the low-abundance proteins of human cilia. Stool now found a use for the Chlamydeous because one, it has similar flagella or cilia to humans, two, most known components of cilia and flagella are strongly induced during flagella regeneration in Clampdowns and three, most of the identified human cilia disease genes have orthogonal in Chlamydeous that have been shown to be involved in flagella assembly. During process of flagella regeneration, it has been shown that many known legal components are transcriptional induced, with most transcripts reaching maximum accumulation between 30 to minis. Supplied with the Chlamydeous genome, Stool and his team set out to analyze the RNA transcriptional profile observed during flagella regeneration to identify flagella genes that have its RNA transcription activated in response to flagella regeneration. HASPS is the protein that our lab group took interest in. HASPS was up- regulated during flagella regeneration and chosen as a protein of interest. Hasps are called 70 kiloton heat shock proteins and are a family of heat hock proteins. Hasps are important for protein folding and essentially protects the cell from stress. HASPS-A or pithier, is an inhibitor of app-mediated optimism and app-dependent gene transcription such as cyclic G, app/waft, and MD expression. Our group wants to know if pithier acts as an inhibitor in the case of flagella regeneration in Chlamydeous. Based on the findings of last years Bio 111 lab group, they could not fully determine if fitting because on some cases flagella did fully regenerate, but on others it did not. Methods Chlamydeous Defoliation and Fixation Chlamydeous cells were defalcated using the pH shock method. Specifically acetic acid was added to the Chlamydeous growth medium in order to obtain a pH of 4. 5 resulting in the defoliation of the Chlamydeous cells. Defalcated cells were then immediately divided into one of three conditions. The two inhibitor conditions mixed 1 ml of cells with either a low (10 um) or high (100 um) dose of HASPS-A. In the third, control condition 1 ml of cells was mixed with 10 LU of ethanol. HASPS-A was dissolved in an ethanol vehicle; therefore, ethanol was added to the control condition in order to ensure that this ethanol id not become a confounding variable in the study. Immediately following the division of cells into the three conditions, 50 LU_ of each group was then fixed with equal parts Logos fixative. Logos fixative was used to kill, stain, and immobilizers the Chlamydeous cells to allow for measurement of the flagella regeneration. This first group of fixed cells represented time 0 of the study. This process was repeated every 15 minutes for a total of 90 minutes yielding 7 time points for each condition. As Chlamydeous is a photosynthetic organism the live samples were kept under deal light conditions in between data collection in order to ensure proper biological functioning. Non-Defalcated Chlamydeous One set of Chlamydeous was left untreated with acid in order to determine normal flagella length. These cells were also fixed with equal amounts Logos Fixative. However, non-defalcated Chlamydeous samples were only collected at one time point yielding a single baseline flagella length. Counting and Measuring Chlamydeous Flagella Lengths The Vicissitude program was used to photograph the first 20 Chlamydeous cells located on fixed slides from each time interval.