Expression of TDP43 in Development of ALS Model Zebrafish

Expression of TDP43 in Development of ALS Model Zebrafish Change in expression of TDP43 in various organs during development of ALS model zebrafish. Anuj Dhoj Raut Introduction Amyotrophic lateral sclerosis (ALS) is the devastating motor neuron disease that is characterized by progressive degeneration of both upper and lower motor neuron that control voluntary movement of body. The degeneration of the neurons seen in ALS result in muscle weakness, spasticity and atrophy of both cranial and spinal nerves muscle groups. Since there is often respiratory muscle involvement, aspiration pneumonia is the most common cause of death for the patients with ALS. At present, ALS is invariably fatal disease with no absolute cure and patients usually die within 3-5 years after the clinical onset of symptoms. The mean age of onset of ALS is between 55 and 65 years with slightly more prevalence in male (Male: Female ratio ~ 1.5:1) (1). Even though, incidence rate of ALS are different in different countries of the world, globally average annual incident rate is between 1.5 and 2.5 per 100,000 populations. There has been an increase in death rate of ALS and current internatio nal death rates for ALS have be close to 1 per 100,000 population per year(1). Currently, riluzole, an inhibitor of glutamate release, is the only disease modifying treatment available for the disease and can extends life only for couple of months (2,3). The etiology of ALS is currently unknown. However, approximately 10% of ALS patients have family history for ALS (Familial ALS;FALS) and remaining 90% of case occur sporadically (Sporadic ALS; SALS)(4). Although definitive evidence for environmental factor that cause ALS has remain mostly unknown, the evidence of genetic alternation that cause ALS has been increasing. Till date, only known cause of ALS is mutation in the gene. Mutations in more than 13 different types of genes have already been identified that can cause FALS. FALS is often a Mendelian inheritance with high penetrance, although most cases are autosomal dominant pattern of inheritance, autosomal recessive pedigrees have also been reported (5,6). Even though, FALS are cause due to genetic alternation, FALS are indistinguishable from SALS form histopathological perspective and both the types’ presents with similar sign and symptoms, thus suggesting common intra-cellular processes that lead to the disease symptoms. Among those 13 different types of gene mutation that causes FALS, mutation in Transactive response DNA binding Protein 43kDa (TDP-43) gene is seen in approximately 4% of FALS and 2% of SALS (7). Transactive response DNA binding protein 43kDa (TDP-43) is a DNA/RNA-binding protein encode by the TARDBP gene on chromosome 1. TDP-43 is an ubiquitously expressed nuclear protein capable of shutting between the nucleus and cytoplasm (8). TDP43 is present in almost all the tissue of a body and have different roles in different tissues (9). Although the precise cellular function of TDP-43 is unknown, TDP-43 has been implicated in regulating of gene transcription (9),alternative exon splicing (10) and mRNA stability (11). Under normal physiological conditions, TDP-43 resides predominantly in the nucleus where it involved in gene expression. But, in abnormal pathological conditions such as ALS, TDP-43 is mislocalized in the cytoplasm as inclusions body (12,13) . Analysis of TDP-43 in the brain and spinal cord of ALS patients reveled that TDP-43 is pathologically modified and redistribution to the cytoplasm, which is accompanied by loss of normal nuclear function and a toxic gain-of-function in the cytoplasm (14,15). The mislocalization of TDP-43 into cytoplasm is believed to be cause of neuron loss in ALS patients. Moreover, TDP-43 positive inclusions are also found either independent or partially colocalize with the other characteristic inclusion, such as tau, ÃŽ ±-synuclei, ÃŽ ²-amyloid and polyglutamines, which are found in other neurodegenerative disease such as Alzheimer’s disease, Pick disease and Parkinson’s disease. Interestingly, TDP-43 positive cytoplasmic inclusion are found in almost all ALS patient along with other neurodegenerative disease (16). Although evidence suggest that there is a definitive association between ALS and TDP-43, above observations make it confusing to whether TDP-43 pathology is causative or a secondary response in this disease. Studies done to unravel if TDP-43 is pathology or secondary response to ALS have come with conflicting result. Moreover, the present of TDP-43 in inclusion body of another neurodegenerative has been a mystery. The precise role of TDP-43 in ALS and other neurodegenerative disease is not well known and needs further evaluation. Study, in the mouse has shown that TDP-43 protein is essential for normal prenatal development. Homozygous loss of TDP-43 in mouse cause early embryo death. But, in heterozygous loss TDP-43 mouse, the TDP-43 protein levels were nearly normal suggesting an auto-regulatory mechanism controlling this protein levels(17,18). Moreover, research on mRNA expression levels of TDP-43 protein in various tissues has shown that TDP-43 plays different roles in different tissue(9). Furthermore, about 40 different mutant in TDP-43 have already been identified so far that is associated with ALS (10). But all this various types of mutations in TDP43 have only affected motor nerve of spinal cord and brain. At the same time, mutation and/or overexpression of TDP-43 has not cause any pathology alternation in other cells and tissue of the body or has been found to be associated with diseases of other organ system. A protein that is so vital for a development of organisms that it’s absent cause deat h, but when there is mutation in its gene has only abnormalities in nervous system and that abnormalities are evidence after mid-life is yet to be understood. Moreover, within the nervous system mutation in TDP-43 seems to affect only motor neuron and at the same time spares other neuron such as sensory, autonomic nervous system. And this preference to the motor neuron by mutant TDP-43 is even seen till the late stage of the disease. Physiological roles of TDP-43 and early cellular pathogenic effects caused by disease associated mutations in differentiated neurons is yet to be fully understand. Causative link between TDP-43 positive inclusion and ALS can be well established, if nuclear to cytoplasmic expression of mutant TDP-43 could be study in vivo and in real time. And at the same time, will also be able to understand if TDP-43 pathology is causative or a secondary response to ALS and other neurodegenerative disease. Transgenic rodent models of ALS have been extremely valuable in providing some insight into biological mechanisms underlying ALS. But, due to difficulty in conducting in vivo real time study with rodent, change in intra cellular expression of TDP-43 has not being well understand. The zebrafish has recently emerged as powerful genetic model system for studying ALS. External development and transparency make it great tool to study the development stages of almost all the organ. External development of its eggs allows easy observation and manipulation of early development process. And, transparency makes is a powerful tool to observe the change at cellular level by using fluorescent reporters. With the help of fluorescent reporter, specific cell type and protein expression within those cells can be easily identify and study in vivo and in real time in zebrafish. In addition, zebrafish is a vertebrate and their nervous system is highly conserved with higher vertebrates including humans a nd many pertinent feature of the nervous system start to develop within 1 day of development. Moreover, genetic manipulations are comparatively easy in zebrafish. Therefore, zebrafish is a great model system to study the association of TDP-43 and ALS. In this study, I am trying to understand the change in expression of mutant and overexpressed TDP-43 protein in different tissue of zebrafish. At the same time also will be evaluating the change in expressions of TDP-43 as the zebrafish grow from embryo to adult. I will then compare the change in level of TDP-43 from asymptomatic stage of ALS zebrafish to that of symptomatic stage of ALS zebrafish. In order to conduct this experiment, transgenic zebrafish with human mutant TDP-43 will be created by genetic engineering. Human mutant TDP-43 will be fused with green florescent protein (GFP) before creating transgenic zebrafish. By combining human mutant TDP-43 with GFP will allow easy visualization of TDP-43 protein in zebrafish. Then, image of the fluorescent labeled TDP-43 at different stage of development of zebrafish period will be capture with fluorescent microscope. References 1.Logroscino, G., Traynor, B., Hardiman, O., Couratier, P., Mitchell, J., Swingler, R., and Beghi, E. (2008) Descriptive epidemiology of amyotrophic lateral sclerosis: new evidence and unsolved issues. Journal of Neurology, Neurosurgery Psychiatry 79, 6-11 2.Bensimon, G., Lacomblez, L., and Meininger, V. (1994) A controlled trial of riluzole in amyotrophic lateral sclerosis. ALS/Riluzole Study Group. The New England journal of medicine 330, 585-591 3.Miller, R., Mitchell, J., Lyon, M., and Moore, D. (2007) Riluzole for amyotrophic lateral sclerosis (ALS)/motor neuron disease (MND). Cochrane Database Syst Rev 1 4.Pasinelli, P., and Brown, R. H. (2006) Molecular biology of amyotrophic lateral sclerosis: insights from genetics. Nature Reviews Neuroscience 7, 710-723 5.Mulder, D. W., Kurland, L. T., Offord, K. P., and Beard, C. M. (1986) Familial adult motor neuron disease: amyotrophic lateral sclerosis. Neurology 36, 511-517 6.Gros-Louis, F., Gaspar, C., and Rouleau, G. A. (2006) Genetics of familial and sporadic amyotrophic lateral sclerosis. Biochimica et biophysica acta 1762, 956-972 7.Corrado, L., Ratti, A., Gellera, C., Buratti, E., Castellotti, B., Carlomagno, Y., Ticozzi, N., Mazzini, L., Testa, L., and Taroni, F. (2009) High frequency of TARDBP gene mutations in Italian patients with amyotrophic lateral sclerosis. Human mutation 30, 688-694 8.Winton, M. J., Igaz, L. M., Wong, M. M., Kwong, L. K., Trojanowski, J. Q., and Lee, V. M.-Y. (2008) Disturbance of nuclear and cytoplasmic TAR DNA-binding protein (TDP-43) induces disease-like redistribution, sequestration, and aggregate formation. Journal of Biological Chemistry 283, 13302-13309 9.Ou, S., Wu, F., Harrich, D., Garcà ­a-Martà ­nez, L. F., and Gaynor, R. B. (1995) Cloning and characterization of a novel cellular protein, TDP-43, that binds to human immunodeficiency virus type 1 TAR DNA sequence motifs. Journal of virology 69, 3584-3596 10.Lagier-Tourenne, C., Polymenidou, M., and Cleveland, D. W. (2010) TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration. Human molecular genetics 19, R46-R64 11.Strong, M. J., Volkening, K., Hammond, R., Yang, W., Strong, W., Leystra-Lantz, C., and Shoesmith, C. (2007) TDP43 is a human low molecular weight neurofilament ( h NFL) mRNA-binding protein. Molecular and Cellular Neuroscience 35, 320-327 12.Arai, T., Hasegawa, M., Akiyama, H., Ikeda, K., Nonaka, T., Mori, H., Mann, D., Tsuchiya, K., Yoshida, M., and Hashizume, Y. (2006) TDP-43 is a component of ubiquitin-positive tau-negative inclusions in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Biochemical and biophysical research communications 351, 602-611 13.Mackenzie, I. R. (2007) The neuropathology of FTD associated with ALS. Alzheimer Disease Associated Disorders 21, S44-S49 14.Kabashi, E., Lin, L., Tradewell, M. L., Dion, P. A., Bercier, V., Bourgouin, P., Rochefort, D., Hadj, S. B., Durham, H. D., and Velde, C. V. (2010) Gain and loss of function of ALS-related mutations of TARDBP (TDP-43) cause motor deficits in vivo. Human molecular genetics 19, 671-683 15.Neumann, M. (2009) Molecular neuropathology of TDP-43 proteinopathies. International journal of molecular sciences 10, 232-246 16.Da Cruz, S., and Cleveland, D. W. (2011) Understanding the role of TDP-43 and FUS/TLS in ALS and beyond. Current opinion in neurobiology 21, 904-919 17.Kraemer, B. C., Schuck, T., Wheeler, J. M., Robinson, L. C., Trojanowski, J. Q., Lee, V. M., and Schellenberg, G. D. (2010) Loss of murine TDP-43 disrupts motor function and plays an essential role in embryogenesis. Acta neuropathologica 119, 409-419 18.Sephton, C. F., Good, S. K., Atkin, S., Dewey, C. M., Mayer, P., Herz, J., and Yu, G. (2010) TDP-43 is a developmentally regulated protein essential for early embryonic development. Journal of Biological Chemistry 285, 6826-6834

Spot The Difference Attitudes towards people onwelfare benefit in the 19th and 21st centuries

To be clear on the people who receive welfare benefits, it is necessary to divide everyone into classes. Even if people refuse to admit they are in a ‘class' there are clear characteristics of what class they belong to. This division that is still active now, was even more operational in the 19th century. It was a way of life that the higher you were in the social class and hierarchy, then the more successful and prosperous you were to become in life. It was also some times a case of where you lived. It was Charles Booth that marked on maps of London where each social class lived. Places like Mile End Road and Orsman Road contained the ‘vicious poor', the people at the bottom of the hierarchy. They were labelled as; ‘The lowest class which consists of some occasional labourers, street sellers, loafers, criminals and semi-criminals. Their life is the life of savages, with vicissitudes of extreme hardship and their only luxury is drink.' Although this may seem that Booth is being ‘snobbish', it was noted that Booth had sympathy for the poor. He worked with the lower class, and reported that it wasn't always necessary to have money to be happy. He described that although the poor were more likely to die of disease and less likely to survive, he thought that they seemed to be happier, without nurses and servants etc. That the rich are more likely to suffer from being spoiled than from harshness, ‘that the simple natural lives of working-class people tend to their own and their children's happiness more than the artificial complicated existence of the risk.' Now in the 21st century, these locational divides are still in place, although the locations of the ‘vicious' poor have changed. While it is more noticeable in the 19th century maps, the upper class and lower class virtually side by side, it is possible that people were more willing to tolerate each other, nowadays there are more clear divisions. This change could be due to people's tolerance and attitude towards different classes. However, it seems more apparent that there are different attitudes towards social classes. Usually, people aren't willing to live near people of a lower class, places like council estates accommodate people of the same status, but they are prepared to pay taxes towards their welfare and benefits. During the 1800's the conditions of the workhouses, and the ‘relief' from the government or parishes was of little help. People still struggled to make ends meet. The rule was that no one got above the lowest workers wage, which was 12s to 15s a week. It was said that for a comfortable life, a worker needed a wage of 30s a week, concluding that few people had a comfortable life. In 1885, it was reported that 25% of the population lived in poverty, however, after Charles Booth investigated, and wrote Labour and Life of the People, he found that actually 35% of the population were in poverty. In simple terms, it was harder to receive help in the 19th century. Today there are fourteen types of benefits, including: benefits in kind for employees, child maintenance, council tax benefit, disability and carers benefits, housing benefit, incapacity benefit, income related benefits, invalid care allowance, mothers, widows and families benefits, retirement allowance, statutory sick pay, unemployment benefit, unfit for work benefits and war pensions and industrial injuries. While in the 19th century, it was an innovation to have the poor law, today there are 2.7 million people claiming incapacity benefit, and the government are trying to reduce this number. 1.610% of the population are on incapacity benefit; this is an immense difference from what it was in the 1800's. Although there are so many types of benefits, the government has become more active in getting people into work. The attitude of the government is that they need as many people in work as possible; they have introduced schemes such as EMA, something that would never have been thought of in the 19th century. People's attitude was that the government were doing something about the increasing poverty, at the expense of the economy. But why is the government making changes to benefits? The bottom line is that society has a responsibility to care for those unable to work. The government have introduced new schemes to prevent people from abusing benefits, the same thing that the government did in the 1800's when welfare was introduced. The changes proposed are likely to separate the seriously disabled or those suffering from terminal conditions such as cancer, who are unlikely ever to return to work, from those claiming to be incapacitated by a â€Å"bad back† or depression. People's attitudes have changed enormously from the 1800's to the present day. In the poor law days, going into the workhouse was shameful; people did as much as they could to prevent this, it was the lowest they could go. The workhouse conditions were terrible, starvation was often a common factor, families were separated and people's dignity and rights weren't an issue. People's attitudes today have changed a great deal over the years not because welfare benefits have changed, but because people's basic human rights have become more of a factor. It is expected that people who cant feed and look after themselves or their family, can receive help from the government. This way of thinking has developed partly from the original poor law. People started thinking that they needed to help others, even if it meant paying in taxes. Today, although people aren't ‘proud' of receiving benefit, it has become more acceptable; it is possible for people to stay on benefits their whole lives however, it costs the government à ¯Ã‚ ¿Ã‚ ½12 billion a year to fund benefits. This extra spending has been criticised by certain groups. There a lot of differences between 19th and 21st century welfare benefit. People's attitudes today mean that it is common for people to receive benefits. They know that they can fill in a form and receive at least à ¯Ã‚ ¿Ã‚ ½55 a week, not including child allowance. Today people can live just as well as people who work, which has caused some protest. While it seems that in the 19th century, welfare was a last resort, they didn't want to receive help, partly because the standard of help sometimes wasn't better than being left to starve. From old maps of London, it can been seen that people used to live close to others of different classes, while today, it is more likely that people move to places that are within the same income bracket. This displays another way that attitudes have changed, that people aren't willing to live near people who cannot support themselves, or they live near people of similar means. However, some similarities can be found, although it can be assumed that today attitudes have become more relaxed, today's government tries to remain vigilant as it was years ago. The government are aware of people mistreating the benefit, and so have chosen the attitude to fight those who misuse it. This could include imprisonment and fines etc. People had more of a superior attitude towards people on benefits in the 1800's, it was assumed by some that these poor were too lazy to work and the same can be said for today. But the underlying principle still remains, in the 19th century and 21st, welfare benefits are aimed to help people, and although people may have different feelings about those dependant on welfare, the benefits will still remain in place.

Dehumanization in Night Essay

Night is a heart pulling memoir of its young Jewish author, Ellie Weasel, and his experiences in the Holocaust. The book begins with him living in the town of Sighet. He had a very sheltered life, with no accounts of negativity in the world. He and his family were also raised heavily on Jewish beliefs. One day a man by the name of Moshe the beadle comes to warn the people of the dangers of the Nazis. Unfortunately the people did not heed this and Sighet was invaded by Nazis. Weasel and his family are taken and separated. He only had his father now and they braved much torture and mal treatment by the kapos in the camps. At the end of it all only weasel himself made it out alive, though a brutal scar was marked upon his soul. He’d lost his family and his faith at those camps. But through all his sorrow and loss he wanted to share his accounts in this dark volume of his life, so that people understand what the Jews went through all those years ago. This led him to write Night, where in which Weasel points out the inhumanity towards other humans during the holocaust as one of the themes of his chilling story. One of the major factors that contribute to this theme is actually one of the first cruel things he encountered was the Nazis. At first on the other had he didn’t see them for the monstrous people that they were. In the book Eliezer, Weasels character, even recalls, â€Å"Our first impressions of the Germans were most reassuring†¦. Their attitude toward their hosts was distant, but polite.† But this is just one of the many aspects of the holocaust that was tremendously misunderstood. But even more so unthinkable was the cold-blooded butchery of millions of innocent people. As the memoir progresses you will see how Weasel puts a spotlight of the actions of the Nazis by first seeing them as humans beings but then later on reveals the evil deeds that they commit upon innocent Jews. Night also exhibits how inhumanity can spread to others who have been shown inhumanity. This is shown when the Jews start to turn on each other, instead of braving their harsh treatment together. It is even said by a Kapo: â€Å"Here, every man has to fight for himself and not think of anyone else. . . . Here, there are no fathers, no brothers, no friends. Everyone lives and dies for himself alone.† Because the Kapo are also just prisoners that are in control of the other prisoners, this is a very significant message. They were happy to help the Nazis in their plans for genocide. This led them to act really ruthlessly towards those under their command. In the fifth section of the book Eliezer mentions them as being, â€Å"functionaries of death.† The perspective of the Kapos show how those effected by the Holocaust can use inhumanity to infect other people like it as a virus.

Social Networking The Best Guinea Pig - 2083 Words

The purpose of social networking was originally meant to open our eyes to the world, not blind us from reality. As the first generation to grow up with social networking, Generation Y is the best guinea pig to observe how humans begin to interact with one another while in constant global communication. Social networking has enabled the world to unify, spread ideas, communicate, and spread news quicker than ever before. Humans are now able to stay in touch while in completely different places. There is no more need to send letters or wait until the next day to read the newspaper because social media offers an instantaneous ability to inform the individual and the public. Additionally, people are now able to meet new â€Å"friends† through†¦show more content†¦This is where social media came in use. In order to spread the word for help people began tweeting out to map coordinates to inform the rescue teams where a person in need was. Also, the survivors were able to uti lize Facebook in order to link up with loved ones who were separated from the natural disaster (Howerton). Due to social networking many more lives were able to be saved that may have been lost without it. It provided a fast and easy ways for rescue teams to locate people in need. Social media is important because it always a quick response to emergencies. Also, it provides an emergency prompter to inform the general public of a possible incoming disaster. Next, scientists have been able to use social networking in order to share their experimental data. In the effort to find all the mutations associated with cancer many scientists felt it better to work together in order to share data. As a result, when â€Å"sequence data would come off the machine†¦ it was immediately posted on internet accessible data sets that could be accessed by all the investigators in the project as well as other scientists interested in looking at the data† (Sawyers). Without the ability to ins tantly share information scientists would not be able to have the most up to date information at all times. Investigators are now able to share their information, successes, and failures with others in order to keep modifying science. Medicine is at a great advantage because now doctors can easily