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An Experiment to Investigate Effect of Ageing on Role of EC Cells and 5-th in Gastrointestinal Motility - Coursework Example

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This coursework "An Experiment to Investigate Effect of Ageing on Role of EC Cells and 5 Gastrointestinal Motility" describes a mouse of various ages to examine the role of EC cells and 5-HT in motility as well as their characterization and effect of age on these cells…
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AN EXPERIMENT TO INVESTIGATE EFFECT OF AGEING ON ROLE OF EC CELLS AND 5-HT IN GASTROINTESTINAL MOTILITY Name: Course Professor’s name University name City, State Abstract The contribution of the effect of age on the function of colon leading to disorder and constipation in human is incompletely understood. Accordingly numerous recent studies have shown 5-hydroxytryptamine (5HT) is to be essential for initiating reflexes resulting from mucosal stimulation and also for normal propagation of colonic migrating motor complexes (CMMCs) together with propulsion of faecal pellets in isolated colon. Evident changes with age occurrence spontaneously in human such as colonic dysmotility and constipation have raised the question about the effect of ageing on enterochromaffin (EC) cells, which may suggest age related issue in function of these cells. This study is designed and uses a mouse of various ages to examine the role of EC cells and 5-HT in motility as well as their characterization and effect of age on these cells. This study comprise; 1) isolation of EC cell by immunocytochemistry and serotonin content which involves a combination of enzymatic digestion, staining, short term cell culture and density gradient centrifugation, 2) Evoke inherent mucosal reflexes tested by brush stimulation or circumferential stretch, 3) recording from electrical field stimulation of segments of intact colon with and without mucosa. Electrochemical technique is used in this study for recording the 5-HT release selectively and quantitatively and to obtain the measurement of steady state 5-HT level from the isolated cell as well as 5-HT produced by mechanical stimulation. Furthermore this study will clarify the role performed by change in 5-HT release properties with age by informing of the likely changes in humans and impact on human health in relations of understanding the physiological and pathological changes and influences therapeutic approaches to some of the conditions and facilitating in an increase in more study within these area of research. Introduction The world is on the verge of demographic milestone. According to (Richard , 2012), the number of people aged 65 and above has outnumbered children below 5 years. This has been attributed to remarkable rise in life expectance, falling fertility levels and advanced medical care. With fewer people joining the population and older people living longer, older population are continuously making the larger share of the demographic data. Despite the projected uncertainties connected to all population estimates, the global pattern of demographic change is expected to change a lot in the net half century, with ageing being the key feature of population dynamism., and the pressure on the public expenditure being intense in those areas of economy which general deal with the aged population, such as health and pension schemes. As expected ageing leads to substantial rise in age- related expenditure by the government. Equally startling is the potential difficulty by governments in keeping up with the ballooning pension schemes and health care budgetary estimates (Forssmann, 1976). Another key impact of fast ageing population is the potential downturn in living standards as a result of capital accumulation. This will be as a result of increased rates of labor productivity growth emanating from the increasing capital stock (Hagemann, & Nicoletti , 1989). An increase in population will also strain the existing medical facilities. The effect will be more pronounced in more so in developing countries who are also experiencing an overstretched public health facilities (Haque, & Monteil, 1989). According to (Mangoni & Jackson, 2004), advancement in age is characterized by the impairment of the majority of the bodies’ regulatory functions that provide the functional integration of the cells and organs. Technically, ageing is the effect of these underlying alterations and changes and not the cause. The most consistent definition of ageing is the time- related decline of performance of the functional units in body (Bertrand, 2003). With rise in age comes the likelihood of disability. This is due to the fact that is people live much longer, and do not suffer fatal illness, they are likely to get chronic illness instead. (Neena & Heather , 2014), asserts that the interaction between age advancement and increasing physical disability thus leading to a negative image of ageing. Literature review The occurrence of certain gastrointestinal disorders, such as constipation and dysphagia, increases coincidentally with age (Jin , 1999). Changes in the neuromuscular and gastrointestinal function, as a result of aging have been established in models of ages in both humans and ageing. (Hall, 2002) The result of age on the human colon anatomy and function is not well understood. The prevalence of stomach disorders in humans such as diarrhea and constipation is more prevalent with age. ( Grider, 2003) Performance of organs generally deteriorates or changes with age. As a result majority of these organs depict structural changes as well as diseases and disorders as age progresses. Typical to these organs, disorders and diseases in adult gastrointestinal tract (GI) are more prevalent as age increases. However, generally, the gastrointestinal tract ages well thus being predisposed to certain diseases and disorders. (Cheryl , et al., 2010), a point out colonic transit generally slows down with age, however most studies have been conducted in animals rather than in humans., and data from humans are ambiguous , showing general slowing down of colon transit and in some cases no change ( Bertrand & Kunze, 2000). The high prevalence in constipation with age can be as a result of several reasons, among them being reduced mobility, increased usage of medication, alterations of food and fluid intake as well as enteric nerves changes. Loss of enteric is experienced with age. A number of research studies have concluded that there occurs a loss in neurons with age in guinea pigs, rats, mice and humans. (Hall, 2002), concludes that for instance in the rat, the loss in enteric neutrons with ages is not typical to neurons subtypes however it appears to be limited to those neutrons that express choline acetyltransfarese (ChAT) The selective enteric neutrons loss of ChAT is coupled with a sparing neutrons expressing neuronal nitric oxide (nNOS). (Cheryl , et al., 2010), assert that similar results were shown by different strains rat, proving rather generalized findings in the rat. The selective loss of ChAT- positive was attributed to a progressive increase in inhibitory neurons as the age of the animal increases, resulting in the alteration of the colonic function. (Cheryl , et al., 2010), also believes that it is still unclear if the loss also occurs in human colon with age. According to (Cheryl , et al., 2010), there is one study conducted to investigate the impact of aging on neuronal cell number present with the human colon myenteric ganglia. In the preliminary study conducted on the necropsy specimens, the quantities of the myenteric neuronal cells were counted with the help of Giemsa stain in two age groups of 65 years and35 years old. It was concluded that there were changes in ChAT and nNOS positive neurons in both volume and plexuses of the human colon across different ages. Functioning of the GI tract (Neena & Heather , 2014), asserts that since the digestive system has a lot of in-built reserve, it has the capability of protecting itself, thus aging will have little effect on its functionality of compared to other organ systems in the body. Nevertheless, aging can be a factor attributed to certain other disorders within the gastrointestinal tract. Above all, older adults potentially likely to develop digestive tract disorders and diverticulous as a side effect of certain chemicals contained within the drugs. With age the esophagus, tension and contraction experienced in the upper esophageal sphincter tends to decrease, however (Hagemann, & Nicoletti , 1989), suggests that such changes are not known to impair with food movement along the gut. However, aged adults are most likely to experience certain diseases which interfere with the normal esophageal contractions. In the stomach section of GI, age brings in a condition where the stomach lining capacity undergoes perennial damaging. This in turn increases the risk of the person developing peptic ulcer diseases, especially those who non-steroidal ant inflammatory drugs such as aspirin. Also due to decreased elasticity, the stomach of an aged person lacks the capacity to accommodate more food; also the rate at which food is maintained from the stomach into the small intestines greatly decreases. This GI changes however, do not show noticeable symptoms. According to (Richard , 2012), although aging does not interfere with the effect on secretion of the acids and peptides, however the conditions that leads to the in reductions have been attributed to other reasons such as atrophic gastritis. In the small intestines, there are minor effects felt as a result of aging. Therefore movements of intestinal through the small intestines and absorption of most nutrients remains unchanged over the life. The lactose level decrease leading to more lactose intolerance. There is also a probability of growth of certain bacteria, which leads to bloating, pain and general weight loss. High bacterial concentrations may also lead to reduces absorption of certain vital nutrients such as calcium, folic acid and iron. According to (Lovat, 1996), there are no age related alterations in the anatomy of the small intestines and height and count of enterocyte and intraepithelial lymphocyte remain unchained. According to (Lovat, 1996), a study carried out on rodent revealed that an increase in cell proliferation was recorded in response to injury. If the same can be replicated in humans, then it’s possible to conclude that there can be a resultant increased susceptibility to gastrointestinal cancers within the aged populations. The contribution of the effect of age on the function of colon leading to disorder and constipation in human is incompletely understood. Accordingly numerous recent studies have shown 5-hydroxytryptamine (5HT) is to be essential for initiating reflexes resulting from mucosal stimulation and also for normal propagation of colonic migrating motor complexes (CMMCs) together with propulsion of faecal pellets in isolated colon. Evident changes with age occurrence spontaneously in human such as colonic dysmotility and constipation have raised the question about the effect of ageing on enterochromaffin (EC) cells, which may suggest age related issue in function of these cells. Within the aged, healthy small intestines maintains their absorptive nature of colon and duodenum of starch with no alterations in the duodenal membrane enzyme activity of glucose movement. However, the absorption of vitamin B12, remains unchanged within the elderly health humans. The frequent malabsorption within the elderly does not seem to be specifically concluded to be age related. Constipation is common in the elderly. This is attributed to slight slowing down of the GI contents movement and a decrease in rectum rhythmic contractions. Oracaceal transit time does not generally change as a result of age. Although, with age the GI of old becomes sensitive to thyroid hormone status, also colonic transit generally slows down with ageing, although the effects are highly variable. There are no much changes in the large intestine as a result of age. However due the age advancement, the rectum somehow enlarges. Other than tumor cells and stem cells, ageing is virtually a universal process, whereby through functional decline, there occurs cell death, and ultimately death of the organism, (Hagemann, & Nicoletti , 1989), asserts that molecular changes as a result of age within the human colon have been adequately determined; in addition, the existing research studies, results and conclusions after tests in animals and humans are in most cases contradictory and ambiguous. A number of changes in both colonic and rectal functions as a result of ageing have been identified. From the conclusions drawn (Gabella , 2001), the total quantity of colonic myenteric neurons reduces in age within children and rats, especially during the first 4 years. According to (Graff, et al., 2001), noted a rise in surface area of myenteric ganglia with age. However, (Cheryl , et al., 2010), found ganglia proportion with structural abnormalities such as cavities increased with age. Another noted correlation was the collagen content and age within myenteric ganglia. (McDougal, et al., 1984), concludes these alterations within the colonic innervation may be the cause of effects on colonic motility. According to (Graff, et al., 2001), colonic epithelium is renewed in every 4 to 5 days in humans. This epithelial regeneration is regulated is permitted by regulated balance of epithelial apoptosis and proliferation. Any nonconformity in epithelial cell kinetics may lead to loss to functional and structural integrity. The noted imbalance colonic epithelial regeneration may result to either development carcinoma or ulcer of the colonic mucosa. (Hall, 2002), concludes that the impact of ageing on the regeneration of colonic epithelial is not well understood. For instance, in colon of the rat, the proliferation of crypt epithelial were found active in week 3 of their life, which was according to (McDougal, et al., 1984), suggests it was thought to be connected with the developmental of the gastrointestinal tract. Conclusions according to ( Loening-Baucke & Anuras , 1984), are on the other hand contradictory; with the quantity of proliferative epithelial walls greater, while the apoptosis lower in aged rates. This phenomenon can be attributed to explanation that the survival of defective genetic cells; thereby increasing the frequencies of colorectal cancer amongst the elderly population. According to (Ferenc & Katalin , 2011) pathologic and physiologic as a result of age in the human colon impact both regenerative capacities of the epithelium as well as inflammatory resulting in the mucosal injury. In contrast, the altered function and migration of the regenerative stems cell, and the methylation of the mucosal healing due to age, coupled with the alterations of growth factor may be involved in deferred in mucosal regeneration. However, according to the (Lee , et al., 2000), asserts, the entire connections of the above interactions with the age process are yet to be fully understood. The understanding and custom personalized modifications of these interactions are of clinically vital in respect to modern therapeutic strategies and prevention of diseases. The rise in cell proliferation due to age can be attributed to enhanced transition from GI phase to S phase, and any inconsistent may be as a result of different locations and animals sampled. Some of the inflammatory bowel diseases such as Crohn’s disease, ucerative colitis (UC), are some of the reasons for epithelial destruction within the colon. Effects of ageing on colonic healing The luminal border of the colon is lined with an epithelial layer which serves a number of functions, such as electrolyte and water absorption, as well as a barrier for harmful luminal pathogen (Forsberg, & Miller,, 1983). As a result of the high turnover of detaching epithelial cells, it is crucial for their replacement from the local stem pool of cells. It is well understood that circulating ste cells are play a vital role in hematopoietic homeostasis, and alos the regeneration of the solid organ cells. There is however, evidence for and against the ageing of stem cells, however most publications are not in agreement with the quantity and quality alterations of the stems cells during the ageing period of an organism (Chen, et al., 1997). The impact of ageing on colonic epithelial regeneration and crypt- base stem cell function has come to the frontline of recent discovery. For instance, in the apoptotic cells were discovered around the stem cells and this frequency did not alter with age. (Hagemann, & Nicoletti , 1989) , suggests that there are a number of signs of ageing within the gastrointestinal tract, such as colon. Far from microscopic and macroscopic alterations, some can be detected at gene expression, genetic as well as at epigenetic level. The interaction between colonic mucosal regeneration and ageing has been concluded by a number of studies, however such results only provide as short insight into the pathologic and physiologic mucosal healing within the ageing context. An understanding of the impact of the age- related mechanisms may assist in the rising new therapeutic approaches for chronic colonic diseases accompanied with mucosal destruction of enterocytes found in the mice; 2-4 days (Eamonn , et al., 2012). ECL cells are composed of over 30% of the neuroendocrine cell population of the gastric mucosa and are crucial in the peripheral regulation of the secretion of the gastrin acid (Mazzuoli . & Schemann , 2012). In responding to PACAP and gastrin, the ECL cells produce histamine in a calcium dependent design, thereby stimulating the secretion of acid from the parietal cells. The transport of the histamine through the vesicular monoamine transporter-2 affects the storage of the histamine within the secretory vesicles (Dante, et al., 2009). This hypergastrinemia lead to creation and development of the ECL cells tumour. Past research have concluded the presence of certain endocytotic and exocytic proteins within the ECL cells in addition to other neuroendocrine cells, thereby being a clear indication of a marker genes in pointing the characteristics traits during the microarray techniques. (Schäfermayer, et al., 2004), assert that such studies have been the benchmark for determining the dynamics during the inflammatory or malignant transformations within in the gut (Powell , et al., 2003). Sampling of macromolecular by the colon performs a vital role in both pathophysiological as well as normal physiology. Alterations in the Lower Gastrointestinal Tract as a result of Ageing (Lindsay & Miaskowski, 2008), asserts, that anatomic changes related to age within the lower GI tract may be the reasons for the delayed movement of pace for stool and associated water content. These changes include, decreased blood flow, changes in the intestinal wall atrophy, and alterations in the intrinsic neurons (Vanden, 2008). Other functions of GI such as absorptions and secretions generally remain unchanged. According to conclusions drawn by (Lindsay & Miaskowski, 2008), transition time in the gut and colonic mobility remained generally similar in healthy older adults when compared to children. In contrast, (Lee , et al., 2000), asserts that elderly people suffering from chronic illness, and who reported constipation, had a 4 to 9 days prolonged GI tract transit time, with more delayed experienced in the lower gut and rectum. (Hall, 2002), believes that colonic motility is influenced by factors related to ageing, such as immobility, medication and diseases. ( Bassotti & Villanacci , 2006), suggests that Age- related neurodegenerative alterations in the enteric nervous system (ENS), may be the reason for the functional changes realized with the advancement of age. For instance in in colons of humans aged more than 65 years, (Hall, 2002), noted a 37 percent decline of the enteric neurons as compared to younger humans. In the experiment, (Hall, 2002) used a laminar preparations of the muscular external to determine the number of nerve cells present in two groups of people ages 20, 35 and 65 years. On top of that, the elastic and collagen systems were more numerous within the ganglia of participants aged more than 65 years (Martel, & Monteiro, , 2003). From the conclusions, (Hall, 2002), noted the loss in neuron density as a result age is supplemented, by the apparent boost in the fibrous matter of myenteric ganglia. This above research findings suggests that changes in the neurodegenerative may contribute to the disturbed colonic motility experienced by the aged population. According to (Lindsay & Miaskowski, 2008), aged people have experience reduction in IAS pressures in both anal and pelvic muscle strength. This change in muscle strength eventually alters the anal function and rectal sensitivity. The problem is more pronounced in women, after menopause since they experience a more decline in squeeze pressures due to injuries sustained during child birth. The changes ultimately put them at a risk of potential constipation. Constipation so generally is not a straightforward condition. This is further compound by the fact that there are sparse research finding from recent research on the mechanisms and effects of constipation on the elderly. For proper treatment to be realized, it is best that mechanisms of constipations get thoroughly researched (Scherl, & Frissora, , 2003). Pelvic floor dysfunction and dysmotility are only pointed out mechanisms associated with constipation, disruption in colonic motility. Further research studies are therefore necessary in order to fully understand the physiologic, anatomic and lifestyle reasons that impact theses mechanisms. Additionally, longitudinal data on GI motility is requited in the determination of the impacts of ageing on the colon and lower gastro intestine (Chen, et al., 1997). (Lindsay & Miaskowski, 2008), suggests that the changes in the GI function, appear to be relatively unrefined based on the limited quantity of research findings collected and the extent of conflicting conclusions drawn. Additional research is therefore necessary to fully understand the effects of age on GI gut function, as well as the timings and impact of these changes. A deeper understanding derived from critical analysis of the changes in colonic wall in the older people could offer a new direction for both assessment and management of constipation and other bowel disorders within this vulnerable population (Modlin,, 2006). Aims and objectives The overall objective of this research is to determine the effect of ageing on role of EC cells and 5-HT in Gastrointestinal motility as well as their characterization and effect of age on these cells. Specific objectives i. Isolation of EC cell by immunocytochemistry and serotonin content which involves a combination of enzymatic digestion, staining, short term cell culture and density gradient centrifugation, ii. Evoke inherent mucosal reflexes tested by brush stimulation or circumferential stretch, iii. Recording from electrical field stimulation of segments of intact colon with and without mucosa iv. To give a summary of the status of the current researched knowledge concerning neorodegeneration of the enteric nervous system (ENS) in ageing and its functional implications v. Suggest areas where significant research advances are likely to yield vital knowledge in future The main hypotheses for this experiment a: H1- there is correlation between age and gastrointestinal disorders Ho- there is no correlation between age and gastrointestinal disorders 2.0 Methods and Materials Inherent mucosal reflexes tested by brush stimulation or circumferential stretch Experimental Protocol The isolated colon was removed from petri dish and fixed in organ bath by pining the entire colon as flat sheet preparation, to record motor activity of circular muscle layer. Protocol for Mucosal Stimulation The tissue was left to rest in water bath for 40min and subsequently challenged by applying 1-5 strokes to the mucosa with fine artistic paintbrushes, every 10 minutes. In this study each strokes have been repeated up to three times to ensure accuracy and precision. Drug and Solutions The tissue was challenged in present of antagonists, steps (…..) was repeated for each of the antagonist listed below. Antagonist added to the Krebs solution were (1μm scopolamine), (1μm GR159897) and (1μm Ondansetron). Reproducibility tests All of the experiments in this research were repeated five times with identical protocol for various age groups mentioned in this study to obtained reproducibility test. Analysis of data and statistical methods All of the data in this study were measured using MLT 0202 force transducers and recorded on a Power Lab/7sp data recording system with Chart v5.5.6 software Electrical field stimulation of distal colon with and without mucosa In this research male C57BL/6J mice obtained at 8 week of age from Harlan UK. The mice were kept in separate ventilation cage in groups of 3-4 under barrier-reared environments until required. Laboratory mice were maintained at 20 °C, 50% humidity, and were fed on maintained diet according to the guideline with free access to irradiated water. Additionally mice were retained on 12-hour light and dark cycle and studied at age of 3, 18 and 24 months. Electrical field Stimulation To determine tissues viability electrical field stimulation was performed by the Grass SD stimulator at constant maximal voltage of 80V with pulse frequency of (0.1,0.3.1,3,10 and 30Hz) for 30 second. The distal colons were allowed to recover for 4.5 min between each stages of frequency stimulation. Serotonin Dose Response The tissues were also been challenged for serotonin dose response by addition of the 250μl of serotonin dose concentration (mol/L) of (1x10-5, 3x10-5, 1x10-4, 3x10-4, 1x10-3, 3x10-3) into each chamber. In this experiment the serotonin dose response were recorded for both tissues for duration of one minute. Subsequently after each serotonin dose response the tissues where thoroughly rinsed with Krebs solution and left to recover for nine minutes. Analysis of data and statistical methods All of the data in this study were measured using MLT 0202 force transducers and recorded on a Power Lab/7sp data recording system with Chart v5.5.6 software. The data present as mean ± SEM and the statistical comparison of date were performed using two-way ANOVA with Bonferroni post-hoc test and student’s t-test. Results Inherent mucosal reflexes tested by brush stimulation or circumferential stretch The results observed were summarised as shown in the figure below The purpose of conducting this test was to determine whether a pellet controls the instigation of CMMC by activating local reflexes.. From the results in figure above the response was higher for control mucosa as compared with the presence of Ondansetron. Response as a result of control mucosa was also same for all the age groups i.e., 3, 18 and 24 months, all recording about 6g/s. This shows that CMMCs appear to begin orally to the pellet and ultimately propagate anally. Interaction accounts for 5.36% of the total variance. F = 2.85. DFn=8 DFd=60 The P value = 0.0095 If there is no interaction overall, there is a 0.95% chance of randomly observing so much interaction in an experiment of this size. The interaction is considered very significant. Since the interaction is statistically significant, the P values that follow for the row and column effects are difficult to interpret. AGE accounts for 13.01% of the total variance (after adjusting for matching). F = 2.61. DFn=2 DFd=15 The P value = 0.1065 If AGE has no effect overall, there is a 11% chance of randomly observing an effect this big (or bigger) in an experiment of this size. The effect is considered not significant. Importantly serotonin is present in the EC cells and is responsible in the driving of the colonic motility. After investigating is serotonin is affected by age, it was evident that availability of serotonin influences the role of melatonin. There was a significant age related decline in the MEL/serotonin ratio. Further analysis shows that MEL /serotonin decreased considerably between 3 and 18 months. These observations suggest that MEL may be the mucosal signaling molecule Stroking or applying pressure on the mucosa or the circumferential periphery, always generate CMMC similar to identical spontaneous CMMCs. These generated CMMCs are usually more synchronised and transit smoothly over the entire colon length. However, a graded response to response is not identical for all the ages tested. Upon addition of ondansetron, to the faecal pellet, it significantly decreased the CMMc amplitude and the propulsive tension on the pellet by around 75 percent as shown in figure 1 above. This suggests the importance of circumferential stretch in initiating the CMMC. Electrical field stimulation of distal colon with and without mucosa Simultaneous microelectrode recordings of the intercellular were conducted on distal colon with and without mucosa was connected with rope to platinum transmural stimulating wire, which was connected to a Grass SD stimulator (MA). This was done on both oral and anal side of the selected colon. The tabulated results were as shown in figure 2 below. Regular CMMCs were observed consisted of hyperpolarization proceeding depolarization, from which superimposed excitatory junction potentials were created. It was determined that CMMc started in oral area moving all the way to the rectum. Inhibitory junction potentials were recorded in between the CMMCs. Discussion From the above results, it is apparent that there exists a synergetic relationship between the local reflexes and spontaneous CMMCs the propagation of faecal pellet down is mostly dependent on the CMMC, which mostly occurs involuntary within an empty colon. However, the presence of colon contents determine the location of origin within the colonic CMMC and its propagation down the colon through the activation of both mucosal and circumferential reflexes. The fact that faecal pellet organizes CMMC confirms that a pellet in the colon exerts a dominant effect on the neutral circuitry tasked with generating CMMCs. The role of 5-HT3 receptors From the results in the experiment, we found that ondansetron substantially minimised the responses to the mechanical stimulation of the mucosa using the artists’ brush, rather not the responses of the circumferential stretch, thereby significantly decreasing the propulsive tension on the amplitude of the CMMC over the pellet in the colon. It is evident from the illustrations that transit time within the proximal and distal area due to the reduction in the speed and frequency of the stepwise traversed and the displacement covered in every stepwise. It is also evident that, despite the already verified information on colon motility available, the nervous circuitry mediating the circular and longitudinal muscles is yet to be fully understood. The enteric pathways create overlapping networks called local modules whose interaction determines the generation of motor patterns. The contribution of the effect of age on the function of colon leading to disorder and constipation in human is incompletely understood. Accordingly numerous recent studies have shown 5-hydroxytryptamine (5HT) is to be essential for initiating reflexes resulting from mucosal stimulation and also for normal propagation of colonic migrating motor complexes (CMMCs) together with propulsion of faecal pellets in isolated colon. Evident changes with age occurrence spontaneously in human such as colonic dysmotility and constipation have raised the question about the effect of ageing on enterochromaffin (EC) cells, which may suggest age related issue in function of these cells. The results concurs with the understanding that ondasentron almost completely the mucosal stimulation response as observed from the results. From these results it is evident that 5-HT excreted from EC cells within Mucosa is crucial in initiating mucosal compression of the generated by stroking the mucosa. The effects are more pronounced in the 24 month old mice as compared with the other groups of tested mice. Furthermore this study clarifies the role performed by change in 5-HT release properties with age by informing of the likely changes in humans and impact on human health in relations of understanding the physiological and pathological changes and influences therapeutic approaches to some of the conditions and facilitating in an increase in more study within these area of research. Simultaneous motor responses of the circular and longitudinal showed in distal and proximal segments of the colon proved that inhibitory and excitatory reflex patterns in the ascending contraction in both longitudinal and circular muscle within descending relaxation experienced in the circular muscles can be activated via the same stimuli. Conclusion This study clarifies the role performed by change in 5-HT release properties with age by informing of the likely changes in humans and impact on human health in relations of understanding the physiological and pathological changes and influences therapeutic approaches to some of the conditions and facilitating in an increase in more study within these area of research. The interaction between colonic mucosal regeneration and ageing has been concluded by a number of studies, however such results only provide as short insight into the pathologic and physiologic mucosal healing within the ageing context. An understanding of the impact of the age- related mechanisms may assist in the rising new therapeutic approaches for chronic colonic diseases accompanied with mucosal destruction. Works Cited Alvarez , W. C., 1984. An Introduction to Gastroenterology. 4 ed. New York: Paul B. Hoeber. Bassotti , G. & Villanacci , V., 2006. 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