Effect of Acetaminophen Sulfate on the Growth of Neurite in PC 12 Cells under Various Concentrations (Lab Report Sample)

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Effect of Acetaminophen Sulfate on the Growth of Neurite in PC 12 Cells under Various Concentrations
Summary
Background: Neurite outgrowth is the process by which the nerve cells undergo a development process to produce new projects in neuron cells which can result in increased activities of the neurons in the brain. Acetaminophen sulfate is an inactive metabolite of acetaminophen (paracetamol), a low therapeutic drug which is widely used to control pain and reduce fever.
Materials and Methods: The experiment presumes that different concentrations of acetaminophen sulfate can influence the neurite outgrowth in PC 12 cells lines derived from the  HYPERLINK "https://en.wikipedia.org/wiki/Pheochromocytoma" \o "Pheochromocytoma"
Pheochromocytoma in the medulla of a rat. Different concentrations of acetaminophen sulfate were mixed with NGF-induced PC12 cells to test whether or not acetaminophen sulfate would affect neurites outgrowth. The test also analyzed the rate of the neurite outgrowth among the different concentration of acetaminophen sulfate.
Results: With acetaminophen sulfate treatment at varying concentrations, there was no effect on the growth of neurites on the NGF primed PC12 cells.
Conclusion: The results are similar to previous observations from in-vitro studies which concluded that acetaminophen sulfate does not affect neurite outgrowth in PC12 cells.
Introduction
Neurons are the basic functional units of the nervous system that receive and communicate information in the brain by transmitting electrical impulses (signals) from one part of the brain to another, the sensory organs to the spinal cord and from the spinal cord to the receptor organs. There are over 100 billion nerve cells in the brain which communicate with each other in an organized manner to form a neuron network (Byrne np). A single neuron is made up of three parts, the dendrite, the axon and the cell body. The dendrites receive input information from sensory organs and other neurons. The cell body interprets the information and sends an output signal through the axon to the effector organs and other neurons (Cherry np). Electrical impulses from the axon stimulate the release of specialized chemical signals called neurotransmitters such as the endorphins, epinephrine, and acetylcholine. These neurotransmitters affect several physiological functions such metabolism, learning, sleep, appetite and movement.
Neurons can undergo a development process known as neurite outgrowth to produce new projections which can mature into dendrites or axons. The development of the neurites increases the neuron network and the activities of the nervous system in the brain. The inhibition of the neurite outgrowth can result in impaired cognitive functions (Meldoles np). The neurite outgrowth can be stimulated by several factors such as nerve growth factors and neurotrophins. Medications and pharmacological ingredients can also affect neurite outgrowth. Understanding the physiology and mechanism of neurite outgrowth can help to shed more light on the biology of some neurodegenerative diseases. Scientists apply several special techniques to grow neuron cells using specialized cell lines such the PC12 cells, stimulate neurite growth using neurogenic compounds and monitor the behavior of the growing neurites (Millecamps & Jean-Pierre np).
PC 12 cells are used to assess how varying concentration so of acetaminophen sulfate affects the outgrowth of neurites. PC 12 cells are cell lines that are extracted from the  HYPERLINK "https://en.wikipedia.org/wiki/Pheochromocytoma" \o "Pheochromocytoma"
pheochromocytoma of the rat.  HYPERLINK "https://en.wikipedia.org/wiki/Pheochromocytoma" \o "Pheochromocytoma"
Pheochromocytoma is a malignant tumor in the medulla of the adrenal glands or the chromaffin tissue which produces and stores a lot of catecholamines such as norepinephrine and epinephrine. PC12 cells are small and irregular scattered cells which grow in suspension or liquid media (Wang et al. 62).
The PC12 cells originate from the neural crest which has both the primitive nerve cells and eosinophilic cells. The embryonic nature of the PC12 cells means that they can be cultured indefinitely and can readily differentiate into cells that share the same properties as the nerve cells such as the release of neurotransmitters (Mingorance-Le, Alma & Timothy np). Immature and undifferentiated PC12 cells have no neurites and have a little responsive capacity to neurotransmitters.
When exposed to nerve growth factor (NGF), the PC12 cells stop the replication process and start to differentiate into irregular neuron-like cells with many long projections similar to those seen in neurites. NGF is essential in regulating the growth and differentiation of the neurons, and the performance of the central and peripheral nerve cells. The differentiated PC12 cells can be excited by electrical impulses and respond to neurotransmitters (Oliveira et al. 78).
Because of the similarities between the differentiated PC12 cells and the nerve cells, PC12 cells have extensive application in the study neurological diseases by creating artificial tissues of the nervous, which can be used to study neurodegenerative and neurochemicals. PC12 cells are preferred for neurological studies such as the NGF-induced neurite formation over other cells lines because of their unique abilities to exit the cell division cycle and differentiate into other cells types when exposed to certain chemicals. Unlike the chromaffin cell lines, PC12 cell lines can also proliferate easily in growth media and release hormones such as catecholamines upon stimulation (Malagelada & Lloyd 376).
In cell culture, the PC12 cell lines are grown in suspension media which contains amino acids, serum, glucose, vitamins, and antibiotics such as penicillin to prevent the contamination of the cell culture by bacteria and fungi. The media also constrains a red indicator which turns to yellow when the cells are alive and purple when the cells are dead.
Laminin is a glycoprotein that forms part of the extracellular matrix in human and animals. The extracellular matrix is located outside the cells and is important in providing support and attachment to the cells in the tissue and organs. The structure of laminin is vital to its functions. Laminin is a cross-linked molecule that is made up of a combination of alpha, beta and gamma chains located on three short extensions and one long extension (Aumailley 48).
The extensions on the laminin are essential in cell adhesion. The short extensions of the laminin molecules allow them to bind to each other to form long sheets while the long extension can attach to cells and anchor them to the plasma membrane. Currently, there are more than 16 laminin molecules which are important in gluing cells to the connective tissues and promote normal cellular functions (Domogatskaya, Sergey & Karl np).
The function of laminin in the body is to facilitate cellular attachment and enhance motility, proliferation, and differentiation of cells. Laminin binds to cell membranes using the integrin receptors and other glycoprotein molecules such as dystroglycan (Yurchenco 5-7). This ensures the survival of tissues and that the shape of the cells and the structure of the tissues are maintained efficiently. Laminin glycoprotein is secreted outside the cells and incorporated into the extracellular matrix of the cells.
Laminin is used to enhance cell cultures and provide a conducive environment for the neuron cells, cardiomyocytes and endothelial cells to adhere, proliferate and differentiate in-vitro. Commercially available laminin are extracted from mouse sarcoma (Laminin-111) and human placenta (laminin-211, 411 or 511). Laminin-111 serves as a substrate for the growth of nerve cells in the body and cell cultures. In vitro cells, studies have demonstrated the importance of laminin in enhancing cell adhesion, neurite growth and survival of cells under stress (Miyazaki et al. 7).
Impaired laminin can result in the improper formation of tissues and organs such as the brain, the kidney, and the skin, resulting in muscular dystrophy and nephrotic syndrome. Some laminin has also been linked to the spread and invasive nature of malignant cancers. A recent human study has reported that over-expression of the laminin-511 was critical in the dissemination and invasion of tissue in patients with advanced breast carcinoma. Therefore, by targeting laminin-511, scientists can develop therapeutic strategies in cancer patients (Pouliot & Nicole 142-143).
Acetaminophen sulfate is a pharmaceutical metabolite of acetaminophen (paracetamol). Acetaminophen is commonly used medicine for the management of pain and fever. It is a commonly prescribe over the shelf medication in adults and children and has a low therapeutic index. Acetaminophen inhibits the production of prostaglandins in the tissues during injuries, by inhibiting the activities of cyclooxygenase 2 (COX) enzymes (Hinz & Kay 20).
Prostaglandins are a group of lipid hormones that are involved in the healing processes, resulting in inflammation, pain, and fever. By reducing the synthesis of prostaglandins, the pain threshold in the body and the temperature control center are regulated, thus reducing pain and fever (Ricciotti & Garret 987). In Humans, mice, and rats, acetaminophen is metabolized in the liver and converted to inactive substances which are eliminated from the body through the urine. In the liver, the cytochrome P450 enzyme system metabolizes a small concentration of acetaminophen. Acetaminophen is oxidized to produce a reactive and toxic intermediary metabolite N-acetyl-p-benzoquinone imine (NAPQI). The toxic acetaminophen compound combines with the sulfhydryl group and is converted t...