To analyze the Glycogen and Protein concentration of a rat liver cell sample, utilizing Iodine solution, Diphenylamine reagent, Biuret reagent and by using heat through boiling.
Introduction
This experiment was designed to exemplify and cipher the distribution of RNA, DNA, protein and Glycogen between the karyon and cytolplasim of the liver cell. The liver is the largest splanchnic organ in the organic structure and signifiers about 3-5 % of the organic structure weight. It likely originated as a digestive secretory organ but is now besides used involved in elimination, storage, homeostasis and the care of consistence in the internal environment of the organic structure. The liver is present in lancelet and craniates, but the construction called the liver in some invertebrates, such as gasteroids, has few belongingss of the vertebraic liver. ( W. H. Horner Andrews, 1979 )
In this experiment rat liver cells were used. There are many differences between an animate being and works cell. Plant cells are surrounded by a midst cell wall for protection which contains cellulose. This helps the works cells to let high force per unit area to construct inside of it, without spliting. A works cell has to be able to accept big sums of liquid through osmosis, without being destroyed. An carnal cell does non hold a cell wall in their construction. If you were to make full the animate being cell with excessively much fluid, it will finally split. ( Wise Geek, 2010 ) Besides works cells, when viewed under the microscope, appear highly different than an animate being cell because of the presence of a big vacuole, which exists in the cell s cytol. It normally takes up most of the room in the cell, and the membrane of the cell encircles it. It contains waste stuffs, H2O, and foods that can be used or secreted as necessary. There are besides many other constituents present in the animate being cell, such as ribosome s, chondriosome and lipid droplets. ( Wise Geek, 2010 )
In every cell nucleus in the organic structure there is DNA within the cell nucleus. The Deoxyribonucleic acid s chief functional function is to transport the cell s information on strands of cistrons, which are found along chromosomes. Ribonucleic acid or RNA is besides present in the liver cell. RNA is one of the three major supermolecules ( along with DNA and proteins ) that are indispensable for all signifiers of life. RNA is made up of constituents called bases. The sequence of bases allows RNA to encode familial information. There are different types of RNA, such as courier RNA, ribosomal RNA and Transfer RNA. ( C. E. Ophardt, 2003 ) These all carry out different maps to reassign and form information and codifications within the cell. Glycogen is besides present in the liver cell. Glycogen is found as a polyose and is derived from the saccharide group. Its chief map is to be readily converted to glucose as needed by the organic structure to fulfill its energy demands. ( Wise Geek, 2010 ) Proteins are besides found within the liver cells. Proteins are organic compounds made of aminic acids arranged in a additive concatenation and folded into a ball-shaped signifier. The liver is0 non a protein shop but it is extremely active in protein metamorphosis.
Consequences
Trial for Glycogen
Test tubing containing:
Test NP and CP for DNA
Test Tube Containinglight brown
Test NP and CP for Protein
Test tubing incorporating
Test NS2 and CS2 for RNA fragments
Test tubing incorporating
Quantitative Assay of Protein
Test tubing figure
CP = 10.5 ( milligram )
Table 1-5: The optical density degrees of 10 trial tubings incorporating different solutions after 30 proceedingss at room temperature, as measured against a clean sample in the spectrophotometer at a wavelength of 550nm. The big difference in values for NP and CP, shown in the text box beside the tabular array, implies inaccurate pippeting.
Appendix 1:
The affiliated standardization curve plots the A550 measurings against protein content. The curve is good shaped, reflecting appropriate and accurate consequences obtained.
Discussion
The liver is a big, soft organ which alters in size and form harmonizing to the sum of blood nowadays. Cholesterol may be found in big measures in the liver. Bile is the chief vehicle of cholesterin elimination. Cholesterol may be secreted to transport out several maps, portion of its synthesis is to replace the cholesterin lost from the membrane when the gall salts are secreted. It is besides used to organize micelles in the gall. Phospholipids are besides found in the gall. Again, they perchance may hold been torn out of the membrane when gall salts are secreted and it is involved in micelle formation. ( W. H. Horner Andrews, 1979 )
Within the cytolpasim of the liver cell many cell organs can be found. These include mitochronria for cell respiration and energy formation. The endoplasmic Reticulum ( ER ) is an eucaryotic cell organ that forms an interrelated web of tubules, cysts, and cisternae within cells. The endoplasmic Reticulum has two parts to it. The Rough ER has many Ribosomes run alonging its interior and creates proteins for the cell which are either used or sent to the Golgi Apparatus for transit. Proteins that are created in the Rough ER have a higher opportunity of being secreted. The smooth ER ‘s map is to take toxins from the system, which is why a high concentration of Smooth ER is found in the liver. Lysosomes are besides found in the liver cell. A Lysosome is an cell organ found in all types of animate being cells which contains a big scope of digestive enzymes capable of dividing most biological supermolecules. The Golgi Apparatus is an cell organ found in most eucaryotic cells, including those of workss, animate beings, and fungi. This cell organ ‘s primary map is to treat proteins targeted to the plasma membrane, lysosomes or endosomes and those that will be formed from the cell, and screen them within cysts. Therefore, it functions as a cardinal bringing system for the cell. ( M. W. King, 2008 ) Peroxisomes are a cell organelle incorporating enzymes, such as catalase and oxidase, that catalyze the production and dislocation of H peroxide. ( The Medical Dictionary, 2010 ) There are besides lipid drolets present in the liver cells cytol. Ribosomes are present in the macromolecular composite but are non classified as cell organs.
The concentration of Sodium and of Potassium is similar in both gall and plasma. This is likely due to diffusion through, or between, cells of the bilious epithelial tissue. The existent and comparative concentration of ion may change with the secretory province of the liver. When secernment of hydrogen carbonate is stimulated, there is a autumn in chloride concentration. With a stimulation of flow with bile salts, there is normally a autumn in bicarbonate concentration, though there is an addition of out put per minute. ( W. H. Horner Andrews, 1979 ) Possible reading of this is that usually there is an equilibrium between plasma and gall, so when gall flow additions there is a greater concentration gradient, favoring a faster transition of hydrogen carbonate into gall. ( W. H. Horner Andrews, 1979 )
The organic structure stored saccharides as Glycogen, a compound formed by the polymerization of glucose. Other hexoses, such as brain sugar and fruit sugar, are converted into glucose by the liver before being stored. Storage in the liver is aided by insulin, the endocrine from the beta cells of the pancreas, but is non indispensable when the concentration of sugar in the blood is high. Up to 10 % of the liver weight ( wet ) may be animal starch, but 3-5 % is more common, about half of the organic structures shop of animal starch is in the liver. Enzymes in the liver may interrupt down animal starch ( glycogenesis ) with great celerity and animal starch disappears shortly after decease unless stairss are taken to suppress the enzymes. Adrenaline, norepinephrine and the stimulation of the sympathic nervousnesss induce the dislocation of animal starch by the hepatocytes and a sudden fear produces a rush of glucose into the blood. During violent muscular excercise, the animal starch of the musculuss is broken down into glucose and in bend, into the lactic acid. This creates the Cori Cycle. The function of the liver is to maintain the concentration of glucose in the blood every bit changeless as possible, the encephalon particularly suffering harm if the concentration falls excessively low. ( W. H. Horner Andrews, 1979 )
Figure 1-1: The construction of Glycogen
M. W. King
Although the liver is non a protein shop, it is extremely active in protein metamorphosis and about all of the proteins in plasma have an hepatic beginning. Proteins are built up from amino acid bases, from nutrient or coming from other tissues. Plasma proteins can be classified into Albumins and Globulins. Plasma contains several bearer proteins, such as transferran for Fe, ceruloplasmin for Cu and yet another to adhere adrenal cartiel endocrines. The blood contains many blood coagulating factors which are proteins synthesised by the liver, besides known as fibrinolysinogen. This can be split to organize fibrinolysin ( plasmin ) , a substance which prevents intravascular curdling. ( W. H. Horner Andrews, 1979 )
Within the liver there are several different types of RNA nowadays. The courier RNA carries the familial codification into the cytol where protein synthesis occurs. Each cistron ( or distinct section ) on Deoxyribonucleic acid contains instructions for doing one specific protein with order of amino acids coded by the precise sequence of heterocyclic aminoalkanes on the bases. It has been determined that the familial codification is really based upon threes of bases which provide 64 different codifications utilizing the four bases. ( C. E. Ophardt, 2003 ) These 4 bases are Adenosine, Thymine, Guanine and Cytosine. Each nucleotide three, called a codon, can be “ read ” and translated into an amino acid to be incorporated into a protein being synthesized. In the cytol, ribsomal RNA ( rRNA ) and protein combine to organize a nucleoprotein called a ribosome. The ribosome serves as the site and carries the enzymes necessary for protein synthesis. There are about equal parts rRNA and protein. The ribosome attaches itself to m-RNA and provides the stabilizing construction to keep all substances in place as the protein is synthesized. Several ribosomes may be attached to a individual RNA at any clip. Transfer RNA ( transfer RNA ) contains about 75 bases, three of which are called anticodons, and one amino acid. The transfer RNA reads the familial codification and carries the amino acid to be incorporated into the developing protein. There are at least 20 different transfer RNA ‘s – 1 for each amino acid. The basic construction of a transfer RNA is shown in the left graphic. Part of the transfer RNA doubles back upon itself to organize several dual coiling subdivisions. On one terminal, the amino acid, phenylalanine, is attached. On the opposite terminal, a specific base three, called the anticodon, is used to really “ read ” the codons on the messenger RNA. ( C. E. Ophardt, 2003 )
Figure 1-2: The construction of a strand of RNA
Astrobiology Magazine, 2008
Decision
In this experiment it was found that Glycogen and Protein were present in a sample of rat liver cells. Their presence can be detected by executing experiments with Iodine solution, Diphenylamine reagent, Biuret reagent and by using heat through boiling. There was inaccurate pipetting during the quantative check of protein, ensuing in a difference in values between NP and CP consequences. It can be concluded that NP is more abundant in a rat liver cell than that of CP.
