This essay SF-9 Lepidopteran Cells has a total of 3244 words and 16 pages.
SF-9 Lepidopteran Cells
PHM499 Research Project
Supervisors: Dr. P. S. Pennefather, Dr. S. M. Ross
Calcium transport study of SF-9 lepidopteran cells and bull frog sympathetic ganglion cells
Faculty of Pharmacy, University of Toronto, 19 Russell Street, Toronto, Ontario M5S 2S2
ABSTRACT The intracellular calcium level and the calcium efflux of the bull-frog sympathetic ganglion cells (BSG) and the SF-9 lepidopteran ovarian cells were investigated using a calcium-sensitive fluorescence probe fura-2. It was found that the intracellular calcium levels were 58.2 and 44.7 nM for the BSG cells and SF-9 cells respectively. The calcium effluxes following zero calcium solution were 2.02 and 1.33 fmole·cm-2·s-1 for the BSG cells and SF-9 cells. The calcium effluxes following sodium orthovanadate (Na2VO4) in zero calcium solution were 6.00 and 0.80 fmole·cm-2·s-1 for the BSG cells and the SF-9 cells. The SF-9 cells also lost the ability to extrude intracellular calcium after 2-3 applications of Na2VO4 while the BSG cells showed no apparent lost of calcium extruding abilities for up to 4 applications of Na2VO4.
Spodoptera frugiperda clone 9 (SF-9) cells are a cultured insect cell line derived from the butterfly ovarian tissue. SF-9 cells are used by molecular biologists for the studies of gene expression and protein processing (Luckow and Summers, 1988). However, there is not much known about these cells\' basic biophysiology. Since calcium is involved in many cells\' activities such as acting as a secondary messenger, it is important for cells to control their intracellular calcium level. This study was aimed toward looking at the some of the basic properties of the SF-9 cells such as resting calcium concentration and rate of calcium extrusion after being calcium level being raised by an ionophore 4-bromo-A23187. The effect of sodium orthovanadate (an active transport inhibitor) on calcium extrusion was also looked at. Microspectrofluorescence techniques and the calcium-sensitive probe fura-2 were used to measure the intracellular calcium concentration of these cells. In addition, the BSG cells were used to compare with the SF-9 cells for the parameters that were studied.
It was found that the SF-9 cells appeared to have a calcium concentration similar to the BSG cells. Moreover, the calcium extrusion rates of both cell types with no Na2VO4 added seemed to the same. However, due to insufficient data, the effects of Na2VO4 could not be statistically analyzed. From the data available, it suggested that the BSG cells\' rate of calcium extrusion was enhanced by the Na2VO4 and was greater than the SF-9 cells. It was more important to note that the calcium extruding capabilities of the SF-9 cell seemed to impaired after two to three applications of Na2VO4 but it had apparent effects on the BSG cells even up to 4 applications.
After obtaining these basic parameters, many questions raised such as how does the SF-9 cells extrude their calcium and why the Na2VO4 affected the calcium efflux for the SF-9 cells but not the BSG cells? The SF-9 cells may have a calcium pump or exchanger to extrude their calcium and they may be very sensitive to the ATP (adenosine 3\'-triphosphate) supply. This was apparently different from the BSG cells\' since their calcium extrusion were not affected by the Na2VO4.. It may be useful to find the mechanism(s) of the actions of Na2VO4 on the SF-9 cells because it may find possible applications in agriculture such as pest control.
MATERIALS AND METHODS
Chemicals and solutions
4-bromo-A23187 and Fura-2/AM were purchased from Molecular Probes (Eugene, OR). Na2VO4 was purchased from Alomone Lab (Jerusalem, Israel). Dimethyl sulfoxide (DMSO) was obtained from J. T. Baker Inc. (Phillipsburg, NJ). All other reagents were obtained from Sigma (St. Louis, MO).
The normal Ringer\'s solution (NRS) contained (mM): 125 NaCl, 5.0 KCl, 2.0 CaCl2, 1.0 MgSO4, 10.0 glucose, 10.0 N-[2-hydroxyethyl] piperazine-N\'-[2-ethanesulfonic acid] (HEPES). The calcium free Ringer solution (0CaNRS) is the same as the NRS except CaCl2 was substituted with 2.0 mM ethylene glycol-bis(b-aminoehtyl) ether N,N,N\',N\'-tetraacetic acid (EGTA).
Fura-2/AM solution was prepared as follows: a stock solution of 1mM fura-2/AM in DMSO was diluted 1:500 in NRS containing 2% bovine albumin. It was then sonicated for 10 minutes. It was then kept frozen until the day of the experiment.
20 SYMBOL 109 f "Symbol"M 4-bromo-A23187 solution was prepared by diluting a stock of 5mM 4-bromo-A23187 in DMSO 1:250 with NRS.
Na2VO4 solution (VO4NRS) contained 100 SYMBOL 109 f "Symbol"M. Na2VO4 in 0CaNRS.
Alexander The GreatAlexander The Great Alexander the Great and His Achievements Alexander the Great was the king of Macedon. Alexander of Macedon, or ancient Mecadonia, deserves to be called the Great. Alexander the Great was considered one of the greatest military geniuses of all times. He was an excellent king, general, and conqueror. During his thirteen-year rule he conquered almost all the then known world and gave a new direction to history. He had established an empire after he died. His new empire helped ma
PantheonPantheon Introduction I chose to report on the pantheon because I’ve seen pictures and I am also very fascinated by all the Roman monuments. I looked all around the internet and libraries. The Pantheon was very hard to find information about. It was very challenging but I found enough information to complete this report. This famous building stands in the business district of Romemuch as it was built some 18 centuries ago. Amazingly, it has withstood the ravages of both the elements and war perm
The PantheonThe Pantheon Introduction I chose to report on the pantheon because I’ve seen pictures and I am also very fascinated by all the Roman monuments. I looked all around the internet and libraries. The Pantheon was very hard to find information about. It was very challenging but I found enough information to complete this report. This famous building stands in the business district of Romemuch as it was built some 18 centuries ago. Amazingly, it has withstood the ravages of both the elements and war
THE EFFECTS OF ALTITUDE ON HUMAN PHYSIOLOGYTHE EFFECTS OF ALTITUDE ON HUMAN PHYSIOLOGY Changes in altitude have a profound effect on the human body. The body attempts to maintain a state of homeostasis or balance to ensure the optimal operating environment for its complex chemical systems. Any change from this homeostasis is a change away from the optimal operating environment. The body attempts to correct this imbalance. One such imbalance is the effect of increasing altitude on the body\'s ability to provide adequate oxygen to be utili
FrankensteinFrankenstein In the story “Frankenstein,” written by the author Mary Shelley, Victor Frankenstein decided that wanted to create a being out of people that were already dead. He believed that he could bring people back from the grave. Playing with nature in such a way would make him play the role of God. With Victor Frankenstein feeling that he had no true friends, the only relief he had of expressing his feeling was through letters to Elizabeth. Elizabeth was not Victors’ true sister but he love
FrankensteinFrankenstein Victor Frankenstein was very interested in natural philosophy and chemistry and basically tried to play G-d by creating life. When he found the secret of activating dead flesh, he created a superhuman being composed of rotted corpses. What he did was considered unthinkable, and he was haunted by his own creation. When the monster escaped, Frankenstein knew that he had to deal with the consequences of what the monster might do. Frankenstein received a letter one day which informed hi
Ai - Edge Of ExcellenceAi - Edge Of Excellence Running head: The Edge of Excellence The Edge of Excellence Kathleen P. Munn Community College of Philadelphia Recently, the media has spent an increasing amount of broadcast time on new technology. The focus of high-tech media has been aimed at the flurry of advances concerning artificial intelligence (AI). What is artificial intelligence and what is the media talking about? Are these technologies beneficial to our society or mere novelties among business and marketing p
Why The Wall Street Bubble Must Burst?Why The Wall Street Bubble Must Burst? Why the Wall Street Bubble Must Burst? In 1938, and in the teeth of the longest and fiercest depression that the United States had ever known, capital spending hit an all time high. Thats right! In 1938 the men who owned America began to pour millions of Dollars into new plant and equipment as if there was no tomorrow. We dont think much about it today, because it has been a long time since the United States has experienced a real bone jolting economic sl
DuPont An investment analysisDuPont An investment analysis DuPont makes a variety of high-value products for industry today, including polymers, chemicals, fibers, and petroleum products...products for agriculture, electronics, transportation, apparel, food, aerospace, construction, and health care. DuPont serves customers in these and other industries every day, offering better things for better living as the company prepares to begin its third century of scientific, technological, commercial, and social achievement. DuP
LeadLead subject = Chemistry title = Lead Lead is a lustrous, silvery metal that tarnishes in the presence of air and becomes a dull bluish gray. Soft and flexible, it has a low melting point (327 °C). Its chemical symbol, Pb, is from plumbum, the Latin word for waterworks, because of lead\'s extensive use in ancient water pipes. Itsatomic number is 82; its atomic weight is 207.19. Lead and lead compounds can be highly toxic when eaten or inhaled. Although lead is absorbed very slowly into the body,
Chemistry: Water PollutionChemistry: Water Pollution Phosphates in Water Pollution Phosphates may be created by substituting some or all of the hydrogen of a phosphoric acid by metals. Depending on the number of hydrogen atoms that are replaced, the resulting compound is described as a primary, secondary or tertiary phosphate. Primary and secondary phosphates contain hydrogen and are acid salts. Secondary and tertiary phosphates, with the exception of those of sodium, potassium and ammonium are insoluble in water. Tertia
PlatinumPlatinum subject = Chemistry title = Platinum Platinum is a relatively rare, chemically inert, metallic element. It symbol is Pt, atomic number is 78, and its atomic weight is 195.09. Platinum is one of the heaviest substances known. One cubic foot of Platinum weighs 21 times as much as a cubic foot of water. A grayish-white metal, Platinum has a melting point of 1772 degrees C and a realatively high boiling point of 3827 degrees C. It has a high fusing point, is ductile and malleable, expands s
The Chemistry of Natural WaterThe Chemistry of Natural Water INTRODUCTION The purpose of this experiment is to explore the hardness of the water on campus. Hard water has been a problem for hundreds of years. One of the earliest references to the hardness or softness of water is in Hippocrates discourse on water quality in Fifth century B.C. Hard water causes many problems in both in the household and in the industrial world. One of the largest problems with hard water is that it tends to leave a residue when it evaporates.
Artificial IntelligenceArtificial Intelligence Recently, the media has spent an increasing amount of broadcast time on new technology. The focus of high-tech media has been aimed at the flurry of advances concerning artificial intelligence (AI). What is artificial intelligence and what is the media talking about? Are these technologies beneficial to our society or mere novelties among business and marketing professionals? Medical facilities, police departments, and manufacturing plants have all been changed by AI but
Civil EngineeringCivil Engineering Civil Engineering Why do I want to be a civil engineer? Until recently, I did not know the answer to this question myself. I was lost when choosing a career. Then, I read about civil engineering, an occupation involving the construction of buildings, roads, and bridges. As I looked farther into civil engineering, I liked many of the other aspects involved with the career. Although the education will be difficult, I have determined that civil engineering is the career that I wan
FrankensteinFrankenstein This is an English book report on Frankenstein by Mary Shelley Frankenstein The story of Frankenstein by Mary Shelley is about a man who created something that messes with nature, and nature came back to mess with him because nature is more powerful than man. Victor Frankenstein was very interested in natural philosophy and chemistry and basically tried to play G-d by creating life. When he found the secret of activating dead flesh, he created a superhuman being composed of rotted
Isaac AsimovIsaac Asimov Isaac Asimov was born on January 2, 1920 in Petrouchi, Russia. His parents were Judah and Anna Asimov. Isaac also has a sister Veronica and a brother Stanley. In 1923 his family immigrated to the United States. He and his family grew up in Brooklyn, New York. In Brooklyn his family ran a small candy and magazine store. This is one of the places where Asimov began to learn about printing. Also it was here that Asimov learned good business and self-discipline skills (Bloom, 251). Asim
Science Fiction Book ReportScience Fiction Book Report In the story “Frankenstein,” written by the author Mary Shelley, Victor Frankenstein decided that wanted to create a being out of people that were already dead. He believed that he could bring people back from the grave. Playing with nature in such a way would make him play the role of God. With Victor Frankenstein feeling that he had no true friends, the only relief he had of expressing his feeling was through letters to Elizabeth. Elizabeth was not Victors’ true sis
Frankenstein Support (Mock-Persausive Letter FormaFrankenstein Support (Mock-Persausive Letter Format) (Author\'s Note: This was a semi-creative project. We had to address the issues in a persuasive letter rather than a boring ol\' report, so please become unconfused as far as the format..) Cal Tech Curriculum Committee: Scientists are all too ready to lock themselves away with their research, unwilling - perhaps even incapable - of seeing the consequences of their actions. It is our duty as their educators to provide them with not only a means
Plate TectonicsPlate Tectonics Plate Tectonics Since the beginning of human kind there has been a cloud of wonder regarding the origin of our planet. Scientist interested in this field through out the years have developed many different theories to how our planet came about. Before the Twentieth Century, scientist and geologist alike revolved in the idea that Mountain building was due to the massive contractions of the earth caused by the gradual cooling of molten rocks. In 1900, American scientist Joseph Le C
Myth Or Science?Myth Or Science? Throughout history there have been many attempts to explain the origin and workings of our universe. Most every culture has their own cosmogony. Nearly every individual has his or her own idea of what our universe is. During our modern era of advanced scientific knowledge, we feel that we have a good grasp on how the universe works. We have our Chemistry and Physics, along with Mathematics, to examine the universe with. Any person educated in these fields will tell you that they
Bio OutlineBio Outline BIOLOGY 220 OUTLINE SECTION II Text: Essential Cell Biology I. Opening Comments (Chapter 3) A. Life creates order out of disorder through a never-ending series of chemical reactions B. This is Metabolism and the ability to Metabolize C. Most of the chemical reactions required by the cell would not occur at physiological conditions D. Control of these reactions is achieved by specialized protein, ENZYMES. II. Basic Principles of Energy A. Energy - Basics Principles 1. Define Energy -
Construction Of A GeneticisConstruction Of A Geneticis The Construction of a Geneticist I. A. The construction of a geneticist follows a difficult path of education, preparation, and hard work. B. Genetic Engineering C. Scope 1- accomplishments 2- job description 3- education 4- opportunities 5- future 6- (interview) - Personal insight - Attraction to job II. Background III. Accomplishments IV. Job Description & Opportunities V. Education & salary VI. Interview -Attraction, Personal Insight -Future VII. Conclusion- Why I
Air QualityAir Quality Air Quality & Dispersion Today, the air quality aspect of ARL research is by far the dominant theme, but distinctions among the themes remain somewhat vague. For example, the models developed for emergency response purposes are among those used for air quality prediction. The Air Quality and Dispersion theme is one of the strongest ties that binds ARL\'s components together. ARL is not heavily involved in the pure science of the business. Instead, ARL focusses on the need to assemble
AlchemyAlchemy Alchemy There are many ways to examine the subject of alchemy, including alchemy as a source of symbolism, psychology, and mysticism. It has also been an influence on the world view of various writers, artist, and musicians. The focus of this report is alchemy as a pre-chemistry, which gave a new impulse towards the preparation of medicinal remedies and also was a major influence on today\'s scientific investigations. Alchemy is an ancient art, practiced in the Middle Ages. The fundament
History Of ChemistryHistory Of Chemistry History of Chemistry Introduction: Humans have always been very curios creatures. The have always wondered about what they are and why they are here. Our limited knowledge of the environment has always urged for new things to be discovered. The desire to understand the world better has made people search for rational answers, for principles and laws. For centuries people have tried to unlock the mysterious world that surrounds them. History: Because myths did not explain thi
HormonesHormones IntroductionHormones are organic substances that are secreted by plants and animals and that function in the regulation of physiological activities and in maintaining homeostasis. They carry out their functions by evoking responses from specific organs or tissues that are adapted to react to minute quantities of them. The classical view of hormones is that they are transmitted to their targets in the bloodstream after discharge from the glands that secrete them. This mode of discharge (
InterviewInterview The Interview For my interview, I interviewed Jennifer Wong, a scientist in the development of analytic chemistry. In her occupation she purifies and analyzes proteins, as well as characterizes the proteins. She works for Panox located at 10301 Stellink Houston, TX 77025. For this company Dr. Wong analyzes proteins in order for her company to market drugs that will later help patients with certain diseases. Right now her company has not yet marketed any products yet, but they are still
Tromaville CoalitionTromaville Coalition Member of the Tromaville Coalition Chemistry 1. a) Bulk movement is the overall movement of a fluid. The molecules all move in the same direction. Diffusion however is the random movement of molecules which usually results in a fairly even distribution. In other words the movement is not guaranteed to move in one direction but the probability that it will move in the lower gradient is greater. Osmosis is similar to diffusion but is differentiated by the membrane\'s behavior.
The Greatest Scientific FraudThe Greatest Scientific Fraud Miriah Killam Writing 122 David Rothgery October 12, 2000 The Greatest Case of Scientific Fraud I have been told I have a beautiful smile, and I once thought it was true. It is big and quick, the kind that flashes across a whole face to reveal pearly whites. The sad truth is, I have become slower to show my Colgate smile as I have watched the most important part of a smile, my teeth, become less and less dazzling. For years I couldn\'t understand; I brushed twice a