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Cytoplasm: The part of a cell that contains the Cytosol and small structures excluding the Cell nucleus; Mitochondria; and large Vacuoles. (Glick, Glossary of Biochemistry and Molecular Biology, 1990)

What is Cellular Respiration?

JoVE 10976

Organisms harvest energy from food, but this energy cannot be directly used by cells. Cells convert the energy stored in nutrients into a more usable form: adenosine triphosphate (ATP).

ATP stores energy in chemical bonds that can be quickly released when needed. Cells produce energy in the form of ATP through the process of cellular respiration. Although much of the energy from cellular respiration is released as heat, some of it is used to make ATP. During cellular respiration, several oxidation-reduction (redox) reactions transfer electrons from organic molecules to other molecules. Here, oxidation refers to electron loss and reduction to electron gain. The electron carriers NAD+ and FAD—and their reduced forms, NADH and FADH2, respectively—are essential for several steps of cellular respiration. Some prokaryotes use anaerobic respiration, which does not require oxygen. Most organisms use aerobic (oxygen-requiring) respiration, which produces much more ATP. Aerobic respiration generates ATP by breaking down glucose and oxygen into carbon dioxide and water. Both aerobic and anaerobic respiration begin with glycolysis, which does not require oxygen. Glycolysis breaks down glucose into pyruvate, yielding ATP. In the absence of oxygen, pyruvate ferments, producing NAD+ for continued glyc

 Core: Biology

Cell-surface Signaling

JoVE 10877

Hormones—or any molecule that binds to a receptor, known as a ligand—that are lipid-insoluble (water-soluble) are not able to diffuse across the cell membrane. In order to be able to affect a cell without entering it, these hormones bind to receptors on the cell membrane. When a first messenger, a hormone, binds to a receptor, a signal cascade is set off, causing second messengers, proteins inside the cell, to become activated, resulting in downstream effects. Cell membrane receptors have three portions: an external ligand-binding domain, a transmembrane domain, and an internal domain. There are three categories of cell membrane receptors based on the consistency of the structure and function of these domains within each category. One category is ligand-gated ion channels which, when bound to a ligand, undergo a conformational change, allowing ions through a channel formed by the transmembrane portion of the receptor. A second category is G-proteins-coupled receptors which have a distinct structure with seven transmembrane domains. Binding of the external domain to a ligand causes the alpha subunit, one of three subunits attached to the internal portion of the receptor, to disassociate from the receptor and create a cellular response. The third category of receptors, the enzyme-linked receptor—also called catalytic receptor

 Core: Biology

Viral Recombination

JoVE 10826

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.

Some diseases can infect multiple species. For example, pigs can be infected by some human and bird viruses, in addition to the viruses that usually infect pigs. Because viruses can recombine when they co-infect the same cell, pigs can act like “mixing vessels” that recombine viruses from other species to create new viruses that can sometimes infect humans. This worrisome phenomenon represents a route through which infectious material from other species can enter the human population. Diseases that move from animals to humans are known as zoonoses. Humans can be highly susceptible to such viruses because we have no history of exposure that would have generated immunity. Influenza A is a prime example of the “mixing vessel” theory of viral disease. Research has demonstrated that pig, bird, and human influenza A viruses have reassorted inside pig hosts. These events yielded “double reassortant” viruses that contained genes from human and bird viruses and “triple

 Core: Biology

Meiosis I

JoVE 10767

Meiosis is a carefully orchestrated set of cell divisions, the goal of which—in humans—is to produce haploid sperm or eggs, each containing half the number of chromosomes present in somatic cells elsewhere in the body. Meiosis I is the first such division, and involves several key steps, among them: condensation of replicated chromosomes in diploid cells; the pairing of homologous chromosomes and their exchange of information; and finally, the separation of homologous chromosomes by a microtubule-based network. This last step segregates homologs between two haploid precursor cells that may subsequently enter the second phase of meiosis, meiosis II. The exchange of equivalent segments between homologous chromosomes occurs early on during meiosis I, and is referred to as crossing over. This process relies on the close association of such homologs, which are drawn together by the formation of a connective protein framework called the synaptonemal complex between them. To function correctly, the complex requires three parts: (1) vertical lateral elements, which form along the inward-facing sides of two juxtaposed homologous chromosomes; (2) a vertical central element positioned between the chromosomes; and (3) transverse filaments, or horizontal protein threads that connect the vertical and central components. The result has often been compared to a ladde

 Core: Biology

Electron Carriers

JoVE 10744

Electron carriers can be thought of as electron shuttles. These compounds can easily accept electrons (i.e., be reduced) or lose them (i.e., be oxidized). They, therefore, play an essential role in energy production because cellular respiration is contingent on the flow of electrons.

Over the many stages of cellular respiration, glucose breaks down into carbon dioxide and water. Electron carriers pick up electrons lost by glucose in these reactions, temporarily store the electrons and input them into the electron transport chain. Two such electron carriers are NAD+ and FAD, which are both derived from B vitamins. The reduced forms of NAD+ and FAD, NADH and FADH2, respectively, are produced during earlier stages of cellular respiration (glycolysis, pyruvate oxidation, and the citric acid cycle). The reduced electron carriers NADH and FADH2 pass electrons into complexes I and II of the electron transport chain, respectively. In the process, they are oxidized to form NAD+ and FAD. Additional electron carriers in the electron transport chain are flavoproteins, iron-sulfur clusters, quinones, and cytochromes. With the assistance of enzymes, these electron carriers eventually transfer the electrons to oxygen molecules. The electron carriers become oxidized as they donate electrons and re

 Core: Biology

G-protein Coupled Receptors

JoVE 10718

G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.

The G-protein or guanine nucleotide-binding protein, is a large heterotrimeric complex. Its three subunits are labeled alpha (α), beta (β), and gamma (γ). When the receptor is unbound or resting, the α-subunit binds a guanosine diphosphate molecule or GDP, and all three subunits are attached to the receptor. When a ligand binds the receptor, the α-subunit releases the GDP and binds a molecule of guanosine triphosphate (GTP). This action releases the α-GTP complex and the β-γ complex from the receptor. The α-GTP can move along the membrane to activate second messenger pathways such as cAMP. However there are different types of α-subunits and some are inhibitory, turning off cAMP. The β-γ complex may interact with potassium ion channels which release potassium (K+) into the extracellular space resulting in hyperpolarization of the cell membrane. This type of ligand-gated ion channel is called a G-prot

 Core: Biology

Protein Associations

JoVE 10704

The cell membrane—or plasma membrane—is an ever-changing landscape. It is described as a fluid mosaic as various macromolecules are embedded in the phospholipid bilayer. Among the macromolecules are proteins. The protein content varies across cell types. For example, mitochondrial inner membranes contain ~76%, while myelin contains ~18% protein content. Individual cells contain many types ofbrane proteins—red blood cells contain over 50—and different cell types harbor distinct membrane protein sets. Membrane proteins have wide-ranging functions. For example, they can be channels or carriers that transport substances, enzymes with metabolic roles, or receptors that bind to chemical messengers. Like membrane lipids, most membrane proteins contain hydrophilic (water-loving) and hydrophobic (water-fearing) regions. The hydrophilic areas are exposed to water-containing solution inside the cell, outside the cell, or both. The hydrophobic regions face the hydrophobic tails of phospholipids within the membrane bilayer. Membrane proteins can be classified by whether they are embedded (integral) or associated with the cell membrane (peripheral). Most integral proteins are transmembrane proteins, which traverse both phospholipid layers, spanning the entire membrane. Their hydrophilic regions extend from both sides of the membrane, facing cytosol on

 Core: Biology

Microtubules

JoVE 10693

There are three types of cytoskeletal structures in eukaryotic cells—microfilaments, intermediate filaments, and microtubules. With a diameter of about 25 nm, microtubules are the thickest of these fibers. Microtubules carry out a variety of functions that include cell structure and support, transport of organelles, cell motility (movement), and the separation of chromosomes during cell division. Microtubules are hollow tubes whose walls are made up of globular tubulin proteins. Each tubulin molecule is a heterodimer, consisting of a subunit of α-tubulin and a subunit of β-tubulin. The dimers are arranged in linear rows called protofilaments. A microtubule usually consists of 13 protofilaments, arranged side by side, wrapped around the hollow core. Because of this arrangement, microtubules are polar, meaning that they have different ends. The plus end has β-tubulin exposed, and the minus end has α-tubulin exposed. Microtubules can rapidly assemble—grow in length through polymerization of tubulin molecules—and disassemble. The two ends behave differently in this regard. The plus end is typically the fast-growing end or the end where tubulin is added, and the minus end is the slow-growing end or the end where tubulin dissociates—depending on the situation. This process of dynamic instability, where microtu

 Core: Biology

Eukaryotic Compartmentalization

JoVE 10689

One of the distinguishing features of eukaryotic cells is that they contain membrane-bound organelles—such as the nucleus and mitochondria—that carry out particular functions. Since biological membranes are only permeable to a small number of substances, the membrane around an organelle creates a compartment with controlled conditions inside. These microenvironments are often distinct from the environment of the surrounding cytosol and are tailored to the specific functions of the organelle. For example, lysosomes—organelles in animal cells that digest molecules and cellular debris—maintain an environment that is more acidic than the surrounding cytosol, because its enzymes require a lower pH to catalyze reactions. Similarly, pH is regulated within mitochondria, which helps them carry out their function of producing energy. Additionally, some proteins require an oxidative environment for proper folding and processing, but the cytosol is generally reductive. Therefore, these proteins are produced by ribosomes in the endoplasmic reticulum (ER), which maintains the necessary environment. Proteins are often then transported within the cell through membrane-bound vesicles. The genetic material of eukaryotic cells is compartmentalized within the nucleus, which is surrounded by a double membrane called the nuclear envelope. Sma

 Core: Biology

Solvents

JoVE 10670

A solvent is a substance, most often a liquid, that can dissolve other substances. Here, the substance being dissolved is called a solute. When a solvent and a solute combine, they form a solution that, at the molecular level, is a homogenous mixture of both the solvent and the solute. Water is a universal biological solvent. Its polar structure allows it to dissolve many other polar compounds. The ability of water to dissolve is governed by a balance between water molecules binding to each other and binding to the solute. A saturated solution contains the maximum amount of dissolvable solute. For example, salt (NaCl) is readily dissolved in water to create salt water, or saline. It dissolves because salt dissociates into its respective ions sodium (Na+) and chloride (Cl-). Water is polar, so its oxygen atom, being slightly negative, is attracted to the positive sodium ions. Several water molecules can bind to a single sodium ion, creating a sphere of hydration. Likewise, water’s hydrogen atoms are slightly positive and are attracted to the negative chloride ions, again creating a sphere of hydration around the chloride ions. These hydration shells keep the solute particles separated and dispersed, creating a solution. A saturated solution of salt water (at room temperature) contains about 26% sodium chloride. If more sa

 Core: Biology
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