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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)

Ribosomes

JoVE 10692

Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.

Ribosomes are composed of ribosomal RNA (rRNA) and proteins. Ribosomes are not surrounded by a membrane (i.e., despite their specific cell function, they are not an organelle). In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome production. Within the nucleolus, rRNA is combined with proteins that are imported from the cytoplasm. The assembly produces two subunits of a ribosome—the large and small subunits. These subunits then leave the nucleus through pores in the nuclear envelope. Each one large and small subunit bind to each other once mRNA binds to a site on the small subunit at the start of the translation process. This step forms a functional ribosome. Ribosomes may assemble in the cytosol—called free ribosomes—or while attached to the outside of the nuclear envelope or endoplasmic reticulum—called bound ribosomes. Generally, free ribosomes synthesize proteins used in the cytoplasm, while bound ribosomes synthesize proteins that are inserted into membranes, packaged into org

 Core: Biology

Responses to Salt Stress

JoVE 11120

Salt stress—which can be triggered by high salt concentrations in a plant’s environment—can significantly affect plant growth and crop production by influencing photosynthesis and the absorption of water and nutrients.

Plant cell cytoplasm has a high solute concentration, which causes water to flow from the soil into the plant due to osmosis. However, excess salt in the surrounding soil increases the soil solute concentration, reducing the plant’s ability to take up water. High levels of sodium are toxic to plants, so increasing their sodium content to compensate is not a viable option. However, many plants can respond to moderate salt stress by increasing internal levels of solutes that are well-tolerated at high concentrations—like proline and glycine. The resulting increased solute concentration within the cell cytoplasm allows the roots to increase water uptake from the soil without taking in toxic levels of sodium. Sodium is not essential for most plants, and excess sodium affects the absorption of essential nutrients. For example, the uptake of potassium—which regulates photosynthesis, protein synthesis, and other essential plant functions—is impeded by sodium in highly saline conditions. Calcium can ameliorate some effects of salt stress by facilitating potassium uptake through the regulation of ion

 Core: Biology

Types of Hormones

JoVE 10988

Hormones can be classified into three main types based on their chemical structures: steroids, peptides, and amines. Their actions are mediated by the specific receptors they bind to on target cells.

Steroid hormones are derived from cholesterol and are lipophilic in nature. This allows them to readily traverse the lipid-rich cell membrane to bind to their intracellular receptors in the cytoplasm or nucleus. Once bound, the cytoplasmic hormone-receptor complex translocates to the nucleus. Here, it binds to regulatory sequences on the DNA to alter gene expression. Peptide hormones are made up of chains of amino acids and are hydrophilic. Hence, they are unable to diffuse across the cell membrane. Instead, they bind to extracellular receptors present on the surface of target cells. Such binding triggers a series of signaling reactions within the cell to ultimately carry out the specific functions of the hormone. Amine hormones are derived from a single amino acid, either tyrosine or tryptophan. This class of hormones is unique because they share their mechanism of action with both steroid as well as peptide hormones. For example, although epinephrine and thyroxine are both derived from the amino acid tyrosine, they mediate their effects through diverse mechanisms. Epinephrine binds to G-protein coupled receptors present on the surface of the plasma

 Core: Biology

MicroRNAs

JoVE 10801

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends have been methylated to prevent degradation, it is exported from the nucleus into the cytoplasm. In the cytoplasm, another endonuclease enzyme, called Dicer, cuts the pre-miRNA into a 21-24 nucleotide-long miRNA duplex. Then, Dicer cleaves one strand of the duplex, releasing a single strand of mature miRNA. The mature miRNA is loaded into a protein complex called RNA-induced silencing complex (RISC), which the miRNA then guides to the complementary region of its target mRNA. The extent of complementary base-pairing between miRNA and 3’ untranslated region of target mRNA determines the gene silencing mechanism. Extensive or near-perfect complementarity causes degradation of mRNA, whereas limited base-pairing inhibits translation. While silencing via mRNA degradation is irreversible, translation inhibition is reversible since stable mRNA can

 Core: Biology

Translation

JoVE 10795

Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA) and other proteins are involved in the production of the chain of amino acids—the polypeptide. Proteins are called the “building blocks” of life because they make up the vast majority of all organisms—from muscle fibers to hairs on your head to components of your immune system—and the blueprint for each and every one of those proteins is encoded by the genes found in the DNA of every cell. The central dogma in biology dictates that genetic information is converted into functional proteins by the processes of transcription and translation. Eukaryotes have a membrane-bound nucleus where mRNA is transcribed from DNA. After transcription, mRNA is shuttled out of the nucleus to be translated into a chain of amino acids—a polypeptide—and eventually, a functional protein. This can take place in the cytoplasm or in the rough endoplasmic reticulum, where the polypeptides are further modified. By contrast, prokaryotes lack a nuclear compartment, so translation in prokaryotes takes place in the cytoplasm,

 Core: Biology

Phagocytosis

JoVE 10710

Cells pull particles inward and engulf them in spherical vesicles in an energy-requiring process called endocytosis. Phagocytosis (“cellular eating”) is one of three major types of endocytosis. Cells use phagocytosis to take in large objects—such as other cells (or their debris), bacteria, and even viruses.

The objective of phagocytosis is often destruction. Cells use phagocytosis tonate unwelcome visitors, like pathogens (e.g., viruses and bacteria). It is perhaps unsurprising, that many immune system cells, including neutrophils, macrophages, and monocytes, leverage phagocytosis to destroy pathogens or infected host cells. In addition to immune system cells, amoebae, algae, and other single-celled organisms use phagocytosis to eat. Phagocytosis begins when a particle (e.g., virus) contacts the engulfing cell, called a phagocyte. Sometimes, this is a chance encounter. Other times, the phagocyte follows a chemical signal to the particle, in a process called chemotaxis. The phagocyte eventually binds to the particle or cell via surface receptors. Different types of phagocytes use distinct receptors for phagocytosis. These receptors may be general, responding to a variety of stimuli, or specific. The phagocyte begins to surround and engulf the particle bound to its surface by extending regions of its cytoplasm, called pseudopods, around the particle.

 Core: Biology

Diffusion and Osmosis- Concept

JoVE 10622

Cell Membranes and Diffusion

In order to function, cells are required to move materials in and out of their cytoplasm via their cell membranes. These membranes are semipermeable, meaning that certain molecules are allowed to pass through, but not others. This movement of molecules is mediated by the phospholipid bilayer and its embedded proteins, some of which act as transport channels…

 Lab Bio

Zebrafish Microinjection Techniques

JoVE 5130

One of the major advantages to working with zebrafish (Danio rerio) is that their genetics can be easily manipulated by microinjection of early stage embryos. Using this technique, solutions containing genetic material or knockdown constructs are delivered into the blastomeres: the embryonic cells sitting atop the yolk of the newly fertilized egg. Delivery into the cytoplasm is…

 Biology II

An Introduction to Transfection

JoVE 5068

Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for…

 Basic Methods in Cellular and Molecular Biology
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