Posts tagged cell membrane
Articles and news from the latest research reports.
(Image caption: Three-dimensional reconstruction of a synapse in the mouse brain. Readily releasable fusionable synaptic vesicles (blue, around 45 millionths of a millimetre in diameter) are docked at the cell membrane. Credit: © MPI f. Experimental Medicine/ Benjamin H. Cooper)
Synapses always on the starting blocks
While neurons rapidly propagate information in their interior via electrical signals, they communicate with each other at special contact points known as the synapses. Chemical messenger substances, the neurotransmitters, are stored in vesicles at the synapses. When a synapse becomes active, some of these vesicles fuse with the cell membrane and release their contents. To ensure that valuable time is not lost, synapses always have some readily releasable vesicles on standby. With the help of high-resolution, three-dimensional electron microscopy, scientists at the Max Planck Institute of Experimental Medicine in Göttingen succeeded in demonstrating that these fusionable vesicles have a very special characteristic: they already have close contact with the cell membrane long before the actual fusion occurs. In addition, the research team also decoded the molecular machinery that facilitates the operation of this docking mechanism.
The fusion of the neurotransmitter vesicles with the cell membrane involves close cooperation between numerous protein components, which monitor each other and ensure that every single ‘participant’ is always in the right place. This is referred to as the fusion machinery and the comparison is an apt one: if a cogwheel in a clock mechanism is broken, the hands do not move. In a similar way, faulty or missing molecules impair synaptic operations.
In research studies carried out some years ago, Nils Brose and his colleague JeongSeop Rhee from the Max Planck Institute of Experimental Medicine in Göttingen already demonstrated that the transmission of information at the synapses in genetically modified mice, in which all known genes of the Munc13 or CAPS proteins had been switched off, is severely defective. Although the neurons of the genetically modified mice do not differ from those of healthy mice when examined under an optical microscope, if Munc13 is missing, the release of neurotransmitters actually grinds to a halt completely. Brose and Rhee’s findings showed that to be able to react immediately to signals at all times, each synapse must keep a small number of ‘readily releasable’ fusionable vesicles on standby.
But how do Munc13 and CAPS convert the vesicles to this kind of fusionable state? To answer this question, the Göttingen-based scientists studied the synaptic contacts in the minutest possible detail. To do this, neurobiologists Cordelia Imig and Ben Cooper, who have been working with Brose and Rhee for many years, used a high-pressure freezing process. This involves the rapid freezing of neurons in the brain tissue under high pressure so that no disruptive ice crystals are formed and the fine structure of the cells is particularly well conserved. The samples obtained in this way were then analysed using electron tomography. Using this method, electron microscope images of a structure are recorded from many different angles, in a similar way to the process used in medical computed tomography. The individual images can then be combined on the computer to give a high-resolution three-dimensional image – of a synapse in this case (see image).
“Our results showed that readily releasable vesicles in healthy synapses touch the cell membrane,” explains Cooper. “However, if Munc13 and CAPS proteins are missing, the vesicles do not reach the active zone and accumulate a few nanometres away from it.” To their astonishment, the researchers also observed that SNARE proteins, which collaborate with Munc13 and CAPS in the nerve endings, are also involved in this docking process. SNARE proteins are found in the cell and vesicle membranes of healthy synapses and control the fusion of the two membranes during neurotransmitter release. When a vesicle approaches the cell membrane, the individual SNARE molecules line up opposite each other like the sides of a zip and pull the membranes close to each other in this way. The vesicles await the starting gun for their fusion in this state – in the starting blocks, so to speak.
The findings of the neurobiologists in Göttingen prove that Munc13, CAPS and SNARE proteins closely align the vesicle and cell membrane in the synapse, long before the signal for fusion is given. This is the only way that the fast and controlled transmission of information at the synapse can be guaranteed, thanks to which we can react specifically to information from our environment. “It had long been clear that synapses have to be extremely fast to carry out all of the many complex brain functions. Our study shows for the first time how this is managed at the molecular level and on the level of the synaptic vesicles,” says Brose. Because almost all of the protein components involved in this process also play a role in neurological and psychiatric diseases, the Göttingen-based scientists believe that their discovery will soon benefit medical research.
(Image caption: Membranes containing monounsaturated (left) and polyunsaturated (right) lipids after adding dynamin and endophilin. In a few seconds membranes rich in polyunsaturated lipids undergo many fissions. Credit: © Mathieu Pinot)
Consuming oils with high polyunsaturated fatty acid content, in particular those containing omega-3s, is beneficial for the health. But the mechanisms underlying this phenomenon are poorly known. Researchers at the Institut de Pharmacologie Moléculaire et Cellulaire (CNRS/Université Nice Sophia Antipolis), the Unité Compartimentation et Dynamique Cellulaires (CNRS/Institut Curie/UPMC), the INSERM and the Université de Poitiers investigated the effect of lipids bearing polyunsaturated chains when they are integrated into cell membranes. Their work shows that the presence of these lipids makes the membranes more malleable and therefore more sensitive to deformation and fission by proteins. These results, published on August 8, 2014 in Science, could help explain the extraordinary efficacy of endocytosis in neuron cells.
Consuming polyunsaturated fatty acids (such as omega-3 fatty acids) is good for the health. The effects range from neuronal differentiation to protection against cerebral ischemia. However the molecular mechanisms underlying these effects are poorly understood, prompting researchers to focus on the role of these fatty acids in cell membrane function.
For a cell to function properly, the membrane must be able to deform and divide into small vesicles. This phenomenon is called endocytosis. Generally, these vesicles allow the cells to encapsulate molecules and transport them. In neurons, these synaptic vesicles will act as a transmission pathway to the synapse for nerve messages. They are formed inside the cell, then they move to its exterior and fuse with its membrane, to transmit the neurotransmitters that they contain. Then they reform in less than a tenth of a second: this is synaptic recycling.
In the work published in Science, the researchers show that cell-or artificial membranes rich in polyunsaturated lipids are much more sensitive to the action of two proteins, dynamin and endophilin, which facilitate membrane deformation and fission.Other measurements in the study and in simulations suggest that these lipids also make the membranes more malleable. By facilitating the deformation and scission necessary for endocytosis, the presence of polyunsaturated lipids could explain rapid synaptic vesicle recycling. The abundance of these lipids in the brain could then represent a major advantage for cognitive function.
This work partially sheds light on the mode of action of omega-3. Considering that the body cannot synthesize them and that they can only be supplied by a suit able diet (rich in oily fish, etc.), it seems important to continue this work to understand the link between the functions performed by these lipids in the neuronal membrane and their health benefits.
Scientists Develop New Process to Create Artificial Cell Membranes
The membranes surrounding and inside cells are involved in every aspect of biological function. They separate the cell’s various metabolic functions, compartmentalize the genetic material, and drive evolution by separating a cell’s biochemical activities. They are also the largest and most complex structures that cells synthesize.
Understanding the myriad biochemical roles of membranes requires the ability to prepare synthetic versions of these complex multi-layered structures, which has been a long-standing challenge.
In a study published this week by Nature Chemistry, scientists at The Scripps Research Institute (TSRI) report a highly programmable and controlled platform for preparing and experimentally probing synthetic cellular structures.
“Layer-by-layer membrane assembly allows us to create synthetic cells with membranes of arbitrary complexity at the molecular and supramolecular scale,” said TSRI Assistant Professor Brian Paegel, who authored the study with Research Associate Sandro Matosevic. “We can now control the molecular composition of the inner and outer layers of a bilayer membrane, and even assemble multi-layered membranes that resemble the envelope of the cell nucleus.”
Starting with a technique commonly used to deposit molecules on a solid surface, Langmuir-Blodgett deposition, the scientists repurposed the approach to work on liquid objects.
The scientists engineered a microfluidic device containing an array of microscopic cups, each trapping a single droplet of water bathed in oil and lipids, the molecules that make up cellular membranes. The trapped droplets are then ready to serve as a foundation for building up a series of lipid layers like coats of paint.
The lipid-coated water droplets are first bathed in water. As the water/oil interface encounters the trapped droplets, a second lipid layer coats the droplets and transforms them into what are known as unilamellar or single-layer vesicles. Bathing the vesicles in oil/lipid deposits a third lipid layer, and followed by a final layer of lipids that is deposited on the trapped drops to yield double-bilayer vesicles.
“The computer-controlled microfluidic circuits we have constructed will allow us to assemble synthetic cells not only from biologically derived lipids, but from any amphiphile and to measure important chemical and physical parameters, such as permeability and stability,” said Paegel.
(Source: scripps.edu)
Researchers develop new pathway to brain for medicine
Stumped for years by a natural filter in the body that allows few substances, including life-saving drugs, to enter the brain through the bloodstream, physicians who treat neurological diseases may soon have a new pathway to the organ via a technique developed by a physicist and an immunologist working together at Florida International University’s Herbert Wertheim College of Medicine.
The FIU researchers developed the technique to deliver and fully release the anti-HIV drug AZTTP into the brain, but their finding has the potential to also help patients who suffer from neurological diseases such as Alzheimer’s, Parkinson’s and epilepsy, as well as cancer.
“Anything where you have trouble getting drugs to the brain and releasing it, this opens so many opportunities,’’ said Madhavan Nair, an FIU professor and chair of the medical school’s immunology department.
In an in vitro laboratory test with HIV-infected cells, Nair and a colleague, Sakhrat Khizroev, a professor of immunology and electrical engineering, attached the antiretroviral drug AZTTP to tiny, magneto-electric nanoparticles. Then, using magnetic energy, they guided the drug across a cell membrane created in the lab to mimic the blood-brain barrier found in the human body.
Once the drug reached its target, researchers triggered its release from the nanoparticle by zapping it with a low-energy electrical current. The drug remained functional and structurally sound after the release, according to the experiment findings.
“We learned to control electrical forces in the brain using magnetics,’’ said Khizroev, who designed, oversaw and supervised the entire project. “We pretty much opened a pathway to the brain.’’
The test findings were published in April in the online peer-reviewed journal, Nature Communications. Researchers believe that using this method will allow physicians to send a higher level of AZTTP — up to 97 percent more — to HIV-infected cells in the brain.
Currently, more than 99 percent of the antiretroviral therapies used to treat HIV, such as AZTTP, are deposited in the liver, lungs and other organs before they reach the brain.
While anti-viral drugs have helped HIV patients live longer by reducing their viral loads, the drugs cannot pass the blood-brain barrier in significant amounts, which allows the virus to lurk unchecked in the brain and can lead to neurological damage, said Dr. Cheryl Holder, a practicing physician and FIU professor who specializes in treating patients with HIV.
“We know that even though the viral load is undetectable in the blood, we don’t know what’s going on in the brain fully,’’ Holder said.
HIV causes constant inflammation, she said, and the virus can pool in areas of the brain where medicine cannot reach, potentially causing damage.
“It’s important to get the drug to the brain,’’ she said, “to help prevent dementia in older patients, and inflammation.’’
But the ability to target drug delivery and release it on demand in the brain has been impossible without opening the skull, Nair and Khizroev said.
Nair, an immunologist who specializes in HIV research, and Khizroev, an electrical engineer and physicist, began collaborating on the project about 18 months ago after winning a National Institutes of Health grant to study the use of magnetic particles.
One of the keys to success was controlling the release of the drug without adversely affecting the brain.
The researchers found their solution in the magneto-electric nanoparticles, which are uniquely suited to deliver and release drugs in the brain, Khizroev said. These nanoparticles can convert magnetic energy into the electrical energy needed to release the drugs without creating heat, which could potentially harm the brain.
The development of a new, less invasive pathway to the brain would open the door to many new medical uses.
Khizroev said he recently returned from a trip to the University of Southern California, where he briefed physicians at the medical school on the technique and its potential for cancer treatment. And Nair said he received a letter recently on behalf of a 91-year-old man suffering from Parkinson’s, asking when the technique might become available for use in people.
That may take a while. With the first phase of testing successfully completed using in vitro experiments, the second will take place at Emory University in Georgia, where researchers will test the technique on monkeys infected with the HIV virus.
If researchers complete the second phase successfully, clinical trials on humans could follow, Nair said. Approval from the Food and Drug Administration would be required before the technique becomes commercially available, he said.
FIU researchers have applied for a patent and would receive royalties, they said, though the university would benefit the most, in part because a successful research project could open opportunities for more grant funding on other topics.
For Khizroev, who had previously done research on quantum computing and information processing, the project has offered a way to put his scientific knowledge to use in a way that could have a direct affect on people’s health.
“I wanted to apply my knowledge of nanoparticles to something important,’’ he said.
(Source: miamiherald.com)
An interdisciplinary team of researchers from the University of Texas Medical Branch at Galveston and the University of Houston has found a new way to influence the vital serotonin signaling system — possibly leading to more effective medications with fewer side effects.
Scientists have linked malfunctions in serotonin signaling to a wide range of health issues, everything from depression and addictions to epilepsy and obesity and eating disorders. Much of their attention has focused on complex proteins called serotonin receptors, which are located in the cell membrane. Each receptor has a so-called “active site” specially suited to bond with a serotonin molecule; when that bond is formed, the receptor changes shape, transmitting a signal to the cell’s interior.
Traditional drug discovery efforts target interactions that take place at such active sites. But a receptor’s behavior can also be changed by additional proteins that bind to the receptor at locations quite distant (in molecular terms) from the active site, in a process called “allosteric regulation” — the mechanism examined by the UTMB-UH team for one specific and highly significant kind of serotonin receptor, designated the 5-HT2C.
“This is a whole new way of thinking about this system, targeting these interactions,” said UTMB professor Kathryn Cunningham, senior author of a paper on the research now online in the Journal of Neuroscience. “Basically, we’ve created a new series of molecules and validated that we can use them to change the way the receptor functions both in vitro and in vivo, through an allosteric effect.”
(Image: thedea.org)