We report the use of chemically modified carbon nanotubes as a substrate for cultured neurons. The morphological features of neurons that directly reflect their potential capability in synaptic transmission are characterized. The chemical properties of carbon nanotubes are systematically varied by attaching different functional groups that confer known characteristics to the substrate. By manipulating the charge carried by functionalized carbon nanotubes we are able to control the outgrowth and branching pattern of neuronal processes.

Astrocytes exhibit excitability based on variations of their intracellular Ca 2ϩ concentrations, which leads to glutamate release, that in turn can signal to adjacent neurons. This glutamate-mediated astrocyte-neuron signaling occurs at physiological intracellular Ca 2ϩ levels in astrocytes and includes modulation of synaptic transmission. The mechanism underlying Ca 2ϩ -dependent glutamate release from astrocytes is most likely exocytosis, because astrocytes express the protein components of the soluble N-ethyl maleimide-sensitive fusion protein attachment protein receptors complex, including synaptobrevin 2, syntaxin, and synaptosome-associated protein of 23 kDa. Although these proteins mediate Ca 2ϩ -dependent glutamate release from astrocytes, it is not well understood whether astrocytes express functional vesicular glutamate transporters (VGLUTs) that are critical for vesicle refilling. Here, we find in cultured and freshly isolated astrocytes the presence of brain-specific Na ϩ -dependent inorganic phosphate cotransporter and differentiation-associated Na ϩ -dependent inorganic phosphate cotransporter that have recently been identified as VGLUTs 1 and 2. Indirect immunocytochemistry showed a punctate pattern of VGLUT immunoreactivity throughout the entire cell body and processes, whereas pharmacological inhibition of VGLUTs abolished mechanically and agonist-evoked Ca 2ϩ -dependent glutamate release from astrocytes. Taken together, these data indicate that VGLUTs play a functional role in exocytotic glutamate release from astrocytes.

We report the synthesis of a single-walled carbon nanotube (SWNT) graft copolymer. This polymer was prepared by the functionalization of SWNTs with polyethyleneimine (PEI). We used this graft copolymer, SWNT-PEI, as a substrate for cultured neurons and found that it promotes neurite outgrowth and branching.

Due to their electrical, chemical, mechanical and thermal properties, carbon nanotubes are one of the most promising materials for the electronics, computer and aerospace industries. Here, we discuss their properties in the context of future applications in biotechnology and biomedicine. The purification and chemical modification of carbon nanotubes with organic, polymeric and biological molecules are discussed. Additionally we review their uses in biosensors, assembly of structures and devices, scanning probe microscopy and as substrates for neuronal growth. We note that additional toxicity studies of carbon nanotubes are necessary so that exposure guidelines and safety regulations can be established in a timely manner. KeywordsCarbon Nanotubes; Structure; Purification; Modification; Biosensors; Assembly of Structures and Devices; Scanning Probe Tips; Nanosurgery; Substrates for Neuronal Growth Although nanomaterials have existed in nature long before mankind was able to identify forms at the nanoscale level, advances in synthetic chemistry have been one of the driving forces in the development of a biological nanotechnology. Nanomaterials have been designed for a variety of biomedical and biotechnological applications, including bone growth, 1 enzyme encapsulation, 2 biosensors 3, 4 and as vesicles for DNA delivery into living cells. 5,6 Whereas nanotechnology may provide novel materials which can result in revolutionary new structures and devices, biotechnology already offers extremely sophisticated tools to precisely position molecules and assemble hierarchal structures and devices. The application of the principles of biology to nanotechnology provides a valuable route for further miniaturization and performance improvement of artificial devices. The feasibility of the bottom-up approach which is based on molecular recognition and self-assembly properties of proteins has already been proved in many inorganic-organic hybrid systems and devices. Nanodevices with biorecognition properties provide tools at a scale, which offers a tremendous opportunity to study biochemical processes and to manipulate living cells at the single molecule level. The synergetic future of nano-and bio-technologies holds great promise for further advancement in tissue engineering, prostheses, genomics, pharmacogenomics, drug delivery, surgery and general medicine.*Author to whom correspondence should be addressed. NIH Public Access NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptEver since Edison discovered that carbon changes its resistance with pressure and a carbon filament glows when an electric current is passed through it, the unique properties of carbon have intrigued scientists. After more than a century of interest, carbon has found its apogee in the fullerenes and carbon nanotubes (CNTs)-arguably the most promising of all nanomaterials. Because of their unique quasi one-dimensional structure and fascinating mechanical and electronic properties, CNTs have captured the attention of physicists...

Astrocytes can respond to a variety of stimuli by elevating their cytoplasmic Ca2+ concentration and can in turn release glutamate to signal adjacent neurons. The majority of this Ca2+ is derived from internal stores while a portion also comes from outside of the cell. Astrocytes use Ca2+ entry through store-operated Ca2+ channels to refill their internal stores. Therefore, we investigated what role this store-operated Ca2+ entry plays in astrocytic Ca2+ responses and subsequent glutamate release. Astrocytes express canonical transient receptor potential (TRPC) channels that have been implicated in mediating store-operated Ca2+ entry. Here, we show that astrocytes in culture and freshly isolated astrocytes from visual cortex express TRPC1, TRPC4, and TRPC5. Indirect immunocytochemistry reveals that these proteins are present throughout the cell; the predominant expression of functionally tested TRPC1, however, is on the plasma membrane. Labeling in freshly isolated astrocytes reveals changes in TRPC expression throughout development. Using an antibody against TRPC1 we were able to block the function of TRPC1 channels and determine their involvement in mechanically and agonist-evoked Ca2+ entry in cultured astrocytes. Blocking TRPC1 was also found to reduce mechanically induced Ca2+-dependent glutamate release. These data indicate that Ca2+ entry through TRPC1 channels contributes to Ca2+ signaling in astrocytes and the consequent glutamate release from these cells.