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DOE OSTI2020

Transcriptomic and proteomic signatures of stemness and differentiation in the colon crypt

The intestinal epithelium is replaced weekly by non-quiescent stem cells with kinetics that rely on a rapid loss of stemness and choice for secretory or absorptive lineage differentiation. To determine how the cellular transcriptome and proteome changes during these transitions, we developed a new cell sorting method to purify stem cells, secretory and absorptive progenitor cells, and mature, differentiated cells. Transcriptome analyses revealed that as stem cells transition to the progenitor stage, alternative mRNA splicing and polyadenylation dominate changes in the transcriptome. In contrast, as progenitors differentiate into mature cell types, alterations in gene expression and mRNA levels drive the changes. RNA processing targets mRNAs encoding regulators of cell cycle, RNA regulators, cell adhesion, SUMOylation, and Wnt and Notch signaling. Additionally, global proteome profiling of sorted cell populations detected >2,800 proteins and revealed RNA:protein patterns of abundance and correlation. Paired together, these data highlight new potentials for autocrine and feedback regulation and provide new insights into cell state transitions in the crypt.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

Sorted-cell proteomics reveals an AT1-associated epithelial cornification phenotype and suggests endothelial redox imbalance in human bronchopulmonary dysplasia

Bronchopulmonary dysplasia (BPD) is a neonatal lung disease characterized by inflammation and scarring leading to long-term tissue damage. Previous whole tissue proteomics identified BPD-specific proteome changes and cell type shifts. Little is known about the proteome-level changes within specific cell populations in disease. Here, we sorted epithelial (EPI) and endothelial (ENDO) cell populations based on their differential surface markers from normal and BPD human lungs. Using a low-input compatible sample preparation method (MicroPOT), proteins were extracted and digested into peptides and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteome analysis. Of the 4,970 proteins detected, 293 were modulated in abundance or detection in the EPI population and 422 were modulated in ENDO cells. Modulation of proteins associated with actin-cytoskeletal function, such as SCEL, LMO7, and TBA1B was observed in the BPD EPIs. Using confocal imaging and analysis, we validated the presence of aberrant multilayer-like structures comprising SCEL and LMO7, known to be associated with epidermal cornification, in the human BPD lung. This is the first report of the accumulation of cornification-associated proteins in BPD. Their localization in the alveolar parenchyma, primarily associated with alveolar type 1 (AT1) cells, suggests a role in the BPD postinjury response. In the ENDOs, redox balance and mitochondrial function pathways were modulated. Alternative mRNA splicing and cell proliferative functions were elevated in both populations, suggesting potential dysregulation of cell progenitor fate. This study characterized the proteome of epithelial and endothelial cells from the BPD lung for the first time, identifying population-specific changes in BPD pathogenesis.

BPD↗

NASA NTRS2001

Extracellular calcium sensing and extracellular calcium signaling

The cloning of a G protein-coupled extracellular Ca(2+) (Ca(o)(2+))-sensing receptor (CaR) has elucidated the molecular basis for many of the previously recognized effects of Ca(o)(2+) on tissues that maintain systemic Ca(o)(2+) homeostasis, especially parathyroid chief cells and several cells in the kidney. The availability of the cloned CaR enabled the development of DNA and antibody probes for identifying the CaR's mRNA and protein, respectively, within these and other tissues. It also permitted the identification of human diseases resulting from inactivating or activating mutations of the CaR gene and the subsequent generation of mice with targeted disruption of the CaR gene. The characteristic alterations in parathyroid and renal function in these patients and in the mice with "knockout" of the CaR gene have provided valuable information on the CaR's physiological roles in these tissues participating in mineral ion homeostasis. Nevertheless, relatively little is known about how the CaR regulates other tissues involved in systemic Ca(o)(2+) homeostasis, particularly bone and intestine. Moreover, there is evidence that additional Ca(o)(2+) sensors may exist in bone cells that mediate some or even all of the known effects of Ca(o)(2+) on these cells. Even more remains to be learned about the CaR's function in the rapidly growing list of cells that express it but are uninvolved in systemic Ca(o)(2+) metabolism. Available data suggest that the receptor serves numerous roles outside of systemic mineral ion homeostasis, ranging from the regulation of hormonal secretion and the activities of various ion channels to the longer term control of gene expression, programmed cell death (apoptosis), and cellular proliferation. In some cases, the CaR on these "nonhomeostatic" cells responds to local changes in Ca(o)(2+) taking place within compartments of the extracellular fluid (ECF) that communicate with the outside environment (e.g., the gastrointestinal tract). In others, localized changes in Ca(o)(2+) within the ECF can originate from several mechanisms, including fluxes of calcium ions into or out of cellular or extracellular stores or across epithelium that absorb or secrete Ca(2+). In any event, the CaR and other receptors/sensors for Ca(o)(2+) and probably for other extracellular ions represent versatile regulators of numerous cellular functions and may serve as important therapeutic targets.

Review↗

DOE OSTI2021

An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein

Protein splicing is a post-translational process by which an intein catalyzes its own excision from flanking polypeptides, or exteins, concomitant with extein ligation. Many inteins have nested homing endonuclease domains that facilitate their propagation into intein-less alleles, whereas other inteins lack the homing endonuclease (HEN) and are called mini-inteins. The mini-intein that interrupts the DNA PolII of Pyrococcus horikoshii has a linker region in place of the HEN domain that is shorter than the linker in a closely related intein from Pyrococcus abyssi. The P. horikoshii PolII intein requires a higher temperature for catalytic activity and is more stable to digestion by the thermostable protease thermolysin, suggesting that it is more rigid than the P. abyssi intein. We solved a crystal structure of the intein precursor that revealed a domain-swapped dimer. Inteins found as domain swapped dimers have been shown to promote intein-mediated protein alternative splicing, but the solved P. horikoshii PolII intein structure has an active site unlikely to be catalytically competent.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2021

A Deep Exon Cryptic Splice Site Promotes Aberrant Intron Retention in a Von Willebrand Disease Patient

A translationally silent single nucleotide mutation in exon 44 (E44) of the von Willebrand factor (VWF) gene is associated with inefficient removal of intron 44 in a von Willebrand disease (VWD) patient. This intron retention (IR) event was previously attributed to reordered E44 secondary structure that sequesters the normal splice donor site. We propose an alternative mechanism: the mutation introduces a cryptic splice donor site that interferes with the function of the annotated site to favor IR. We evaluated both models using minigene splicing reporters engineered to vary in secondary structure and/or cryptic splice site content. Analysis of splicing efficiency in transfected K562 cells suggested that the mutation-generated cryptic splice site in E44 was sufficient to induce substantial IR. Mutations predicted to vary secondary structure at the annotated site also had modest effects on IR and shifted the balance of residual splicing between the cryptic site and annotated site, supporting competition among the sites. Further studies demonstrated that introduction of cryptic splice donor motifs at other positions in E44 did not promote IR, indicating that interference with the annotated site is context dependent. We conclude that mutant deep exon splice sites can interfere with proper splicing by inducing IR.

60 APPLIED LIFE SCIENCES↗

NASA NTRS1991

Stiff, Strong Splice For A Composite Sandwich Structure

New type of splice for composite sandwich structure reduces peak shear stress in structure. Layers of alternating fiber orientation interposed between thin ears in adhesive joint. Developed for structural joint in spar of helicopter rotor blade, increases precision of control over thickness of adhesive at joint. Joint easy to make, requires no additional pieces, and adds little weight.

Schmaling, D.↗

DOE OSTI2024

Long- and short-read sequencing methods discover distinct circular RNA pools in Lotus japonicus

Circular RNAs (circRNAs) are covalently closed single-stranded RNAs, generated through a back-splicing process that links a downstream 5′ site to an upstream 3′ end. The only distinction in the sequence between circRNA and their linear cognate RNA is the back splice junction. Their low abundance and sequence similarity with their linear origin RNA have made the discovery and identification of circRNA challenging. We have identified almost 6000 novel circRNAs from Lotus japonicus leaf tissue using different enrichment, amplification, and sequencing methods as well as alternative bioinformatics pipelines. The different methodologies identified different pools of circRNA with little overlap. We validated circRNA identified by the different methods using reverse transcription polymerase chain reaction and characterized sequence variations using nanopore sequencing. We compared validated circRNA identified in L. japonicus to other plant species and showed conservation of high-confidence circRNA-expressing genes. This is the first identification of L. japonicus circRNA and provides a resource for further characterization of their function in gene regulation. CircRNAs identified in this study originated from genes involved in all biological functions of eukaryotic cells. The comparison of methodologies and technologies to sequence, identify, analyze, and validate circRNA from plant tissues will enable further research to characterize the function and biogenesis of circRNA in L. japonicus.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2026

Expanding the genetic toolkit: adenine and cytosine base editors for gene disruption in Aspergillus niger

Despite revolutionizing fungal genetic engineering, conventional CRISPR/Cas9-mediated knockouts rely on DNA double-strand breaks (DSBs), which can cause unwanted insertions and deletions, chromosomal abnormalities, and cytotoxicity. Base editors such as adenine base editors (ABEs), which convert A‧T to G‧C, and cytosine base editors (CBEs), which convert C‧G to T‧A, offer a safer alternative by enabling predictable, target-specific single-nucleotide changes without introducing DSBs. To overcome the limitations of traditional genome editing in filamentous fungi, we developed efficient base-editing systems in Aspergillus niger . For the first time, we constructed an ABE in A. niger , achieving up to 80% editing efficiency and inducing predictable A-to-G mutations at the intended intron sites, disrupting gene function through mRNA mis-splicing. We also developed a highly efficient CBE system, capable of introducing premature stop codons with 50–100% efficiency. To broaden the editing scope, we implemented a Cas9-NG variant recognizing a relaxed PAM sequence requiring only a single guanine (G), enabling editing at start codons and splice sites. Leveraging this expanded scope, we established gene disruption approaches by targeting start codons via ABE-mediated A-to-G conversions (ATG-to-GTG and ATG-to-ACG) and CBE-mediated C-to-T conversion (ATG-to-ATA). Additionally, our base-editing systems enable multiplex gRNA delivery and marker-free editing of multiple genes. Collectively, the scope-expanding strategies increase the number of genes targetable for disruption by base-editing in A. niger by 26.3% and enable near-complete coverage of 96% of the coding genes. Overall, this work demonstrates the potential of ABE and CBE systems as versatile, efficient, and safer alternatives to DSBs-based gene disruption in filamentous fungi.

Aspergillus↗

DOE OSTI2026

Measurement of Transverse Resistance for Stacks of Non-insulated REBCO Tapes

Transverse resistance among adjacent conductors is a key parameter to calculate AC losses in superconducting cables with non-insulated conductors. The transverse pressure insert (TPI) previously developed and used at Fermilab was modified to measure transverse resistance of stacks made of non-insulated REBCO tapes as a function of transverse pressure at liquid nitrogen temperature. Pressure up to 322 MPa was applied with a hydraulic cylinder. A small current up to 10 A was flown in the stack sample through REBCO tape leads spliced above and below the stack. The latter had a bending radius larger than 6 mm to prevent the critical current degradation in REBCO leads. The voltage was measured just outside the compressed area. Stack samples included stacks of bare 4-mm wide REBCO tapes, as well as REBCO tapes alternated with 4-mm wide stainless-steel ribbons of 40 μm and 30 μm of thickness. We herein present the results of these transverse resistance measurements. Results show a stronger dependence on pressure for smaller pressure vs. larger ones. Also, of the two components of contact resistance, i.e. contact and bulk, the latter was negligible.

Tamagnini, E. [Turin Polytechnic]↗

NASA NTRS1980

Design studies of Laminar Flow Control (LFC) wing concepts using superplastics forming and diffusion bonding (SPF/DB)

Alternate concepts and design approaches were developed for suction panels and techniques were defined for integrating these panel designs into a complete LFC 200R wing. The design concepts and approaches were analyzed to assure that they would meet the strength, stability, and internal volume requirements. Cost and weight comparisions of the concepts were also made. Problems of integrating the concepts into a complete aircraft system were addressed. Methods for making splices both chordwise and spanwise, fuel light joints, and internal duct installations were developed. Manufacturing problems such as slot aligment, tapered slot spacing, production methods, and repair techniques were addressed. An assessment of the program was used to developed recommendations for additional research in the development of SPF/DB for LFC structure.

Wilson, V. E.↗

NASA NTRS1999

A complex ligase ribozyme evolved in vitro from a group I ribozyme domain

Like most proteins, complex RNA molecules often are modular objects made up of distinct structural and functional domains. The component domains of a protein can associate in alternative combinations to form molecules with different functions. These observations raise the possibility that complex RNAs also can be assembled from preexisting structural and functional domains. To test this hypothesis, an in vitro evolution procedure was used to isolate a previously undescribed class of complex ligase ribozymes, starting from a pool of 10(16) different RNA molecules that contained a constant region derived from a large structural domain that occurs within self-splicing group I ribozymes. Attached to this constant region were three hypervariable regions, totaling 85 nucleotides, that gave rise to the catalytic motif within the evolved catalysts. The ligase ribozymes catalyze formation of a 3',5'-phosphodiester linkage between adjacent template-bound oligonucleotides, one bearing a 3' hydroxyl and the other a 5' triphosphate. Ligation occurs in the context of a Watson-Crick duplex, with a catalytic rate of 0.26 min(-1) under optimal conditions. The constant region is essential for catalytic activity and appears to retain the tertiary structure of the group I ribozyme. This work demonstrates that complex RNA molecules, like their protein counterparts, can share common structural domains while exhibiting distinct catalytic functions.

NASA Discipline Exobiology↗

NASA NTRS2002

Commercial-Off-The-Shelf Microelectromechanical Systems (MEMS) Flow-Measurement Probes Fabricated And Assembled

As an alternative to conventional tubing instrumentation for measuring airflow, designers and technicians at the NASA Glenn Research Center have been fabricating packaging components and assembling a set of unique probes using commercial-off-the-shelf microelectromechanical systems (MEMS) integrated circuits (computer chips). Using MEMS as an alternative has some compelling advantages over standard measurement devices. Sensor technologies have matured through high-production usage in industries such as automotive and aircraft manufacturers. Currently, MEMS are the choice in applications such as tire pressure monitors, altimeters, pneumatic controls, cable leak detectors, and consumer appliances. Conventional instrumentation uses tubing buried in the model aerodynamic surfaces or wind tunnel walls. The measurements are made when pressure is introduced at the tube opening. The pressure then must travel the tubing for lengths ranging from 20 to hundreds of feet before reaching an electronic signal conditioner. This condition causes a considerable amount of damping and requires measurements to be made only after the test rig has reached steady-state operation. The electronic MEMS pressure sensor is able to take readings continuously under dynamic states in nearly real time. The use of stainless steel tubing for pressure measurements requires many tubes to be cleaned, cut to length, carefully installed, and delicately deburred and spliced for use. A cluster of a few hundred 1/16-in.- (0.0625-in.-) diameter tubes (not uncommon in research testing facilities) can be several inches in diameter and may weigh enough to require two men to handle. Replacing hard tubing with electronic chips can eliminate much of the bulk. Each sensor would fit on the tip of the 1/16-in. tubing with room to spare. The P592 piezoresistive silicon pressure sensor (Lucas NovaSensor, Fremont, CA) was chosen for this project because of its cost, availability, and tolerance to extreme ambient conditions. The chip is 1 mm square by 0.6 mm thick (0.039 by 0.023 in.) with 0.12-mm (0.005-in.) wire connection tabs. Three MEMS chips were used to build the first type of flow-angularity probe. This MEMS probe will be demonstrated as an alternative to a standard tube type "Cobra Probe" now used routinely in wind tunnel and aeronautical hardware applications. The response time and accuracy would allow the probe to be translated on an actuator across a flow field, yielding precision dynamic measurements not possible with conventional instrumentation. The low profile, the minimal power requirement, the rugged construction, and the moderate cost all contribute to making MEMS sensors the enticing choice instrument in future research measurement needs. The MEMS probe efforts are a continuation of work initiated by Brian Willis, without whose foresight and efforts this project would never have been realized. This task was funded through cooperation with the NASA Electronic Parts and Packaging (NEPP) program at the Jet Propulsion Laboratory.

Redding, Chip↗

NASA NTRS1991

Permanent wire splicing by an explosive joining process

The invention is an apparatus and method for wire splicing using an explosive joining process. The apparatus consists of a prebent, U-shaped strap of metal that slides over prepositioned wires. A standoff means separates the wires from the strap before joining. An adhesive means holds two ribbon explosives in position centered over the U-shaped strap. A detonating means connects to the ribbon explosives. The process involves spreading strands of each wire to be joined into a flat plane. The process then requires alternating each strand in alignment to form a mesh-like arrangement with an overlapped area. The strap slides over the strands of the wires, and the standoff means is positioned between the two surfaces. The detonating means then initiates the ribbon explosives that drive the strap to accomplish a high velocity, angular collision between the mating surfaces. This collision creates surface melts and collision bonding results in electron sharing linkups.

Bement, Laurence J.↗

NASA NTRS1997

Explosive Spot Joining of Metals

The invention is an apparatus and method for wire splicing using an explosive joining process. The apparatus consists of a prebend, U-shaped strap of metal that slides over prepositioned wires. A standoff means separates the wires from the strap before joining. An adhesive means holds two ribbon explosives in position centered over the U-shaped strap. A detonating means connects to the ribbon explosives. The process involves spreading strands of each wire to be joined into a flat plane. The process then requires alternating each strand in alignment to form a mesh-like arrangement with an overlapped area. The strap slides over the strands of the wires. and the standoff means is positioned between the two surfaces. The detonating means then initiates the ribbon explosives that drive the strap to accomplish a high velocity. angular collision between the mating surfaces. This collision creates surface melts and collision bonding resulting in electron-sharing linkups.

Bement, Laurence J.↗

NASA NTRS2008

Improved Gas Filling and Sealing of an HC-PCF

An improved packaging approach has been devised for filling a hollow-core photonic-crystal fiber (HC-PCF) with a gas, sealing the HC-PCF to retain the gas, and providing for optical connections and, optionally, a plumbing fitting for changing or augmenting the gas filling. Gas-filled HC-PCFs can be many meters long and have been found to be attractive as relatively compact, lightweight, rugged alternatives to conventional gas-filled glass cells for use as molecular-resonance frequency references for stabilization of lasers in some optical-metrology, lidar, optical-communication, and other advanced applications. Prior approaches to gas filling and sealing of HC-PCFs have involved, variously, omission of any attempt to connectorize the PCF, connectorization inside a vacuum chamber (an awkward and expensive process), or temporary exposure of one end of an HC-PCF to the atmosphere, potentially resulting in contamination of the gas filling. Prior approaches have also involved, variously, fusion splicing of HC-PCFs with other optical fibers or other termination techniques that give rise to Fresnel reflections of about 4 percent, which results in output intensity noise.

Poberezhskiy, Ilya↗

NASA NTRS2004

COTS MEMS Flow-Measurement Probes

As an alternative to conventional tubing instrumentation for measuring airflow, designers and technicians at Glenn Research Center have been fabricating packaging components and assembling a set of unique probes that contain commercial off-the-shelf (COTS) microelectromechanical systems (MEMS) sensor chips. MEMS sensor chips offer some compelling advantages over standard macroscopic measurement devices. MEMS sensor technology has matured through mass production and use in the automotive and aircraft industries. At present, MEMS are the devices of choice for sensors in such applications as tire-pressure monitors, altimeters, pneumatic controls, cable leak detectors, and consumer appliances. Compactness, minimality of power demand, rugged construction, and moderate cost all contribute to making MEMS sensors attractive for instrumentation for future research. Conventional macroscopic flow-measurement instrumentation includes tubes buried beneath the aerodynamic surfaces of wind-tunnel models or in wind-tunnel walls. Pressure is introduced at the opening of each such tube. The pressure must then travel along the tube before reaching a transducer that generates an electronic signal. The lengths of such tubes typically range from 20 ft (approx.= 6 m) to hundreds of feet (of the order of 100 m). The propagation of pressure signals in the tubes damps the signals considerably and makes it necessary to delay measurements until after test rigs have reached steady-state operation. In contrast, a MEMS pressure sensor that generates electronic output can take readings continuously under dynamic conditions in nearly real time. In order to use stainless-steel tubing for pressure measurements, it is necessary to clean many tubes, cut them to length, carefully install them, delicately deburr them, and splice them. A cluster of a few hundred 1/16-in.- (approx.=1.6-mm-) diameter tubes (such clusters are common in research testing facilities) can be several inches (of the order of 10 cm) in diameter and could weigh enough that two technicians are needed to handle it. Replacing hard tubing with electronic chips can eliminate much of the bulk. Each sensor would fit on the tip of a 1/16-in. tube with room to spare. The Lucas NovaSensor P592 piezoresistive silicon pressure sensor was chosen for this project because of its cost, availability, and tolerance to extreme ambient conditions. The sensor chip is 1 mm square by 0.6 mm thick (about 0.039 by 0.039 by 0.024 in.) and includes 0.12-mm (approx.=0.005-in.) wire connection tabs. The figure shows a flow-angularity probe that was built by use of three such MEMS chips. It is planned to demonstrate this MEMS probe as an alternative to a standard tube-type "Cobra" probe now used routinely in wind tunnels and aeronautical hardware. This MEMS probe could be translated across a flow field by use of a suitable actuator, so that its accuracy and the shortness of its response time could be exploited to obtain precise dynamic measurements of a sort that cannot be made by use of conventional tubing-based instrumentation.

Redding, Chip↗

DOE OSTI2024

Distributed Magnetic Field and Temperature Monitoring for Superconducting Radio Frequency Cavities

The overall objective of the proposed Phase I program was to design, construct and demonstrate a fiber optic sensing system capable of providing temperature and magnetic field measurements with an enhanced spatial resolution that can be implemented over a large surface area (cryomodules) to survey superconducting radio frequency cavities and magnets. A magnetic field sensor capable of detecting fluxes on the order of 1 μT is required to detect the distribution of trapped flux on the cavity surface. A unique distributed magnetic field sensor was successfully designed and constructed to demonstrate the detection of magnetic fluxes less than 500 nT. The sensor leveraged the ultra-high sensitivity of Sentek’s picoDAS to measure the magnetostriction induced vibrations in a commercially available Metglas 2605 SC ribbon that was in physical contact with sensing fiber. Static magnetic fields were detected by applying an alternating current a copper wire proximate to the Metglas 2605SC ribbon to create an AC bias magnetic field. In an alternative approach, an AC bias magnetic field was applied to a special magnetic field sensing fiber with Metglas 2605SC cladding successfully detect a magnetic field flux of a 3 μT. Exhaustive testing was performed to characterize the dependency of sensor response on the direction of the applied magnetic field. Although the special sensing fiber based magnetic field sensor did not exhibit an observable dependence on the direction of the magnetic field, the Metglas 2605SC ribbon sensor exhibited a clear directional dependence. A wide variety of polymer materials were evaluated to enhance the temperature response of an FBG based sensor at cryogenic temperatures. The processing and performance challenges provided the motivation to develop a new simple cryogenic temperature sensor that uses a commercially available fiber optic splice protector. The EVA hot melt tube that becomes adhered to the optical fiber and the polyolefin outer tube that shrinks upon heating in the fusion splicer heater provide the high thermal expansion coefficient necessary to impart a significant strain on the FBG when exposed to cryogenic temperatures. The temperature sensitivity (Δ𝜆𝐵𝑟𝑎𝑔𝑔~ 62 𝑝𝑝𝑚/℃) of the FBG-based sensor was on par with the best reported to date. The simple design, use of readily available cost-effective materials, and well-established processing techniques lends this approach to the creation of hundreds to thousands of temperature sensors on one single optical fiber length. The inherently small form factor also allows for co-location with the distributed magnetic field sensor. In preparation for field testing of the prototype sensing system at the Jefferson Labs in potential Phase II program, several different cable designs were evaluated to package the sensors. The preliminary successful demonstration of fully functional sensing cables provides the foundation for subsequent development efforts to advance the Technology Readiness Level of the technology. The technical feasibility of the proposed approach was successfully demonstrated in this Phase I effort.

43 PARTICLE ACCELERATORS↗

NASA NTRS2001

Developmental regulation of N-methyl-D-aspartate- and kainate-type glutamate receptor expression in the rat spinal cord

Spinal motor neurons undergo experience-dependent development during a critical period in early postnatal life. It has been suggested that the repertoire of glutamate receptor subunits differs between young and mature motor neurons and contributes to this activity-dependent development. In the present study we examined the expression patterns of N-methyl-D-aspartate- and kainate-type glutamate receptor subunits during the postnatal maturation of the spinal cord. Young motor neurons express much higher levels of the N-methyl-D-aspartate receptor subunit NR1 than do adult motor neurons. Although there are eight potential splice variants of NR1, only a subgroup is expressed by motor neurons. With respect to NR2 receptor subunits, young motor neurons express NR2A and C, while adult motor neurons express only NR2A. Young motor neurons express kainate receptor subunits GluR5, 6 and KA2 but we are unable to detect these or any other kainate receptor subunits in the adult spinal cord. Other spinal cord regions display a distinct pattern of developmental regulation of N-methyl-D-aspartate and kainate receptor subunit expression in comparison to motor neurons. Our findings indicate a precise spatio-temporal regulation of individual subunit expression in the developing spinal cord. Specific combinations of subunits in developing neurons influence their excitable properties and could participate in the emergence of adult neuronal form and function.

NASA Discipline Cell Biology↗