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> Physiology > Organism Function > Neuronal Outgrowth

Neuronal Outgrowth

Neuronal Outgrowth refers to the process by which neurons extend projections, known as axons and dendrites, to form connections with other cells.
This critical developmental process is essential for the formation and refinement of neural circuits.
Researchers studying Neuronal Outgrowth may investigate factors that promote or inhiibt this growth, such as signaling molecules, extracellular matrix components, and genetic regulators.
Understanding the mechanisms underlying Neuronal Outgrowth can provide insights into nervous system development, regeneration, and neurological disorders.

Most cited protocols related to «Neuronal Outgrowth»

1
Generating Human iPSC-Derived Neurons for Neuroscience Research
Cited 61 times
Embryoid bodies were generated from hiPSCs and then transferred to nonadherent plates (Corning). Colonies were maintained in suspension in N2 media (DMEM/F12 (Invitrogen), 1x N2 (Invitrogen)) for 7 days and then plated onto polyornithine (PORN)/Laminin-coated plates. Visible rosettes formed within 1 week and were manually dissected and cultured in NPC media (DMEM/F12, 1x N2, 1x B27-RA (Invitrogen), 1 µg/ml Laminin (Invitrogen) and 20 ng/ml FGF2 (Invitrogen). NPCs are maintained at high density, grown on PORN/Laminin-coated plates in NPC media and split approximately 1:4 every week with Accutase (Millipore). For neural differentiations, NPCs were dissociated with Accutase and plated at low density in neural differentiation media (DMEM/F12-Glutamax, 1x N2, 1x B27-RA, 20 ng/ml BDNF (Peprotech), 20 ng/ml GDNF (Peprotech), 1 mm dibutyrl-cyclicAMP (Sigma), 200 nM ascorbic acid (Sigma) onto PORN/Laminin-coated plates.
Assays for neuronal connectivity, neurite outgrowth, synaptic protein expression, synaptic density, electrophysiology, spontaneous calcium transient imaging and gene expression were used to compare control and SCZD hiPSC neurons.
Additional methods are found in S.I.
Brennand K., Simone A., Jou J., Gelboin-Burkhart C., Tran N., Sangar S., Li Y., Mu Y., Chen G., Yu D., McCarthy S., Sebat J, & Gage F.H. (2011). Modeling schizophrenia using hiPSC neurons. Nature, 473(7346), 221-225.
Publication 2011
accutase Ascorbic Acid Biological Assay Calcium Culture Media Embryoid Bodies Fibroblast Growth Factor 2 GDNF protein, human Gene Expression Human Induced Pluripotent Stem Cells Laminin Nervousness Neuronal Outgrowth Neurons polyornithine Proteins Schizophrenia Transients
2
PC12 Cell Culture and Differentiation
Cited 26 times
Because of the clonal instability of the PC12 cell line (Fujita et al. 1989 (link)), the experiments were performed on cells that had undergone fewer than five passages, and all studies were repeated several times with different batches of cells. As described previously (Crumpton et al. 2000a (link); Qiao et al. 2003 (link); Song et al. 1998 (link)), PC12 cells (1721-CRL; American Type Culture Collection, Manassas, VA) were seeded onto 100-mm poly-d-lysine-coated plates in RPMI-1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% inactivated horse serum (Sigma Chemical Co., St. Louis, MO), 5% fetal bovine serum (Sigma Chemical Co.), and 50 μg/mL penicillin streptomycin (Invitrogen). Cells were incubated with 7.5% CO2 at 37°C, and the medium was changed every 2 days. For studies in the undifferentiated state, cells were seeded at varying densities so that, regardless of the total time of incubation, the cells would reach a final confluence of 60–70%. Twenty-four hours after seeding, the medium was changed to include the various test substances: chlorpyrifos (Chem Service, West Chester, PA), diazinon (Chem Service), parathion (Chem Service), physostigmine (Sigma Chemical Co.), dieldrin (Chem Service), or NiCl2 (Sigma Chemical Co.). Because of their poor water solubility, the pesticides were dissolved in dimethyl sulfoxide (Sigma Chemical Co.), achieving a final concentration of 0.1% in the culture medium; accordingly, all cultures included this vehicle, which had no effect on the PC12 cells (Qiao et al. 2001 (link), 2003 (link); Song et al. 1998 (link)).
For studies in differentiating cells, 3 × 106 cells were seeded; 24 hr later, the medium was changed to include 50 ng/mL 2.5 S murine NGF (Invitrogen), and each culture was examined under a microscope to verify the subsequent outgrowth of neurites. The test agents were added concurrently with the start of NGF treatment.
Slotkin T.A., MacKillop E.A., Ryde I.T., Tate C.A, & Seidler F.J. (2006). Screening for Developmental Neurotoxicity Using PC12 Cells: Comparisons of Organophosphates with a Carbamate, an Organochlorine, and Divalent Nickel. Environmental Health Perspectives, 115(1), 93-101.
Publication 2006
Cell Lines Cells Chlorpyrifos Clone Cells Diazinon Dieldrin Equus caballus Fetal Bovine Serum Lysine Microscopy Mus Neuronal Outgrowth Parathion PC12 Cells Penicillins Pesticides Physostigmine Poly A Serum Somatostatin-Secreting Cells Streptomycin Sulfoxide, Dimethyl
3
PC12 Cell Culture and Differentiation
Cited 25 times
As described previously (Crumpton et al. 2000 (link); Qiao et al. 2003 (link); Song et al. 1998 ), PC12 cells (1721-CRL; American Type Culture Collection, Rockville, MD) were grown in RPMI-1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% inactivated horse serum (Sigma Chemical Co., St. Louis, MO), 5% fetal bovine serum (Sigma), and 50 μg/mL penicillin–streptomycin (Invitrogen); cells were incubated with 7.5% CO2 at 37°C, and the medium was changed every 2 days. Because of the clonal instability of the PC12 cell line (Fujita et al. 1989 (link)), the experiments were performed on cells that had undergone fewer than five passages, and studies were repeated several times with different batches of cells. For studies in the undifferentiated state, 3–6 × 106 cells were seeded onto 100-mm poly-l-lysine–coated plates, and 24 hr later the medium was changed to include 5 or 50 μM CPF (purity, 98%; Chem Service, West Chester, PA). CPF was dissolved in dimethyl sulfoxide (DMSO), achieving final DMSO concentrations of 0.1–1% in the culture medium, and the corresponding control samples contained equivalent DMSO concentrations. Preliminary studies were conducted to verify that these concentrations of DMSO had no effect on the measured parameters in PC12 cells (data not shown), in agreement with earlier work (Qiao et al. 2001 (link), 2003 (link); Song et al. 1998 ).
For studies during differentiation, 3.5 × 106 cells were seeded; 24 hr later the medium was changed to include 50 ng/mL NGF (Sigma), and over the ensuing week, each culture was examined under a microscope to verify the outgrowth of neurites. CPF or physostigmine (Sigma) was added either simultaneously with the addition of NGF or after a 4-day NGF pretreatment.
Jameson R.R., Seidler F.J., Qiao D, & Slotkin T.A. (2005). Chlorpyrifos Affects Phenotypic Outcomes in a Model of Mammalian Neurodevelopment: Critical Stages Targeting Differentiation in PC12 Cells. Environmental Health Perspectives, 114(5), 667-672.
Publication 2005
Cell Lines Cells Clone Cells Equus caballus Fetal Bovine Serum Lysine Microscopy Neuronal Outgrowth PC12 Cells Penicillins Physostigmine Poly A Serum Somatostatin-Secreting Cells Streptomycin Sulfoxide, Dimethyl
4
Automated Neurite Quantification with NeurphologyJ
Cited 15 times
The design aims of NeurphologyJ are as following.
1) Minimizing human intervention. It is essential to minimize the human intervention and the number of control parameters without degrading performance during batch processing. A translation of Occam's razor principle suggests that ending up with a large number of user-settable parameters is indicative of poor algorithm design [20 (link)]. An elegant image enhancement method is proposed to facilitate the determination of threshold values of segmentation.
2) Convenience of use. NeuriteTracer [14 (link)] is effective and accurate, but a pair of nuclear and neurite marker images is needed. It is more convenient if a single image of fluorescence microscopy is sufficient to measure neurite outgrowth. Only one channel per image is needed for applying NeurphologyJ.
3) Maximizing the speed. Considering the vast amount of images generated from the high-content screening, a high analyzing speed is crucial to handle such task. NeurphologyJ makes the best use of both global morphology operations of image processing and local geometric properties of lines to speed up the quantification.
4) Achieving high accuracy. There are tradeoffs between the processing speed and the accuracy. For applications in pharmacological discoveries, the ratio of neurite lengths of the treated and non-treated neurons (rather than the absolute neurite length) is the major concern. As a result, NeurphologyJ aims to achieve high coefficient correlation with manual tracing by detecting line pixels of neurites without further using linking algorithms.
5) Robustness. Image segmentation plays an important role in quantifying neuronal morphology. The techniques of local exploration and global processing are combined to deal with the staining or the illumination variation of the high-content screenings. Some settings of threshold values can be automatically derived from the histogram of enhanced neuronal images.
6) Taking advantage of the free software ImageJ. NeurphologyJ makes the best use of ImageJ commands and uses a compact set of Java modules. Being designed as a plugin of ImageJ has the benefit of easy customization for dealing with specific applications or for future expansions. Two versions of NeurphologyJ are provided, interactive and high-throughput. The interactive version is useful for optimizing the parameters for the high-throughput version.
The algorithm of NeurphologyJ consists of five parts, one image enhancement part and four morphological quantification parts. The schematic flowchart of NeurphologyJ is shown in Figure 1. The major commands used in each part and detailed descriptions are shown below.
Ho S.Y., Chao C.Y., Huang H.L., Chiu T.W., Charoenkwan P, & Hwang E. (2011). NeurphologyJ: An automatic neuronal morphology quantification method and its application in pharmacological discovery. BMC Bioinformatics, 12, 230.
Publication 2011
Homo sapiens Light Microscopy, Fluorescence Neurites Neuronal Outgrowth Neurons Screening
5
Quantifying Neuronal Axon Outgrowth Angles
Cited 7 times
For fluorescence imaging the live cortical samples were rinsed once with phosphate buffered saline (PBS) and then incubated for 30 minutes at 37°C with 50 nM Tubulin Tracker Green (Oregon Green 488 Taxol, bis-Acetate, Life Technologies, Grand Island, NY) in PBS. The samples were then rinsed twice with PBS and immersed in fresh PBS for imaging. Fluorescence images were taken using a standard Fluorescein isothiocyanate -FITC filter: excitation/emission of 495 nm/521 nm. Axon outgrowth was tracked using the NeuronJ plugin for ImageJ (http://rsbweb.nih.gov/ij). For analysis all axons were divided into segments of ∼20 µm per segment. The angle of each segment with respect to the surface direction was measured (see Fig. 1b; nanorods point in the π radians direction for all surfaces, as shown in Fig. 1a), and plotted in histograms that quantify angular axonal outgrowth for each type of surface (see below). All surfaces were imaged using an MFP3D Atomic Force Microscope (AFM), using AC mode operation and AC 160TS cantilevers (Asylum Research, Santa Barbara, CA). Surfaces were imaged both before and after neuronal culture, and no significant change in topography was observed.
Spedden E., Wiens M.R., Demirel M.C, & Staii C. (2014). Effects of Surface Asymmetry on Neuronal Growth. PLoS ONE, 9(9), e106709.
Publication 2014
Acetate Axon Cortex, Cerebral Fluorescein Fluorescein-5-isothiocyanate Fluorescence isothiocyanate Microscopy, Atomic Force Neuronal Outgrowth Neurons Oregon Green 488 carboxylic acid Phosphates Saline Solution Taxol Tubulin

Most recents protocols related to «Neuronal Outgrowth»

1
Microcontact Printing Assay for Neurite Outgrowth
Neurite outgrowth from acutely prepared dorsal root ganglia (DRG) neurons was assayed using microcontact-printed substrates. Namely, on coverslips patterned with recombinant laminin-111 (Biolamina, Stockholm, Sweden) and the NET1ΔC variants below as the test substrate. Negative silicon masters used to create stamps for microcontact printing were provided by Dr. Siegmund Schroeter (Institute of Photonic Technology, Jena, Germany) from which PDMS stamps were cast, as described103 (link). Protein printing inks were prepared containing 20 μg/ml recombinant laminin-111 or Mus musculus NET1ΔC wild type (NP_032770, aa: 24-457) or the double mutant (NP_032770, aa:1-457, R348A, R349A) in TBS containing 1 mM CaCl2. The printing ink also contained 2 µg/ml Alexa 555- conjugated goat anti-rabbit antibodies to provide a fluorescent marker for detecting printed regions. Coverslips for stamping were cleaned33 (link) and activated with oxygen plasma for 1 minute immediately prior to stamping. Stamps covered in printing ink were incubated at 37 °C for 2 hours, then rinsed with ultrapure water and dried with nitrogen. Immediately after drying, the protein was printed from the stamp onto freshly activated glass coverslips. To create a cross-patterned substrate, the test protein was always printed on the coverslip first, the coverslip was then carefully removed from the stamp, rotated through 90 degree and then the recombinant laminin-111 was printed in bands at right angles to the test protein structures, creating a grid pattern. As a positive control, recombinant laminin-111 was printed in both directions. The printed substrates were then used within 1 hour of printing.
Meier M., Gupta M., Akgül S., McDougall M., Imhof T., Nikodemus D., Reuten R., Moya-Torres A., To V., Ferens F., Heide F., Padilla-Meier G.P., Kukura P., Huang W., Gerisch B., Mörgelin M., Poole K., Antebi A., Koch M, & Stetefeld J. (2023). The dynamic nature of netrin-1 and the structural basis for glycosaminoglycan fragment-induced filament formation. Nature Communications, 14, 1226.
Publication 2023
Anti-Antibodies CD3EAP protein, human Ganglia, Spinal Goat Laminin Mice, House Neuronal Outgrowth Neurons Nitrogen Oxygen Plasma Proteins Rabbits Silicon
2
Cell Culture Maintenance and Differentiation
C2bbe1 cells (CRL-2102) were maintained in DMEM with 110 mg/L Sodium Pyruvate (Thermo 11995065) supplemented with 10% FBS (Atlas Biologicals F-0500-D), penicillin-streptomycin (Thermo 15140122) and 10 μg/mL human transferrin (Sigma-Aldrich T8158). HCA-7 cells were maintained in a 1:1 mix of DMEM and F12 (Thermo 11320033) supplemented with 10% FBS (Atlas Biologicals F-0500-D) and penicillin-streptomycin (Thermo 15140122). MDCK cells were maintained in DMEM (Thermo 11965092) supplemented with 10% Equafetal (Atlas Biologicals EF-0500-A) and penicillin-streptomycin (Thermo 15140122).
C2bbe1 LoxP cells were maintained in 20 μg/mL blasticidin (A.G. Scientific B-1247-SOL) while HCA-7 and MDCK lox cells were maintained in 10 μg/mL blasticidin. Following cre-mediated recombination with a puromycin resistance plasmid, all reporter-expressing lines were maintained in 5 μg/mL puromycin (Cayman Chemical Company 13884).
C2bbe1, HCA-7 and MDCK cells were allowed to differentiate for 7 days after plating at 100% confluency in the presence of 2 μg/mL doxycycline (Fisher Scientific AAJ6042203) when inducing transgene and/or reporter transcript expression. Differentiation can occur in dishes, plates or on 0.4 μm transwell inserts (Corning 353090). Media was changed every 3–4 days.
CAD cells were maintained in a 1:1 mix of DMEM and F12 (Thermo 11320033) supplemented with 10% Equafetal (Atlas Biologicals EF-0500-A) and penicillin-streptomycin (Thermo 15140122). CAD lox cells were received as a gift from Eugene Makeyev (Khandelia et al., 2011 (link)) and maintained in 5 μg/mL blasticidin (A.G. Scientific B-1247-SOL).
CAD cells were grown in full growth media and induced with 2 μg/mL doxycycline for 48 hr when inducing expression of transgenic constructs. Following doxycycline induction, media was replaced with serum-free media for 48 hr which induces neurite outgrowth. Doxycycline induction was continued through the differentiation period when inducing construct expression. Reporter-expressing CAD lines were maintained in 5 μg/mL puromycin (Cayman Chemical Company 13884).
All cell lines were tested for mycoplasma and found to be negative.
Goering R., Arora A., Pockalny M.C, & Taliaferro J.M. (2023). RNA localization mechanisms transcend cell morphology. eLife, 12, e80040.
Publication 2023
Animals, Transgenic Biological Factors Caimans Cell Lines Cells Culture Media Culture Media, Serum-Free Doxycycline Homo sapiens Hyperostosis, Diffuse Idiopathic Skeletal Madin Darby Canine Kidney Cells Mycoplasma Neuronal Outgrowth Penicillins Plasmids Puromycin Pyruvate Recombination, Genetic Sodium Streptomycin Transferrin Transgenes
3
Larp Gene Knockdown in CAD Cells
Mouse-specific siRNAs were ordered from IDT for each Larp target. siRNAs were transfected into cells with Lipofectamine RNAiMAX (Thermo Fisher, 13778100). Briefly, 0.5 μL of 10 μM siRNA was combined with 50 μL Optimem media and 1.5 μL RNAiMAX reagent for roughly 1.0×105 cells in one well of a 24-well plate. CAD cells were transfected in the presence of doxycycline to induce expression of reporter constructs. Media was replaced 24 hr after transfection with full growth media supplemented with 2 μg/mL doxycycline. 24 hr following media change, cells were plated on PDL-coated coverslips (if imaging) and after attaching, media was changed again for serum-free media with 2 μg/mL doxycycline to induce neurite outgrowth for 48 hr. Negative control siRNAs from IDT were used to control for effects due to transfection. For all proteins tested, knockdown of expression was confirmed by qPCR normalized to HPRT. For Larp1, immunoblots confirming protein knockdown were performed with mouse monoclonal Larp1 antibody (Santa Cruz sc-515873).
Goering R., Arora A., Pockalny M.C, & Taliaferro J.M. (2023). RNA localization mechanisms transcend cell morphology. eLife, 12, e80040.
Publication 2023
Cells Culture Media Culture Media, Serum-Free Doxycycline Immunoblotting Lipofectamine Monoclonal Antibodies Mus Neuronal Outgrowth Proteins RNA, Small Interfering Transfection
4
Quantifying Neurite Outgrowth via Sholl Analysis
For the analysis of neurite outgrowth, developing neurons at 9 DIV were replated into sterile cloning cylinders (O.D. 8 × H 8 mm2; Merck, Cat. No. CLS31668), located in the middle of Matrigel-coated glass coverslips (22 mm diameter, Cat. No. 631.0159, WDR), into six-well plates. Neurons were seeded at a density of 2500 cells/cylinder and were kept in culture accordingly to the differentiation protocol. The day after seeding, the cylinder was gently removed, allowing the radial extension of neurites by developing neurons. At 20 DIV, neurons were fixed with 3.7% paraformaldehyde in PBS for 10 min, permeabilized with ice cold 0.1% Triton X-100 in PBS and incubated in blocking buffer (5% Normal Donkey Serum; Abcam, Cat. N. ab7475) for 30 min at room temperature. Nuclear bodies and neurites were stained overnight at +4 °C with an anti-tubulin III antibody (1:1000; Abcam, Cat. No. ab18207), followed by incubation with Donkey Anti-Rabbit IgG Alexa Fluor 488 (1:1000, Abcam, Cat. No. ab150073). Images were acquired using ZEISS Axio Zoom.V16 Microscope and processed using Zeiss ZEN 2.6 Blue Microscopy Software.
To quantify axonal growth, we adapted the Sholl method of concentric rings to our cultures. Prior to processing, images were modified with ImageJ v1.5.3 (National Institutes of Health, Bethesda, MD) to eliminate fluorescence background and artefacts. For application of the Sholl analysis, we employed the ShollAnalysis ImageJ plug-in, which counts the number of intersections of neurites as a function of distance from the cell soma or explant. Immunofluorescent images were imported into ImageJ and converted in greyscale, the size scale was set accordingly, and the brightness/contrast threshold was selected manually to remove the thinner and shorter neurites emerging from the clusters. Sholl analysis was performed by selecting the centre of the neuronal body cluster as the centre of outgrowth (start radius = 0 inch) and using a step size of 0.5 inch and an end radius of 8 inch. These parameters were chosen to divide the image into concentric annuli at a radial distance of 1000 μm from each other. Intersections were counted from the border of the neuronal body cluster (set as 0 μm) outwards. Results were inputted into GraphPad Prism 9.2.0 software and statistical analysis was performed between CT and FRDA groups at each individual distance from the body cluster (two-way ANOVA with Bonferroni’s test for multiple comparisons; *P < 0.05, **P < 0.01, ***P < 0.001).
Dionisi C., Chazalon M., Rai M., Keime C., Imbault V., Communi D., Puccio H., Schiffmann S.N, & Pandolfo M. (2023). Proprioceptors-enriched neuronal cultures from induced pluripotent stem cells from Friedreich ataxia patients show altered transcriptomic and proteomic profiles, abnormal neurite extension, and impaired electrophysiological properties. Brain Communications, 5(1), fcad007.
Publication 2023
alexa fluor 488 anti-IgG Antibodies, Anti-Idiotypic Axon Buffers Cell Body Cold Temperature Equus asinus Fluorescence Fluorescent Antibody Technique Friedreich Ataxia Human Body Intersectional Framework matrigel Microscopy Neurites neuro-oncological ventral antigen 2, human Neuronal Outgrowth Neurons Nuclear Bodies paraform prisma Rabbits Radius Serum Sterility, Reproductive Triton X-100 Tubulin
5
Modulation of Peripheral Nerve Regeneration and NMJ Formation by IL-10
We assessed axon extension after acute injury and the formation of neuromuscular junctions (NMJ) at 8 weeks in Thy1-GFP rats. Rats were anesthetized with 5% isoflurane, and maintained under anesthesia with 1–2% isoflurane. Analgesia was provided by Meloxicam (2 mg/kg) pre-operatively and 24 h post-operatively. The sciatic nerve was exposed and a 10-mm silicone conduit placed between the transected nerve ends and sutured with single 10/0 sutures to create an 8 mm defect. The conduits were either filled with 0.7% agarose hydrogel, or three different concentrations of rat-specific IL10 (1 µg/mL, 3 µg/mL, 7 µg/mL, recombinant rat IL10, Peprotech, part #400-19) with 0.7% agarose as a carrier. The muscle and skin were re-apposed using 6-0 Ethilon sutures. Body temperature was maintained by an electrical heating pad placed below rat within the surgical field; body temperature and heating pad temperature were monitored to maintain physiologic body temperature. All surgeons were blinded to treatment groups during surgical procedures and tissue harvests.
To determine axon extension after injury, five groups of animals (IL10 at 1, 3, or 7 µg/mL in 0.7% agarose, unloaded agarose or empty conduit, n = 6–8 animals/group, male and female) were allowed to recover for 21 days. Animals were then anesthetized and euthanized (pentobarbital (250 mg/mL, 4 mL/kg), and the regenerative nerve bridge was harvested. Bridges were placed in a well with PBS and imaged using confocal microscope (Zeiss LSM510 confocal). One spectral window for the emission profile of GFP (488 nm) and the entire extension of the regenerative bridge was imaged using tiled, Z stack images. Axonal outgrowth from the distal stump was quantified by evaluating axonal regenerative profiles extending through contiguous regions of interest (ROI) at 250-µm intervals from the distal end of the proximal stump using BitPlane (Imaris) and calculating GFP + volumes for each ROI. Differences between groups were determined by evaluating GFP + volume and the interaction between dose and contiguous ROI in a mixed-effect model, in the same group of animals.
To determine the effect of exogenous IL10 on neuromuscular junction formation, two groups of animals (n = 8/group) underwent sciatic transection and a silicone conduit repair (10 mm) filled with 3 µg/mL IL10 in 0.7% agarose or agarose only (negative control). Animals were euthanized at 8 weeks NMJ formation in the extensor digitorum longus (EDL) muscle was assessed for terminal axons (green) innervating postsynaptic acetylcholine receptors (AChRs), labeled with rhodamine-ɑ-bungarotoxin (red) conjugated to Alexa Fluor-59454 (link). Briefly, the EDL muscles were harvested bilaterally for whole-mount confocal microscopy. The muscles were dissected into four separate muscle bellies to create thinner samples. The muscles were placed on Pyrex resin-coated dishes and stained with rhodamine alpha-bungarotoxin conjugated to Alexa Fluor-594 (2.5 µg/mL; Thermo Fischer Scientific, CA) at room temperature for 30 min to label the acetylcholine receptors in the postsynaptic membrane. The muscles were rinsed with phosphate-buffered saline and mounted under a coverslip for confocal microscopy. The concentration of alpha-bungarotoxin conjugate was validated using a dose curve in untreated GFP rats. The muscles were imaged using a Zeiss LSM510 confocal microscope. Two spectral windows using the Zeiss Zen image software were used: one using the emission profile of Alexa Fluor-594 (590 nm) and the emission profile of GFP (488 nm). To evaluate the thicker muscle samples, four equidistant z-series (stack) were taken across the muscle sample to acquire images of a minimum of 100 motor end plates. The observer was blinded to the treatment group at the time of acquisition of imaging and during image analysis. The images were used to count the number of innervated and non-innervated motor end plates; innervation was determined by the extension of a GFP-expressing axon to a positively labeled motor end plate. The proportion of innervated NMJ was standardized to each contralateral control.
Golshadi M., Claffey E.F., Grenier J.K., Miller A., Willand M., Edwards M.G., Moore T.P., Sledziona M., Gordon T., Borschel G.H, & Cheetham J. (2023). Delay modulates the immune response to nerve repair. NPJ Regenerative Medicine, 8, 12.
Publication 2023
Alexa594 alpha-Bungarotoxin Amputation Stumps Anesthesia Animals Axon Bungarotoxins Cholinergic Receptors Electricity Ethilon Females Hyperostosis, Diffuse Idiopathic Skeletal IL10 protein, human Injuries Isoflurane Males Management, Pain Meloxicam Microscopy, Confocal Motor Endplate Muscle Tissue Nerve Endings Nervousness Neuromuscular Junction Neuronal Outgrowth Operative Surgical Procedures PEGDMA Hydrogel Pentobarbital Phosphates physiology Rattus norvegicus Regeneration Resins, Plant Rhodamine rhodamine-alpha-bungarotoxin Saline Solution Sciatic Nerve Sepharose Silicones Skin Surgeons Sutures Tissue, Membrane Tissue Harvesting

Top products related to «Neuronal Outgrowth»

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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L-glutamine is an amino acid that is commonly used as a dietary supplement and in cell culture media. It serves as a source of nitrogen and supports cellular growth and metabolism.

More about "Neuronal Outgrowth"

Neuronal Outgrowth, also known as Neurite Outgrowth, is a critical process in the development and function of the nervous system.
It refers to the extension of axons and dendrites, which are the projections that neurons use to form connections with other cells.
This process is essential for the formation and refinement of neural circuits, enabling the brain and nervous system to process information and coordinate various bodily functions.
Researchers studying Neuronal Outgrowth may investigate factors that promote or inhibit this growth, such as signaling molecules, extracellular matrix components, and genetic regulators.
Understanding the mechanisms underlying Neuronal Outgrowth can provide valuable insights into nervous system development, regeneration, and the potential treatment of neurological disorders.
To optimize your Neuronal Outgrowth research, PubCompare.ai is a leading AI-driven protocol comparison platform that can help you discover the best protocols and products by easily locating relevant information from literature, pre-prints, and patents.
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In your Neuronal Outgrowth research, you may utilize various cell culture components, such as Fetal Bovine Serum (FBS), Penicillin/Streptomycin, Neurite Outgrowth Staining Kit, Lipofectamine 2000, Neurobasal medium, and Poly-L-lysine.
These tools can help you analyze and visualize the growth and development of neurons in vitro.
Additionally, software like MetaMorph can be used for image analysis and quantification of Neuronal Outgrowth.
By leveraging the insights and resources available, you can optimize your Neuronal Outgrowth research and make significant advancements in understanding the mechanisms that govern this critical process in the nervous system.