dsp 4 hydrochloride  (MedChemExpress)

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: A , Representative 30µm maximum projected confocal stack images of layers 1, 2/3, and 5 of the primary somatosensory cortex in mice sacrificed 1-, 4-, 12-, and 24-weeks following treatment with DSP4 (50mg/kg) or saline. Dopamine-β-hydroxylase (DBH)-cre x mTmG mice were used to selectively label NE axons, neuronal tissue was sliced along the sagittal plane, and the native signal was amplified through processing with antibodies raised against GFP. B , IMARIS software was used to quantify the total symbol represents the total axon surface area of a single sagittal section of an individual mouse (n=5/group) and vertical bars show the standard error. * = P < 0.05; ** = P < 0.01; *** = P < 0.001.

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: Saline, Amplification, Software

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: A , Representative 30µm maximum projected confocal stack images of layers 1, 2/3, and 5 of the primary motor cortex of mice sacrificed 1-, 4-, 12-, and 24-weeks following treatment with DSP4 (50mg/kg) or saline. Dopamine-β-hydroxylase (DBH)-cre x mTmG mice were used to selectively label NE axons, neuronal tissue was sliced along the sagittal plane, and the native signal was amplified through processing with antibodies raised against GFP. B , IMARIS software was used to quantify the total axon surface area within demonstrated in A . Each plot symbol represents the total axon surface area of a single sagittal section of an individual mouse (n=5/group) and the vertical bars show the standard error. * = P < 0.05; ** = P < 0.01; *** = P < 0.001.

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: Saline, Amplification, Software

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: A , Representative 108μm thick maximum projected stack images of layer 1 of the primary somatosensory cortex show the widespread loss and slow recovery of NE density over 16 weeks in a representative DSP4-treated mouse. In a representative saline-treated mouse, the overall axon density and individual axon location and morphology was quite stable. Specific labeling of NE axons was achieved using DBHcre x Ai14 transgenic mice. The dark, sinuous patches in these images are the shadows created by large surface blood vessels. B , (top) Population time-course data measuring total axon surface area are normalized to the value measured 1 day prior to treatment. Recovery of axon density following DSP4 treatment is dominated by new axon growth. There is almost no contribution from collateral sprouting originating from survived axons. Axons that initially survived DSP4-treatment continued to survive at a rate similar to those that were saline-treated. Vertical bars show the standard error. The number of mice imaged at each timepoint, shown in the lower panel, varied due to the deterioration of the imaging window’s clarity over time and downtime due to repair of the imaging rig from week to week. This change in the number of mice assessed at each timepoint, coupled with the variation in lesion magnitude across DSP4-treated animals (SE of Survived axons = 5.0% at week 2), led to some apparent instability across time in the population measures. However, examining survived axons within individual animals shows consistent stability over 16 weeks .

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: Saline, Labeling, Transgenic Assay, Imaging

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: A , Representative 3-by-3 tiled 30µm maximum projected confocal stack images of the locus coeruleus 24-weeks following treatment with DSP4 (50mg/kg) or saline. DBHcre x Ai14 mice were used to selectively label NE neurons with tdTomato. The brain was sliced along the sagittal plane, and the tdTomato signal was amplified using cross-reactive antibodies raised against DsRed. B , IMARIS software was used to produce an exhaustive count of the Ai14-positive cell bodies within the locus coeruleus of individual animals. No significant difference in the number of cells was detected between the saline (n=8) and DSP4 (n=9) groups (p = 0.289). The standard error is represented by vertical bars.

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: Saline, Amplification, Software

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: A , Representative 108µm 3-D reconstructed images of the primary somatosensory cortex show virally induced GCaMP8s (green) and DBHcre x Ai14 (red) one day prior to and two weeks following lesion with the NE-specific neurotoxin DSP4. Viral injection 7 weeks prior to lesion induces patchy GCaMP expression which shows the tiled patterning characteristic of astrocytes. Note that the expression pattern and intensity of GCaMP was not significantly altered over the 15 day-long span encompassing the DSP4 challenge. B , 3µm-thick maximally projected z-stack images of the same GCaMP8s signal shown in A reveals the tendril-like processes characteristic of astrocytes. C , Representative single image of layer 2/3 of the primary somatosensory cortex 16 weeks following infection with AAV5-gfaABC1D-jGCaMP8s to selectively induce GCaMP8s in cortical astrocytes. Fixed brain tissue was sliced in the sagittal plane and processed with antibodies raised against the astrocyte marker S100β. As in A and B, the GCaMP8s signal demonstrates a patchy transfection pattern and tiling while S100β labels all astrocytes, highlighting particular cytoskeletal elements. White arrowheads indicate a capillary contacted by S100β-positive astrocyte endfeet.

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: Injection, Expressing, Infection, Marker, Transfection

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: A , Schematic of the functional imaging configuration. The primary somatosensory cortex of DBHcre x Ai14 mice was infected with AAV5-gfaABC1D-jGCaMP8s (designed to drive GCaMP8s expression in astrocytes) 5-7 weeks prior to recording. The awake mouse was placed on a treadmill (not depicted) and head fixed. Compressed air (30PSI) was delivered through a tube to the back of the neck to elicit a startle-response and, following a short delay, NE release. B , Representative single trial showing a Ca 2+ transient measured in a group of adjacent astrocytes evoked by a 1s long air-puff (grey bars) to the back of the mouse’s neck. Treatment with the selective α1-adrenoreceptor antagonist prazosin (7.5mg/kg) 15 minutes prior to the trial abolishes the startle-evoked astrocytic Ca 2+ transient, indicating that it is mediated by NE release. C , Representative individual (non-averaged) false-color thermal-coded false color images of GCaMP8s fluorescence before and after treatment with 7.5mg/kg Prazosin. D-F , Select response characteristics before and after treatment with the NE-specific neurotoxin DSP4 (50mg/kg) or saline. During each assessment week, animals underwent 4 individual trials with a 5-minute inter-trial interval. Individual trials were excluded from analysis using a baseline stability criterion (see methods). Each plot point represents an individual mouse and vertical bars show the standard error. D , The proportion of trials each assessment week that elicited a startle-evoked Ca 2+ response in neocortical astrocytes. E , The peak amplitude of all GCaMP8s response traces each week. F , The latency from stimulus onset to 25% of the maximum amplitude of all GCaMP8s response traces each week. * = P < 0.05; *** = P < 0.001.

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: Functional Assay, Imaging, Infection, Expressing, Fluorescence, Saline

Journal: bioRxiv

Article Title: Functional Regrowth of Norepinephrine Axons in the Adult Mouse Brain Following Injury

doi: 10.1101/2024.08.19.608684

Figure Lengend Snippet: Long-term in vivo 2-photon imaging shows the regrowth of NE axons innervating the adult mouse somatosensory cortex following DSP4 treatment. Individual mouse (shaded) and population mean +/- standard error (solid) measurements of the axon surface area are shown. The population data are the same as illustrated in and are reproduced here to allow for comparison with the individual mouse measurements. Weeks during which data was able to be collected are indicated by shaded closed circles. Collection was sometimes limited by repair of the imaging rig and lines representing individual mice sometimes terminate due to deterioration of the optical quality of the imaging window. ** = P < 0.01; *** = P < 0.001.

Article Snippet: 4-6 weeks following viral injection and cranial window installation surgery, mice received a single 50mg/kg intraperitoneal dose of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, Tocris Bioscience #2958, PubChem ID: 38533) reconstituted in sterile water or an injection of volume-matched saline as a control.

Techniques: In Vivo, Imaging, Comparison