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    Alomone Labs ngf
    Phenotypic changes in prostatic acid phosphatase (PAP)(+) dorsal root ganglion neurons in resiniferatoxin (RTX)-induced neuropathy. (a–e) Double immunofluorescence staining was performed with anti-activating transcription factor 3 (ATF3; a–e in red) and anti-PAP (a–e in green) antisera in the vehicle (a), RTX (b), 4-methylcatechol (4MC) (c), <t>NGF</t> (RTX + <t>2.5S</t> NGF; d), and (e) abNGF (4MC + anti-NGF antisera) groups. (f–h) The graphs quantify the density changes in (f) PAP(+) and (g) ATF3(+) neurons and (h) the ratio changes in ATF3(+)/PAP(+) neurons according to Panels (a) to (e). (i) The mechanical thresholds were assessed using the up-and-down method with von Frey monofilaments, as described in the Materials and Methods. The graph indicates the changes in the mechanical thresholds in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF (dotted bar) groups. (j, k) The graphs indicate that the mechanical thresholds correlated with PAP expression, that is, a linear correlation with (j) PAP densities and (k) an inverse correlation with the ratio of ATF3(+)/PAP(+) neurons. (l, m) The graphs show the diameter histogram of ATF3(+)/PAP(+) neurons in the RTX (l) and 4MC (M) groups. The diameter histogram shows no difference between the RTX and 4MC groups, but higher numbers of ATF3(+)/PAP(+) neurons in the RTX group. Bar, 50 µm. * p
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    Images

    1) Product Images from "NGF-trkA signaling modulates the analgesic effects of prostatic acid phosphatase in resiniferatoxin-induced neuropathy"

    Article Title: NGF-trkA signaling modulates the analgesic effects of prostatic acid phosphatase in resiniferatoxin-induced neuropathy

    Journal: Molecular Pain

    doi: 10.1177/1744806916656846

    Phenotypic changes in prostatic acid phosphatase (PAP)(+) dorsal root ganglion neurons in resiniferatoxin (RTX)-induced neuropathy. (a–e) Double immunofluorescence staining was performed with anti-activating transcription factor 3 (ATF3; a–e in red) and anti-PAP (a–e in green) antisera in the vehicle (a), RTX (b), 4-methylcatechol (4MC) (c), NGF (RTX + 2.5S NGF; d), and (e) abNGF (4MC + anti-NGF antisera) groups. (f–h) The graphs quantify the density changes in (f) PAP(+) and (g) ATF3(+) neurons and (h) the ratio changes in ATF3(+)/PAP(+) neurons according to Panels (a) to (e). (i) The mechanical thresholds were assessed using the up-and-down method with von Frey monofilaments, as described in the Materials and Methods. The graph indicates the changes in the mechanical thresholds in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF (dotted bar) groups. (j, k) The graphs indicate that the mechanical thresholds correlated with PAP expression, that is, a linear correlation with (j) PAP densities and (k) an inverse correlation with the ratio of ATF3(+)/PAP(+) neurons. (l, m) The graphs show the diameter histogram of ATF3(+)/PAP(+) neurons in the RTX (l) and 4MC (M) groups. The diameter histogram shows no difference between the RTX and 4MC groups, but higher numbers of ATF3(+)/PAP(+) neurons in the RTX group. Bar, 50 µm. * p
    Figure Legend Snippet: Phenotypic changes in prostatic acid phosphatase (PAP)(+) dorsal root ganglion neurons in resiniferatoxin (RTX)-induced neuropathy. (a–e) Double immunofluorescence staining was performed with anti-activating transcription factor 3 (ATF3; a–e in red) and anti-PAP (a–e in green) antisera in the vehicle (a), RTX (b), 4-methylcatechol (4MC) (c), NGF (RTX + 2.5S NGF; d), and (e) abNGF (4MC + anti-NGF antisera) groups. (f–h) The graphs quantify the density changes in (f) PAP(+) and (g) ATF3(+) neurons and (h) the ratio changes in ATF3(+)/PAP(+) neurons according to Panels (a) to (e). (i) The mechanical thresholds were assessed using the up-and-down method with von Frey monofilaments, as described in the Materials and Methods. The graph indicates the changes in the mechanical thresholds in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF (dotted bar) groups. (j, k) The graphs indicate that the mechanical thresholds correlated with PAP expression, that is, a linear correlation with (j) PAP densities and (k) an inverse correlation with the ratio of ATF3(+)/PAP(+) neurons. (l, m) The graphs show the diameter histogram of ATF3(+)/PAP(+) neurons in the RTX (l) and 4MC (M) groups. The diameter histogram shows no difference between the RTX and 4MC groups, but higher numbers of ATF3(+)/PAP(+) neurons in the RTX group. Bar, 50 µm. * p

    Techniques Used: Double Immunofluorescence Staining, Expressing

    Colocalization of prostatic acid phosphatase (PAP) and high-affinity nerve growth factor (trkA) receptor after resiniferatoxin (RTX)-induced neuropathy. (a–j) Double immunofluorescence staining of dorsal root ganglia sections was performed with anti-PAP (a–e) and trkA (f-j) in the vehicle (a, f), RTX (b, g), 4-methylcatechol (4MC; c, h), (d, i) NGF (RTX + 2.5S NGF), and (e, j) abNGF (4MC + anti-NGF antisera) groups. (k, l) The graphs show the changes in (k) trkA density and (l) the colocalized ratios of PAP(+)/trkA(+) neurons in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF groups (dotted bar) according to Panels (a) to (j). (m) The graph shows that the mechanical thresholds correlate linearly with the ratios of PAP(+)/trkA(+) neurons. Bar, 50 µm. * p
    Figure Legend Snippet: Colocalization of prostatic acid phosphatase (PAP) and high-affinity nerve growth factor (trkA) receptor after resiniferatoxin (RTX)-induced neuropathy. (a–j) Double immunofluorescence staining of dorsal root ganglia sections was performed with anti-PAP (a–e) and trkA (f-j) in the vehicle (a, f), RTX (b, g), 4-methylcatechol (4MC; c, h), (d, i) NGF (RTX + 2.5S NGF), and (e, j) abNGF (4MC + anti-NGF antisera) groups. (k, l) The graphs show the changes in (k) trkA density and (l) the colocalized ratios of PAP(+)/trkA(+) neurons in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF groups (dotted bar) according to Panels (a) to (j). (m) The graph shows that the mechanical thresholds correlate linearly with the ratios of PAP(+)/trkA(+) neurons. Bar, 50 µm. * p

    Techniques Used: Double Immunofluorescence Staining

    2) Product Images from "Ca2+ Signalling Induced by NGF Identifies a Subset of Capsaicin-Excitable Neurons Displaying Enhanced Chemo-Nociception in Dorsal Root Ganglion Explants from Adult pirt-GCaMP3 Mouse"

    Article Title: Ca2+ Signalling Induced by NGF Identifies a Subset of Capsaicin-Excitable Neurons Displaying Enhanced Chemo-Nociception in Dorsal Root Ganglion Explants from Adult pirt-GCaMP3 Mouse

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms22052589

    NGF attenuates CAPS tachyphylaxis. Response ratios (peak value obtained for second CAPS stimulation after [A]/peak value for first stimulation before [B] exposure to 100 ng/mL NGF) for the ( A ) number of responders or ( B – D ) Max. signal intensity evoked during the second exposure to 0.3 (blue) or 1 μM CAPS (red) divided by the values elicited the first time when the DRG were stimulated with the same concentration of agonist. In ( B – D ), the calculations were restricted to cells that conformed to the following criteria: ( B ) neurons that produced above threshold signals to 0.3 or 1 μM CAPS the first time each concentration was applied; note that both above and below threshold values elicited by the second stimulation with either agonist concentration were included in these analyses, ( C ) neurons excited above threshold both times the DRG were exposed to 0.3 or 1 μM CAPS, ( D ) all DRGNs excited at least one time by either the first or second stimulation with 0.3 or 1 μM CAPS; although the threshold criterion was used to identify the neurons to analyse, comparisons before and after NGF included values both above and below threshold. Paired two-tailed Student’s t-test were performed to compare the mean values of [A] and [B]; * p
    Figure Legend Snippet: NGF attenuates CAPS tachyphylaxis. Response ratios (peak value obtained for second CAPS stimulation after [A]/peak value for first stimulation before [B] exposure to 100 ng/mL NGF) for the ( A ) number of responders or ( B – D ) Max. signal intensity evoked during the second exposure to 0.3 (blue) or 1 μM CAPS (red) divided by the values elicited the first time when the DRG were stimulated with the same concentration of agonist. In ( B – D ), the calculations were restricted to cells that conformed to the following criteria: ( B ) neurons that produced above threshold signals to 0.3 or 1 μM CAPS the first time each concentration was applied; note that both above and below threshold values elicited by the second stimulation with either agonist concentration were included in these analyses, ( C ) neurons excited above threshold both times the DRG were exposed to 0.3 or 1 μM CAPS, ( D ) all DRGNs excited at least one time by either the first or second stimulation with 0.3 or 1 μM CAPS; although the threshold criterion was used to identify the neurons to analyse, comparisons before and after NGF included values both above and below threshold. Paired two-tailed Student’s t-test were performed to compare the mean values of [A] and [B]; * p

    Techniques Used: Concentration Assay, Produced, Two Tailed Test

    Brief exposure to NGF results in sensitisation to CAPS. Analysis tools were applied to signals evoked by 0.3 (blue dots) and 1 μM CAPS (red dots) before [B] and after [A] 5 min exposure to 100 ng/mL NGF (blue trace in Figure 1 A) to measure ( A ) Max. intensity, ( B ) signal duration and ( C ) lag time. Asterisks represent significance between the measurements before and after NGF treatment, determined by Student’s t-test for unpaired samples, unequal variance; * p
    Figure Legend Snippet: Brief exposure to NGF results in sensitisation to CAPS. Analysis tools were applied to signals evoked by 0.3 (blue dots) and 1 μM CAPS (red dots) before [B] and after [A] 5 min exposure to 100 ng/mL NGF (blue trace in Figure 1 A) to measure ( A ) Max. intensity, ( B ) signal duration and ( C ) lag time. Asterisks represent significance between the measurements before and after NGF treatment, determined by Student’s t-test for unpaired samples, unequal variance; * p

    Techniques Used:

    NGF evokes signals if applied before CAPS and excites a new cohort when re-applied afterwards; CAPS-induced signal strength correlates with responsiveness to NGF. ( A ) Summed increase in fluorescence from DRG treated, in the following order, with 100 ng/mL NGF (20 min followed by 40 min washout, [N1]); 1 μM CAPS (5 min, 15 min washout, [C1]); NGF for a second time (20 min, 20 min washout, [N2]); 1 μM CAPS again (5 min, 15 min washout, [C2]) and, finally, 10 μM CAPS (5 min, 35 min washout, [C3]). ( B ) Shows the relative increase in the number of active cells (number activated by 10 μM CAPS [C3]/number excited by 1 μM [C1]) calculated for all CAPS-excitable cells (Total) or only the subsets that were NGF-excitable and -refractory cells. ( C – E ) Neurons that responded during the first ([C1]; blue dots) or second exposure ([C2]; green dots) to 1 μM CAPS and those excited by 10 μM CAPS ([C3]; red dots) were sub-categorised according to whether they were excited both times (2, abscissa) on exposure to NGF (i.e., during [N1] and [N2]), only once (1, abscissa) by either application (i.e., [N1] or [N2]) or remained inactive both times (0, abscissa). Each sub-category was analysed for ( C ) lag time, ( D ) signal duration and ( E ) Max. intensity. Plotted data represents mean ± s.e.m. Asterisks indicate p values for Student’s t -tests compared between group 1 and the requisite data set in group 0, or between groups 1 and 2. ** p
    Figure Legend Snippet: NGF evokes signals if applied before CAPS and excites a new cohort when re-applied afterwards; CAPS-induced signal strength correlates with responsiveness to NGF. ( A ) Summed increase in fluorescence from DRG treated, in the following order, with 100 ng/mL NGF (20 min followed by 40 min washout, [N1]); 1 μM CAPS (5 min, 15 min washout, [C1]); NGF for a second time (20 min, 20 min washout, [N2]); 1 μM CAPS again (5 min, 15 min washout, [C2]) and, finally, 10 μM CAPS (5 min, 35 min washout, [C3]). ( B ) Shows the relative increase in the number of active cells (number activated by 10 μM CAPS [C3]/number excited by 1 μM [C1]) calculated for all CAPS-excitable cells (Total) or only the subsets that were NGF-excitable and -refractory cells. ( C – E ) Neurons that responded during the first ([C1]; blue dots) or second exposure ([C2]; green dots) to 1 μM CAPS and those excited by 10 μM CAPS ([C3]; red dots) were sub-categorised according to whether they were excited both times (2, abscissa) on exposure to NGF (i.e., during [N1] and [N2]), only once (1, abscissa) by either application (i.e., [N1] or [N2]) or remained inactive both times (0, abscissa). Each sub-category was analysed for ( C ) lag time, ( D ) signal duration and ( E ) Max. intensity. Plotted data represents mean ± s.e.m. Asterisks indicate p values for Student’s t -tests compared between group 1 and the requisite data set in group 0, or between groups 1 and 2. ** p

    Techniques Used: Fluorescence

    NGF evokes Ca2+ signals in a subset of CAPS-excitable cells. ( A ) Traces show increases in fluorescence intensity (summed from 3 experiments) in DRGNs exposed sequentially to CAPS for 5 min. (black bars, at concentrations indicated) and 100 ng/mL NGF for 5 (blue) or 20 min. (red). In the latter case, a 5 min. washout after the 20 min. exposure to NGF has been omitted from the figure to keep the subsequent responses to CAPS in register with those in the blue trace. Note that F 0 was re-zeroed (see Section 4 ) before each addition of CAPS or NGF. Examples of fluorescence traces from individual DRGNs are presented in Supplementary Figure S1 . ( B ) Shows the mean ± s.e.m. Max. fluorescence intensity increases in neurons that responded above threshold upon exposure to each stimulus, as indicated on abscissa, in DRG exposed to NGF for 5 (blue bars) or 20 min. (red bars). Significant differences were observed for 0.3 μM CAPS before NGF ( p = 0.03), reflecting minor variation between experimental groups, and for 1 μM CAPS after NGF ( p = 0.02) but this latter difference might be due to the high background fluorescence after 20 min. with NGF, which may interfere with the quantification of subsequent responses to CAPS. ( C ) Number of active cells counted in 1 min. intervals after the addition of NGF, plotted against time; data summed from 3 independent recordings for each NGF treatment. ( D ) Mean number of cells activated (± s.e.m., N = 3) over 20 min. after the addition of CAPS (at concentrations indicated), or NGF, in DRG exposed to the latter for 5 (blue) or 20 min. (red). Results obtained with either CAPS concentration are shown for a two-tailed Student’s t -test between groups exposed to NGF for 5 min. and those treated for 20 min; * p
    Figure Legend Snippet: NGF evokes Ca2+ signals in a subset of CAPS-excitable cells. ( A ) Traces show increases in fluorescence intensity (summed from 3 experiments) in DRGNs exposed sequentially to CAPS for 5 min. (black bars, at concentrations indicated) and 100 ng/mL NGF for 5 (blue) or 20 min. (red). In the latter case, a 5 min. washout after the 20 min. exposure to NGF has been omitted from the figure to keep the subsequent responses to CAPS in register with those in the blue trace. Note that F 0 was re-zeroed (see Section 4 ) before each addition of CAPS or NGF. Examples of fluorescence traces from individual DRGNs are presented in Supplementary Figure S1 . ( B ) Shows the mean ± s.e.m. Max. fluorescence intensity increases in neurons that responded above threshold upon exposure to each stimulus, as indicated on abscissa, in DRG exposed to NGF for 5 (blue bars) or 20 min. (red bars). Significant differences were observed for 0.3 μM CAPS before NGF ( p = 0.03), reflecting minor variation between experimental groups, and for 1 μM CAPS after NGF ( p = 0.02) but this latter difference might be due to the high background fluorescence after 20 min. with NGF, which may interfere with the quantification of subsequent responses to CAPS. ( C ) Number of active cells counted in 1 min. intervals after the addition of NGF, plotted against time; data summed from 3 independent recordings for each NGF treatment. ( D ) Mean number of cells activated (± s.e.m., N = 3) over 20 min. after the addition of CAPS (at concentrations indicated), or NGF, in DRG exposed to the latter for 5 (blue) or 20 min. (red). Results obtained with either CAPS concentration are shown for a two-tailed Student’s t -test between groups exposed to NGF for 5 min. and those treated for 20 min; * p

    Techniques Used: Fluorescence, Concentration Assay, Two Tailed Test

    Acute sensitisation by NGF of responses to CAPS is most prevalent in cells that exhibit [Ca2+]i signals when the neurotrophin is applied. Neurons that responded to 1 μM CAPS both times when it was applied before and after 5 min exposure to NGF (blue trace in Figure 1 A) were split into two groups depending on whether they additionally exhibited Ca 2+ signals in response to NGF alone (red dots, n = 13 neurons from 3 DRG recordings) or did not (blue dots, n = 170). The analysis tools were applied to measure ( A ) Max. intensity, ( B ) signal duration and ( C ) lag time for the CAPS-evoked signals before [B] (on abscissa) and after [A] the treatment with NGF. Asterisks show significant differences within groups between responses before and after NGF treatment (Student’s t -test, paired samples; * p
    Figure Legend Snippet: Acute sensitisation by NGF of responses to CAPS is most prevalent in cells that exhibit [Ca2+]i signals when the neurotrophin is applied. Neurons that responded to 1 μM CAPS both times when it was applied before and after 5 min exposure to NGF (blue trace in Figure 1 A) were split into two groups depending on whether they additionally exhibited Ca 2+ signals in response to NGF alone (red dots, n = 13 neurons from 3 DRG recordings) or did not (blue dots, n = 170). The analysis tools were applied to measure ( A ) Max. intensity, ( B ) signal duration and ( C ) lag time for the CAPS-evoked signals before [B] (on abscissa) and after [A] the treatment with NGF. Asterisks show significant differences within groups between responses before and after NGF treatment (Student’s t -test, paired samples; * p

    Techniques Used:

    NGF-excitable cells exhibit more robust responses to CAPS. Further analysis was performed on DRG subjected to the protocol described for the red trace in Figure 1 A. Cells were categorised according to whether or not they responded to 20 min. with NGF. ( A ) The number of NGF-excitable (red bars) and -refractory (blue bars) cells that responded to each treatment with the indicated CAPS concentrations applied before [B] or after [A] NGF. Analysis tools were applied to each sub-category to measure ( B ) Lag, ( C ) Duration and ( D ) Max. intensity, in all cases plotted as means ± s.e.m., with blue and red dots representing, respectively, values from individual cells assigned to the NGF-refractory and -excitable categories. Black asterisks represent p values derived by unpaired Student’s t-test between the NGF-excitable and –refractory cells within each CAPS stimulation group. Coloured asterisks show results from paired Student’s t-test for measurements of signals evoked by 1 μM CAPS before and after NGF, respectively, for each cell category; *** p
    Figure Legend Snippet: NGF-excitable cells exhibit more robust responses to CAPS. Further analysis was performed on DRG subjected to the protocol described for the red trace in Figure 1 A. Cells were categorised according to whether or not they responded to 20 min. with NGF. ( A ) The number of NGF-excitable (red bars) and -refractory (blue bars) cells that responded to each treatment with the indicated CAPS concentrations applied before [B] or after [A] NGF. Analysis tools were applied to each sub-category to measure ( B ) Lag, ( C ) Duration and ( D ) Max. intensity, in all cases plotted as means ± s.e.m., with blue and red dots representing, respectively, values from individual cells assigned to the NGF-refractory and -excitable categories. Black asterisks represent p values derived by unpaired Student’s t-test between the NGF-excitable and –refractory cells within each CAPS stimulation group. Coloured asterisks show results from paired Student’s t-test for measurements of signals evoked by 1 μM CAPS before and after NGF, respectively, for each cell category; *** p

    Techniques Used: Derivative Assay

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    Alomone Labs mouse ngf
    Effect of kCer molecular species on neurite outgrowth, neurite length distribution, and cell morphological changes in <t>NGF-primed</t> PC12 cells. ( A ) Representative images of NGF (100 ng/mL, 2.8 nM in medium)-primed PC12 cells (7.5 × 10 4 per mL) incubated in the absence or presence of NGF and 50 µM kCer or kCer molecular species (d18:2 4t,8t -C16h:0, d18:2 4t,8c -C16h:0, d18:2 4t,8t -C18h:0, d18:2 4t,8c -C18h:0, d18:2 4t,8t -C20h:0, d18:2 4t,8c -C20h:0, <t>t18:1</t> 8c -C22h:0, t18:1 8c -C24h:0). Cells were stained by 1% CBB and photographed (20× magnification). Scale bar: 100 μm. ( B ) Neurite length distribution changes of PC12 cells incubated with 50 µM kCer or kCer molecular species (t18:1 8c -C22h:0, t18:1 8c -C24h:0, d18:2 4t,8t -C16h:0, d18:2 4t,8c -C16h:0, d18:2 4t,8t -C18h:0, d18:2 4t,8c -C18h:0, d18:2 4t,8t -C20h:0, d18:2 4t,8c -C20h:0). All counting of the number of neurites with lengths of 10 to 150 μm is shown as 100% and calculated for each of x-axis valued neurite length.
    Mouse Ngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    Effect of kCer molecular species on neurite outgrowth, neurite length distribution, and cell morphological changes in NGF-primed PC12 cells. ( A ) Representative images of NGF (100 ng/mL, 2.8 nM in medium)-primed PC12 cells (7.5 × 10 4 per mL) incubated in the absence or presence of NGF and 50 µM kCer or kCer molecular species (d18:2 4t,8t -C16h:0, d18:2 4t,8c -C16h:0, d18:2 4t,8t -C18h:0, d18:2 4t,8c -C18h:0, d18:2 4t,8t -C20h:0, d18:2 4t,8c -C20h:0, t18:1 8c -C22h:0, t18:1 8c -C24h:0). Cells were stained by 1% CBB and photographed (20× magnification). Scale bar: 100 μm. ( B ) Neurite length distribution changes of PC12 cells incubated with 50 µM kCer or kCer molecular species (t18:1 8c -C22h:0, t18:1 8c -C24h:0, d18:2 4t,8t -C16h:0, d18:2 4t,8c -C16h:0, d18:2 4t,8t -C18h:0, d18:2 4t,8c -C18h:0, d18:2 4t,8t -C20h:0, d18:2 4t,8c -C20h:0). All counting of the number of neurites with lengths of 10 to 150 μm is shown as 100% and calculated for each of x-axis valued neurite length.

    Journal: International Journal of Molecular Sciences

    Article Title: Neurite Outgrowth and Morphological Changes Induced by 8-trans Unsaturation of Sphingadienine in kCer Molecular Species

    doi: 10.3390/ijms20092116

    Figure Lengend Snippet: Effect of kCer molecular species on neurite outgrowth, neurite length distribution, and cell morphological changes in NGF-primed PC12 cells. ( A ) Representative images of NGF (100 ng/mL, 2.8 nM in medium)-primed PC12 cells (7.5 × 10 4 per mL) incubated in the absence or presence of NGF and 50 µM kCer or kCer molecular species (d18:2 4t,8t -C16h:0, d18:2 4t,8c -C16h:0, d18:2 4t,8t -C18h:0, d18:2 4t,8c -C18h:0, d18:2 4t,8t -C20h:0, d18:2 4t,8c -C20h:0, t18:1 8c -C22h:0, t18:1 8c -C24h:0). Cells were stained by 1% CBB and photographed (20× magnification). Scale bar: 100 μm. ( B ) Neurite length distribution changes of PC12 cells incubated with 50 µM kCer or kCer molecular species (t18:1 8c -C22h:0, t18:1 8c -C24h:0, d18:2 4t,8t -C16h:0, d18:2 4t,8c -C16h:0, d18:2 4t,8t -C18h:0, d18:2 4t,8c -C18h:0, d18:2 4t,8t -C20h:0, d18:2 4t,8c -C20h:0). All counting of the number of neurites with lengths of 10 to 150 μm is shown as 100% and calculated for each of x-axis valued neurite length.

    Article Snippet: General The following materials were commercially obtained: the kGlcCer and kGlcCer molecular species d18:24t,8c -C16h:0, d18:24t,8t -C16h:0, d18:24t,8c -C18h:0, d18:24t,8t -C18h:0, d18:24t,8c -C20h:0, d18:24t,8t -C20h:0, t18:18c -C22h:0, t18:18c -C23h:0, and t18:18c -C24h:0 were purchased from Nagara Science Co., Ltd. (Gifu, Japan); 2.5S mouse NGF (N100NF4325) was from Alomone Labs, Jerusalem, Israel; semaphorin 3A (Sema3A, 193-17051) was from Wako Corp. (Tokyo, Japan); anti-CRMP2 pAb and anti-phospho-CRMP2 pAb (pthr509) were from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Incubation, Staining

    Phenotypic changes in prostatic acid phosphatase (PAP)(+) dorsal root ganglion neurons in resiniferatoxin (RTX)-induced neuropathy. (a–e) Double immunofluorescence staining was performed with anti-activating transcription factor 3 (ATF3; a–e in red) and anti-PAP (a–e in green) antisera in the vehicle (a), RTX (b), 4-methylcatechol (4MC) (c), NGF (RTX + 2.5S NGF; d), and (e) abNGF (4MC + anti-NGF antisera) groups. (f–h) The graphs quantify the density changes in (f) PAP(+) and (g) ATF3(+) neurons and (h) the ratio changes in ATF3(+)/PAP(+) neurons according to Panels (a) to (e). (i) The mechanical thresholds were assessed using the up-and-down method with von Frey monofilaments, as described in the Materials and Methods. The graph indicates the changes in the mechanical thresholds in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF (dotted bar) groups. (j, k) The graphs indicate that the mechanical thresholds correlated with PAP expression, that is, a linear correlation with (j) PAP densities and (k) an inverse correlation with the ratio of ATF3(+)/PAP(+) neurons. (l, m) The graphs show the diameter histogram of ATF3(+)/PAP(+) neurons in the RTX (l) and 4MC (M) groups. The diameter histogram shows no difference between the RTX and 4MC groups, but higher numbers of ATF3(+)/PAP(+) neurons in the RTX group. Bar, 50 µm. * p

    Journal: Molecular Pain

    Article Title: NGF-trkA signaling modulates the analgesic effects of prostatic acid phosphatase in resiniferatoxin-induced neuropathy

    doi: 10.1177/1744806916656846

    Figure Lengend Snippet: Phenotypic changes in prostatic acid phosphatase (PAP)(+) dorsal root ganglion neurons in resiniferatoxin (RTX)-induced neuropathy. (a–e) Double immunofluorescence staining was performed with anti-activating transcription factor 3 (ATF3; a–e in red) and anti-PAP (a–e in green) antisera in the vehicle (a), RTX (b), 4-methylcatechol (4MC) (c), NGF (RTX + 2.5S NGF; d), and (e) abNGF (4MC + anti-NGF antisera) groups. (f–h) The graphs quantify the density changes in (f) PAP(+) and (g) ATF3(+) neurons and (h) the ratio changes in ATF3(+)/PAP(+) neurons according to Panels (a) to (e). (i) The mechanical thresholds were assessed using the up-and-down method with von Frey monofilaments, as described in the Materials and Methods. The graph indicates the changes in the mechanical thresholds in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF (dotted bar) groups. (j, k) The graphs indicate that the mechanical thresholds correlated with PAP expression, that is, a linear correlation with (j) PAP densities and (k) an inverse correlation with the ratio of ATF3(+)/PAP(+) neurons. (l, m) The graphs show the diameter histogram of ATF3(+)/PAP(+) neurons in the RTX (l) and 4MC (M) groups. The diameter histogram shows no difference between the RTX and 4MC groups, but higher numbers of ATF3(+)/PAP(+) neurons in the RTX group. Bar, 50 µm. * p

    Article Snippet: Briefly, 2.5S NGF (Alomone Labs, Jerusalem, Israel) was dissolved in Dulbecco’s Modified Eagle Medium, and mice received NGF (1 µg/10 g) immediately after RTX treatment (the NGF group).

    Techniques: Double Immunofluorescence Staining, Expressing

    Colocalization of prostatic acid phosphatase (PAP) and high-affinity nerve growth factor (trkA) receptor after resiniferatoxin (RTX)-induced neuropathy. (a–j) Double immunofluorescence staining of dorsal root ganglia sections was performed with anti-PAP (a–e) and trkA (f-j) in the vehicle (a, f), RTX (b, g), 4-methylcatechol (4MC; c, h), (d, i) NGF (RTX + 2.5S NGF), and (e, j) abNGF (4MC + anti-NGF antisera) groups. (k, l) The graphs show the changes in (k) trkA density and (l) the colocalized ratios of PAP(+)/trkA(+) neurons in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF groups (dotted bar) according to Panels (a) to (j). (m) The graph shows that the mechanical thresholds correlate linearly with the ratios of PAP(+)/trkA(+) neurons. Bar, 50 µm. * p

    Journal: Molecular Pain

    Article Title: NGF-trkA signaling modulates the analgesic effects of prostatic acid phosphatase in resiniferatoxin-induced neuropathy

    doi: 10.1177/1744806916656846

    Figure Lengend Snippet: Colocalization of prostatic acid phosphatase (PAP) and high-affinity nerve growth factor (trkA) receptor after resiniferatoxin (RTX)-induced neuropathy. (a–j) Double immunofluorescence staining of dorsal root ganglia sections was performed with anti-PAP (a–e) and trkA (f-j) in the vehicle (a, f), RTX (b, g), 4-methylcatechol (4MC; c, h), (d, i) NGF (RTX + 2.5S NGF), and (e, j) abNGF (4MC + anti-NGF antisera) groups. (k, l) The graphs show the changes in (k) trkA density and (l) the colocalized ratios of PAP(+)/trkA(+) neurons in the vehicle (open bar), RTX (filled bar), 4MC (grey bar), NGF (slashed bar), and abNGF groups (dotted bar) according to Panels (a) to (j). (m) The graph shows that the mechanical thresholds correlate linearly with the ratios of PAP(+)/trkA(+) neurons. Bar, 50 µm. * p

    Article Snippet: Briefly, 2.5S NGF (Alomone Labs, Jerusalem, Israel) was dissolved in Dulbecco’s Modified Eagle Medium, and mice received NGF (1 µg/10 g) immediately after RTX treatment (the NGF group).

    Techniques: Double Immunofluorescence Staining

    EGF and FGF stimulation of neuroblastoma cells causes a decrease in the amount of LIFR NBFL cells were treated with 100 ng/mL of CNTF (C), OsM (O), LIF (L), FGF (F) or NGF (N) or 10 ng/mL EGF (E) or left untreated (-). After 5 h the cells were collected and LIFR and TfR levels measured by western blot analysis of lysates (a). Bands were quantified and LIFR measurements were normalized to the levels of TfR to calculate the percentage change in LIFR amounts following stimulation (b). Data in graph are averages ± SD of 3 separate experiments. P-values were calculated by ANOVA followed by a Dunnett post-test, comparing individual cytokine or growth factor treatments to untreated cells. *p

    Journal: Journal of neurochemistry

    Article Title: Transregulation of Leukemia Inhibitor Factor Receptor Expression and Function by Growth Factors in Neuroblastoma Cells

    doi: 10.1111/j.1471-4159.2008.05535.x

    Figure Lengend Snippet: EGF and FGF stimulation of neuroblastoma cells causes a decrease in the amount of LIFR NBFL cells were treated with 100 ng/mL of CNTF (C), OsM (O), LIF (L), FGF (F) or NGF (N) or 10 ng/mL EGF (E) or left untreated (-). After 5 h the cells were collected and LIFR and TfR levels measured by western blot analysis of lysates (a). Bands were quantified and LIFR measurements were normalized to the levels of TfR to calculate the percentage change in LIFR amounts following stimulation (b). Data in graph are averages ± SD of 3 separate experiments. P-values were calculated by ANOVA followed by a Dunnett post-test, comparing individual cytokine or growth factor treatments to untreated cells. *p

    Article Snippet: Human LIF and mouse nerve growth factor (NGF) were purchased from Alomone Labs (Jerusalem, Israel).

    Techniques: Western Blot

    Decreases in LIFR are correlated with robust and extended activation of Erk1/2 NBFL cells were stimulated with 100 ng/mL of CNTF, OsM, LIF, FGF or NGF or 10 ng/mL EGF for times indicated. Cells were collected and phospho-Erk1/2 (P-Erk1/2) and total Erk1/2 were measured by western blot analysis (a). Other cells were pretreated with 50 μM PD98059 for 30 minutes before stimulating with 10 ng/mL EGF for indicated times. Cells were collected and lysates blotted for P-Erk1/2 and total Erk1/2 (b) or LIFR and TfR (c). Blots shown are representative of 3 separate experiments (a and c) or 2 separate experiments (b). Data represented by the blot in 2c were quantified and are shown in graph (d). Graph shows mean ± SEM of LIFR levels following treatment with DMSO alone (closed bar), DMSO + 5h EGF (darkly shaded bar), PD98059 alone (open bar) or PD98059 + 5h EGF (lightly shaded bar). P-values were calculated by ANOVA followed by a Student-Newman-Keuls post-test. *p

    Journal: Journal of neurochemistry

    Article Title: Transregulation of Leukemia Inhibitor Factor Receptor Expression and Function by Growth Factors in Neuroblastoma Cells

    doi: 10.1111/j.1471-4159.2008.05535.x

    Figure Lengend Snippet: Decreases in LIFR are correlated with robust and extended activation of Erk1/2 NBFL cells were stimulated with 100 ng/mL of CNTF, OsM, LIF, FGF or NGF or 10 ng/mL EGF for times indicated. Cells were collected and phospho-Erk1/2 (P-Erk1/2) and total Erk1/2 were measured by western blot analysis (a). Other cells were pretreated with 50 μM PD98059 for 30 minutes before stimulating with 10 ng/mL EGF for indicated times. Cells were collected and lysates blotted for P-Erk1/2 and total Erk1/2 (b) or LIFR and TfR (c). Blots shown are representative of 3 separate experiments (a and c) or 2 separate experiments (b). Data represented by the blot in 2c were quantified and are shown in graph (d). Graph shows mean ± SEM of LIFR levels following treatment with DMSO alone (closed bar), DMSO + 5h EGF (darkly shaded bar), PD98059 alone (open bar) or PD98059 + 5h EGF (lightly shaded bar). P-values were calculated by ANOVA followed by a Student-Newman-Keuls post-test. *p

    Article Snippet: Human LIF and mouse nerve growth factor (NGF) were purchased from Alomone Labs (Jerusalem, Israel).

    Techniques: Activation Assay, Western Blot

    Effects of AUR on ERK1/2 activation and cAMP response element-binding protein (CREB) activation in PC12 cells. ( A , B ) Cells were treated with various concentrations (0, 3, and 30 μM) of AUR for 30 min or nerve growth factor (NGF, 50 ng/mL) for 10 min, and cell lysates were then prepared and subjected to immunoblot analysis; ( C ) Cells were left untreated or were pretreated for 30 min with 10 μM U0126, a MEK inhibitor, or with 10 μM H89, a PKA inhibitor, and then incubated for 30 min with 30 μM AUR. Thereafter, cells were subjected to immunoblot analysis.

    Journal: International Journal of Molecular Sciences

    Article Title: Neurotrophic Effect of Citrus Auraptene: Neuritogenic Activity in PC12 Cells

    doi: 10.3390/ijms13055338

    Figure Lengend Snippet: Effects of AUR on ERK1/2 activation and cAMP response element-binding protein (CREB) activation in PC12 cells. ( A , B ) Cells were treated with various concentrations (0, 3, and 30 μM) of AUR for 30 min or nerve growth factor (NGF, 50 ng/mL) for 10 min, and cell lysates were then prepared and subjected to immunoblot analysis; ( C ) Cells were left untreated or were pretreated for 30 min with 10 μM U0126, a MEK inhibitor, or with 10 μM H89, a PKA inhibitor, and then incubated for 30 min with 30 μM AUR. Thereafter, cells were subjected to immunoblot analysis.

    Article Snippet: Mouse NGF 2.5S (Grade I) was purchased from Alomone Labs Ltd. (Jerusalem, Israel).

    Techniques: Activation Assay, Binding Assay, Incubation

    Effects of AUR on neurite outgrowth from PC12 cells. Cells were treated with various concentrations (0, 10, 30, and 50 μM) of AUR or NGF (50 ng/mL) for 24 h, after which phase-contrast photomicrographs of the cells were taken ( A ): a , control; b , treatment with NGF; c , treatment with 30 μM AUR. Scale bar = 20 μm. ( B ): The average radial distance of the longest neurite, measured from the neurite tip to the soma, of 50 cells was determined after 24 h treatment with test drugs. Results represent the mean ± SEM. Significant difference in values between the compound-treated and non-treated cells: ** P

    Journal: International Journal of Molecular Sciences

    Article Title: Neurotrophic Effect of Citrus Auraptene: Neuritogenic Activity in PC12 Cells

    doi: 10.3390/ijms13055338

    Figure Lengend Snippet: Effects of AUR on neurite outgrowth from PC12 cells. Cells were treated with various concentrations (0, 10, 30, and 50 μM) of AUR or NGF (50 ng/mL) for 24 h, after which phase-contrast photomicrographs of the cells were taken ( A ): a , control; b , treatment with NGF; c , treatment with 30 μM AUR. Scale bar = 20 μm. ( B ): The average radial distance of the longest neurite, measured from the neurite tip to the soma, of 50 cells was determined after 24 h treatment with test drugs. Results represent the mean ± SEM. Significant difference in values between the compound-treated and non-treated cells: ** P

    Article Snippet: Mouse NGF 2.5S (Grade I) was purchased from Alomone Labs Ltd. (Jerusalem, Israel).

    Techniques: