Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: A. Representative micrographs of TH immunohistochemistry. Headings = Anterior-posterior coordinates of Paxinos and Watson Rat Atlas . 24WLUT = Untransfected rats with 24 weeks of 6-OHDA lesion. 12WL4WT = 12 weeks after lesion and 4 weeks after transfection, 12WL12WT = 12 weeks after lesion and 12 weeks after transfection. The scale bar of 300 μm is common for slices at different levels and of 50 μm for details. B. TH(+) neuron counting. C. Densitometry of TH(+) relative area. 4WT and 12 WT = 4 and 12 weeks after transfection. 24WL = untransfected rats with 24 weeks of lesion. 12WL = 12 weeks after lesion. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion. All values are the mean ± SEM ( n = 3 independent rats for each experimental condition). One-way ANOVA and Newman-Keuls post-test. * P < 0.001, ** P < 0.0001. ns = no statistical significance, P > 0.05.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Immunohistochemistry, Transfection

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: A. Representative micrographs of TH immunohistochemistry. Headings = Anterior-posterior coordinates of Paxinos and Watson Rat Atlas . 24WLUT = Untransfected rats with 24 weeks of 6-OHDA lesion. 12WL4WT = 12 weeks after lesion and 4 weeks after transfection, 12WL12WT = 12 weeks after lesion and 12 weeks after transfection. The scale bar of 1 mm is for slices at different levels and of 100 μm for details. B. Densitometry of TH(+) relative area. 4WT and 12 WT = 4 and 12 weeks after transfection. 24WL = untransfected rats with 24 weeks of lesion. 12WL = 12 weeks after lesion. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion. One-way ANOVA and Newman-Keuls post-test. * P < 0.001, ** P < 0.0001. ns = no statistical significance, P > 0.05.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Immunohistochemistry, Transfection

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: A. Representative micrographs of double immunofluorescence staining against NTSR1 and TH. 24WLUT = Untransfected rats with 24 weeks of 6-OHDA lesion. 12WL4WT = 12 weeks after lesion and 4 weeks after transfection, 12WL12WT = 12 weeks after lesion and 12 weeks after transfection. The scale is common for all micrographs. B. immunofluorescence area density (IFAD) for NTSR1 and TH was determined using ImageJ software v.1.46r (National Institutes of Health; Bethesda, MD). NRTN4WT and NRTN412WT = 4 and 12 weeks after transfection. UT = untransfected rats. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion. All values are the mean ± SEM ( n = 3 independent rats for each experimental condition). Two-way ANOVA and Bonferroni post-test. ns = no statistical significance.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Double Immunofluorescence Staining, Transfection, Immunofluorescence, Software

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: A. Representative micrographs of double immunofluorescence staining against NTSR1 and TH. 24WLUT = Untransfected rats with 24 weeks of 6-OHDA lesion. 12WL4WT = 12 weeks after lesion and 4 weeks after transfection, 12WL12WT = 12 weeks after lesion and 12 weeks after transfection. The scale is common for all micrographs. B. immunofluorescence area density (IFAD) for NTSR1 and TH was determined using ImageJ software v.1.46r (National Institutes of Health; Bethesda, MD). NRTN4WT and NRTN412WT = 4 and 12 weeks after transfection. UT = untransfected rats. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion. All values are the mean ± SEM (n = 3 independent rats for each experimental condition). Two-way ANOVA and Bonferroni post-test. ns = no statistical significance.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Double Immunofluorescence Staining, Transfection, Immunofluorescence, Software

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: Representative micrographs of double immunofluorescence to TH and β-III-tubulin. 24WLUT = Untransfected rats with 24 weeks of 6-OHDA lesion. 12WL4WT = 12 weeks after lesion and 4 weeks after transfection, 12WL12WT = 12 weeks after lesion and 12 weeks after transfection. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion. The scale bar is common for all micrographs.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Immunofluorescence, Transfection

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: A . Sholl diagram for the morphological study of dendrites and dendritic spines. B . Representative micrograph of a medium spiny neuron (MSN) of a healthy striatum. The arrow shows the 50 μm segment (primary dendrite) where dendritic spines were analyzed. Upper calibration bar = 25 μm. The small panels show different types of spines (thin, mushroom, stubby and wide, from left to right) indicated by the arrow. Calibration bar = 5 μm. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion and the Sholl analysis was performed at the end of the study (12 weeks after transfection or 24 weeks after lesion). UT = untransfected rats with lesion. C. Sholl analysis of total number of intersections along dendritic trees in MSNs at all distances in 200-μm radius from the soma. D. Plot of mean spine density analyzed per 50-μm primary dendrite from cell body (proximal segment, upper arrow in B ) from six neurons per rat. All values represent the mean ± SEM ( n = 6 independent rats in each experimental condition). Statistical analysis ( C and D ) was performed using one-way ANOVA and Tukey post-hoc test.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Transfection

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: Ipsilateral circling behavior ( A and C ) was activated by methamphetamine (8 mg/kg; i.p.), and contralateral circling behavior ( B and D ) was activated by apomorphine (0.5 mg/kg; i.p.) at weeks 0, 4 and 12 after transfection. The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion (Week 0). All values represent the mean ± SEM ( n = 4 independent rats in each experimental condition). One-way ANOVA and Newman-Keuls post-test. * P <0.0005, ** P < 0.0001.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Transfection

Journal: PLoS ONE

Article Title: Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion

doi: 10.1371/journal.pone.0188239

Figure Lengend Snippet: The transfections of pGreenLantern-1 (GFP) and pTracer-mNRTN-His (NRTN) plasmids were made at week 12 after lesion. UT = untransfected rats. The tests were performed at week 4 (12WL4WT) and week 12 (12WL12WT) after transfection. All values represent the mean ± SEM ( n = 4 independent rats in each experimental condition). * P < 0.001 when compared the values of the contralateral limb (Contra limb) with those of the ipsilateral limb (Ipsi limb), ** P < 0.001 when compared NRTN-His vs UT and § P < 0.001, NRTN-His vs GFP using two-way ANOVA. The post-hoc test was Newman-Keuls analysis.

Article Snippet: Full NRTN sequence and correct Tag addition were confirmed by automatic sequencing (BigDye v3.1; Applied Biosystems; Foster City, CA, USA). pGreenLantern-1 (5,030 bp) coding for GFP under the control of CMV promoter (Gibco BRL; Grand Island, NY, USA) was used as a negative control.

Techniques: Transfection