Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Immunohistochemical distribution pattern of MMP 2 (A), MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and TIMP 3 (F) in the cerebellar cortex of 10-d-old rats. Immunohistochemical labeling was performed on 10-μm-thick frontal rat cerebellar sections showing the EGL, the ML, the PCL, and the IGL. The labeling with polyclonal antibodies was enhanced using the fluorescein tyramide signal amplification system (New England Nuclear). In the EGL, MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and to less extents for TIMP 3 (F), were seen in granular precursor cells. In addition, MMP 9 was detected along the Bergmann glial fibers in this layer (C).Arrows point to strips of stained granular precursor cells. In the ML, MMP 3, MMP 9, and TIMP 1 were diffusely present over PK cells dendrites, whereas TIMP 3 was restricted to the cytoplasm of the PK cell dendrites (the small arrow inF points to the labeled PK cell dendritic arborization). In the PCL, the PK cell somata showed labeling for each marker, and MMP 9 expression was seen in the Bergmann glial cell somata. Some granular cells in the IGL were immunopositive for MMPs 3 and 9 and TIMPs 1, 2, and 3. Scale bar, 50 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Immunohistochemical staining, Labeling, Amplification, Staining, Marker, Expressing

Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Immunohistochemical distribution pattern of MMP 2 (A), MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and TIMP 3 (F) in the cerebellar cortex of 15-d-old rats. Ten-micrometer-thick frontal sections of rat cerebellum showed the EGL, the ML, the PCL, and the IGL. Immunohistochemical staining was performed using polyclonal antibodies and amplified by tyramide signal amplification (New England Nuclear). EGL immunostaining for MMPs 3 and 9 and TIMPs 1, 2, and 3 was lost. The PK cell dendrites were still stained for MMP 3 and TIMP 3. Note the diffuse extracellular distribution of MMP 3 as opposed to the cytoplasmic distribution of TIMP 3. ML labeling for MMP 9 and TIMP 1 in the PK cell dendrites decreased, but MMP 9 labeling of Bergmann glial fibers was maintained. MMP 9 labeling in ML cells, probably corresponding to stellate and basket cells, was detected. Thelarge double arrows point to MMP 9-positive Bergmann glial fibers, and the small arrows point to MMP 9-positive ML cells (C). In the PCL, expression of MMPs 2, 3, and 9 and TIMPs 1, 2, and 3 was noted in the PK cell somata. TIMP 1 was differentially expressed in clusters of PK cell somata (stained and unstained PK cell somata are respectively indicated by large arrows at the left andright of D). TIMP 1 was also present in Bergmann glial somata around labeled and unlabeled PK cell bodies (D). Granular cells in the IGL were positive for MMPs 3 and 9 and TIMPs 1 and 3. Scale bar, 25 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Immunohistochemical staining, Staining, Amplification, Immunostaining, Labeling, Expressing

Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Immunohistochemical distribution pattern of MMP 2 (A), MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and TIMP 3 (F) in adult rat cerebellar cortex. Ten micrometer frontal sections of adult rat cerebellum showed the EGL, the ML, the PCL, and the IGL. Labeling with polyclonal antibodies was amplified by using tyramide signal amplification (New England Nuclear). Note, for each marker, the cytoplasmic staining at the adult age compared with the diffuse staining seen on the developmental stages P10 and P15. In the ML, MMPs 3 and 9 and TIMPs 1, 2, and 3 protein expression was lost in PK cell arborization. However, MMP 9 expression persisted in the Bergmann glial fibers. Double arrows point to one fiber in C. Only the anti-TIMP 3 antibodies labeled PK cells dendrites. Arrow indicates TIMP 3 dendritic immunolabeling in F. Interneurons of the ML were labeled for MMPs 3 and 9 and TIMPs 1 and 2. In the PCL, all markers were detected in PK cell somata. In addition, MMP 9 and TIMP 1 were seen in the Bergmann glial cell bodies surrounding the PK cell somata. In the IGL, MMPs 3 and 9 and TIMPs 2 and 3 were strongly expressed in many granular cells, whereas TIMP 1 was poorly expressed. Magnification of the left of each panel, 100×. Magnification of theright of each panel, 400×. Scale bars, 100 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Immunohistochemical staining, Labeling, Amplification, Marker, Staining, Expressing, Immunolabeling

Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Immunohistochemical distribution pattern of MMP 2 (A), MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and TIMP 3 (F) in the cerebellar cortex of 10-d-old rats. Immunohistochemical labeling was performed on 10-μm-thick frontal rat cerebellar sections showing the EGL, the ML, the PCL, and the IGL. The labeling with polyclonal antibodies was enhanced using the fluorescein tyramide signal amplification system (New England Nuclear). In the EGL, MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and to less extents for TIMP 3 (F), were seen in granular precursor cells. In addition, MMP 9 was detected along the Bergmann glial fibers in this layer (C).Arrows point to strips of stained granular precursor cells. In the ML, MMP 3, MMP 9, and TIMP 1 were diffusely present over PK cells dendrites, whereas TIMP 3 was restricted to the cytoplasm of the PK cell dendrites (the small arrow inF points to the labeled PK cell dendritic arborization). In the PCL, the PK cell somata showed labeling for each marker, and MMP 9 expression was seen in the Bergmann glial cell somata. Some granular cells in the IGL were immunopositive for MMPs 3 and 9 and TIMPs 1, 2, and 3. Scale bar, 50 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Immunohistochemical staining, Labeling, Amplification, Staining, Marker, Expressing

Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Immunohistochemical distribution pattern of MMP 2 (A), MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and TIMP 3 (F) in the cerebellar cortex of 15-d-old rats. Ten-micrometer-thick frontal sections of rat cerebellum showed the EGL, the ML, the PCL, and the IGL. Immunohistochemical staining was performed using polyclonal antibodies and amplified by tyramide signal amplification (New England Nuclear). EGL immunostaining for MMPs 3 and 9 and TIMPs 1, 2, and 3 was lost. The PK cell dendrites were still stained for MMP 3 and TIMP 3. Note the diffuse extracellular distribution of MMP 3 as opposed to the cytoplasmic distribution of TIMP 3. ML labeling for MMP 9 and TIMP 1 in the PK cell dendrites decreased, but MMP 9 labeling of Bergmann glial fibers was maintained. MMP 9 labeling in ML cells, probably corresponding to stellate and basket cells, was detected. Thelarge double arrows point to MMP 9-positive Bergmann glial fibers, and the small arrows point to MMP 9-positive ML cells (C). In the PCL, expression of MMPs 2, 3, and 9 and TIMPs 1, 2, and 3 was noted in the PK cell somata. TIMP 1 was differentially expressed in clusters of PK cell somata (stained and unstained PK cell somata are respectively indicated by large arrows at the left andright of D). TIMP 1 was also present in Bergmann glial somata around labeled and unlabeled PK cell bodies (D). Granular cells in the IGL were positive for MMPs 3 and 9 and TIMPs 1 and 3. Scale bar, 25 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Immunohistochemical staining, Staining, Amplification, Immunostaining, Labeling, Expressing

Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Immunohistochemical distribution pattern of MMP 2 (A), MMP 3 (B), MMP 9 (C), TIMP 1 (D), TIMP 2 (E), and TIMP 3 (F) in adult rat cerebellar cortex. Ten micrometer frontal sections of adult rat cerebellum showed the EGL, the ML, the PCL, and the IGL. Labeling with polyclonal antibodies was amplified by using tyramide signal amplification (New England Nuclear). Note, for each marker, the cytoplasmic staining at the adult age compared with the diffuse staining seen on the developmental stages P10 and P15. In the ML, MMPs 3 and 9 and TIMPs 1, 2, and 3 protein expression was lost in PK cell arborization. However, MMP 9 expression persisted in the Bergmann glial fibers. Double arrows point to one fiber in C. Only the anti-TIMP 3 antibodies labeled PK cells dendrites. Arrow indicates TIMP 3 dendritic immunolabeling in F. Interneurons of the ML were labeled for MMPs 3 and 9 and TIMPs 1 and 2. In the PCL, all markers were detected in PK cell somata. In addition, MMP 9 and TIMP 1 were seen in the Bergmann glial cell bodies surrounding the PK cell somata. In the IGL, MMPs 3 and 9 and TIMPs 2 and 3 were strongly expressed in many granular cells, whereas TIMP 1 was poorly expressed. Magnification of the left of each panel, 100×. Magnification of theright of each panel, 400×. Scale bars, 100 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Immunohistochemical staining, Labeling, Amplification, Marker, Staining, Expressing, Immunolabeling

Journal: The Journal of Neuroscience

Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum

doi: 10.1523/JNEUROSCI.19-12-04994.1999

Figure Lengend Snippet: Differential distribution of MMP 9 (A–C), TIMP 2 (D–F), and TIMP 3 (G–I) transcripts in the developing and adult rat cerebellum. In situhybridization was performed on frontal rat cerebellum sections (12 μm) using antisense MMP 9, TIMP 2, and TIMP 3 mRNA oligoprobes provided by Biognostik. Autoradiographic film images (A,B, D, E, G,H) were obtained by exposure for 5 d on Hyperfilm-βmax film (Amersham). The exposure time for the emulsion-coated sections (C, F,I) was 6 weeks. On P10, MMP 9 mRNA was intensely expressed in the whole EGL and weakly expressed in the ML, PCL, and IGL (A). In the adult, the global labeling was restricted to the PCL and IGL (B). MMP 9 mRNA subcellular expression was detected in ML interneurons (small arrows) and PK cell somata (large arrows) (C). TIMP 2 mRNA expression was mainly observed in the EGL on P10 (D) and in the PCL and IGL in the adult (E). On emulsion-coated sections, TIMP 2 was visible in the ML interneurons (small arrows), in the border of PK cell somata, and in the IGL (F). On P10, TIMP3 transcript expression was high in the EGL (G). No expression of TIMP 3 was seen in the ML on autoradiographic films, but labeling for TIMP 3 was seen in the PCL and IGL of the 10 d cerebellum (G). In the adult, labeling was faint in the ML and high in the PCL and IGL (H). TIMP 3 mRNA was seen in the ML interneurons (small arrows) and in some PK cell somata (large arrow) (I).WM, White matter. Scale bars: A,D, G, 1 and 3 mm;C, F, I, 40 μm.

Article Snippet: The primary polyclonal rabbit antibodies, used at a 1:2500 dilution, were anti-MMP 2 (hinge region), anti-MMP 3 (hinge region), anti-MMP 9 (N-terminal region of the active form), anti-TIMP 1 (C-terminal region), anti-TIMP 2, and anti-TIMP 3 (Chemicon, Temecula, CA).

Techniques: Labeling, Expressing