Article Figures & Data
Figures
- Fig. 1.
Immunoblot analysis of MAP2 isoforms present in adult rat olfactory bulb. Equal aliquots of total bulb homogenate were separated on 8% gels, transferred to nitrocellulose, and probed with the indicated antibodies against MAP2. Lane 1, Antibody HM-2, which recognizes all known isoforms of MAP2 independent of phosphorylation state; lane 2, antibody #266, which recognizes only high molecular weight isoforms of MAP2 independent of phosphorylation state; lane 3, antibody AP18, which recognizes all known isoforms of MAP2 only when phosphorylated on Ser136. Positions of high molecular weight MAP2 and low molecular weight MAP2c are indicated by arrows. The fainter bands running below MAP2 and MAP2c likely represent proteolytic breakdown products. Positions of molecular weight markers (in kilodaltons) are given on the right.
- Fig. 2.
Photomicrograph depicting immunoreactivity for a phosphorylation-independent MAP2 antibody, antibody 266, in a coronal section through the olfactory bulb of a P60 rat. MAP2-IR is absent in the olfactory nerve layer (ONL) and subependymal zone (SUB). MAP2-IR dendrites have extensive ramifications in neuropil of the glomerular layer (GLM). The external plexiform layer (EPL) is dense with MAP2-IR dendrites. Numerous MAP2-IR dendrites extend from the granule cell layer (GCL), through the mitral cell layer (MCL), and into theEPL. Scale bar, 300 μm.
- Fig. 3.
Photomicrographs depicting immunoreactivity for a phosphorylation-independent MAP2 antibody, antibody #266, in coronal sections of the developing bulb. A, At P10, MAP2-IR dendrites have extensive arborizations in glomeruli, and the external plexiform layer contains numerous MAP2-IR dendrites. Granule cells have distinct MAP2-IR along their somatic periphery, and their dendrites are lightly labeled. B, At P20, MAP2-IR is most intense at this age. The deep half of the external plexiform layer generally has more labeling than does the superficial half. Granule cell dendrites are darkly immunoreactive. C, At P30, MAP2-IR has a distribution similar to that seen in the P20 bulb. D, At P60, the intensity of MAP2-IR has decreased slightly from that observed in the P30 bulb. Arrows mark the mitral cell layer. For abbreviations, see Figure 2. Scale bar, 200 μm.
- Fig. 4.
Graph depicting the mean percent difference (±SEM) between left and right bulbs in the area fraction of MAP2-IR (phosphorylation-independent antibody 266) within the granule cell layer. In sham-operated animals (SHAM), there is no difference between bulbs in MAP2-IR from P10, P20, P30, or P60 rats. In naris-occluded rats (NOSX), there is also a negligible difference in MAP2-IR between bulbs from animals occluded from P1 to P10, P1 to P20, P1 to P30, or P30 to P60.
- Fig. 5.
Photomicrographs depicting immunoreactivity for a phosphorylation-independent antibody to MAP2 (A), antibody 266, and staining for the phosphoepitope-specific AP18 antiserum (B) in adjacent coronal sections of bulbs from a P30 rat. A, Numerous dendrites in the glomerular layer contain MAP2-IR. Labeling of dendrites is particularly dense along the deep border of the glomerular neuropil. The external plexiform layer has dark immunoreactivity, and stained dendrites course from the external plexiform layer through the mitral cell layer.B, AP18-IR is generally much sparser than is staining for the phosphorylation-independent MAP2 antibody. For example, relatively few dendrites are labeled in glomeruli. Labeling is less dense in the external plexiform and granule cell layers as well. Thearrow marks the mitral cell layer. For abbreviations, see Figure 2. Scale bar, 100 μm.
- Fig. 6.
Photomicrographs demonstrating AP18-IR in coronal sections from control (left) and experimental (right) bulbs after unilateral naris closure.Left, AP18-IR dendrites are apparent as they project into glomeruli or extend laterally in superficial aspects of the external plexiform layer. AP18-IR is relatively dense in the external plexiform layer, and many dendrites in the granule cell layer are labeled. Right, AP18-IR is less intense in experimental bulbs. Fewer dendrites are labeled in the glomerular layer, and there is substantially less immunoreactivity of granule cell dendrites. Thearrow marks the mitral cell layer. For abbreviations, see Figure 2. Scale bar, 100 μm.
- Fig. 7.
Graph of the mean percent difference (± SEM) between left and right bulbs in the area fraction of AP18-IR within the granule cell layer. In sham-operated rats (SHAM), there is a negligible difference between right and left bulbs in P10, P20, P30, and P60 rats. However, after naris closure (NOSX), the area fraction of the bulb containing AP18-IR in the granule cell layer is dramatically reduced in experimental bulbs after occlusion from P1 to P20, P1 to P30, or P30 to P60. Asterisks denote significant differences from zero (p < 0.05). Note the y-axis change from Figure 4.
- Fig. 8.
Photomicrograph depicting HM-2 staining in a coronal section from a P30 bulb. The HM-2 antibody is phosphorylation-independent and recognizes all known forms of MAP2. Note the absence of staining in the nerve layer and the particularly dense immunoreactivity in the deeper portion of the external plexiform layer. Staining patterns are similar to those found using antibody 266. For abbreviations, see Figure 2. Scale bar, 150 μm.
- Fig. 9.
Photomicrograph depicting calcineurin-IR in a coronal section through the granule cell layer of a P20 bulb. Calcineurin-IR is located within somata but absent from the nucleus. There is some faint labeling of dendrites. For abbreviations, see Figure 2. Scale bar, 75 μm.
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