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1.
Figure 2

Figure 2. ID elements do not mark EGFP reporter mRNA for dendritic transport.. From: Can ID Repetitive Elements Serve as Cis-acting Dendritic Targeting Elements? An In Vivo Study.

In situ hybridization using a digoxigenin-labeled antisense (AS) EGFP probe on coronal brain sections from the various transgenic mice. A–D, There are intense hybridization signals in the cell bodies of neurons in the hippocampus and cortex for the ID1, ID2, ID4 and 1DI chimeric RNAs; however, there is no staining observed in the corresponding dendritic fields. E–H, Higher magnifications of the CA1 regions of hippocampus reveal intense staining in the cell bodies (arrowheads) but only weak staining, if at all, in only the very proximal-most regions of dendrites (arrows) and none in the distal dendritic fields (asterisks). I, An example of dendritic staining; brain sections from wild type mice were hybridized with a unique probe for BC1 RNA. J, Control hybridizations with the EGFP sense (S) probe did not produce any signals. Boxes in A–D represent the areas shown at higher magnification in E–H, respectively.

Tasneem Khanam, et al. PLoS ONE. 2007;2(9):e961.
2.
Figure 1

Figure 1. Schematic representation of transgenic transcripts and chimeric constructs containing EGFP plus various cis elements.. From: Can ID Repetitive Elements Serve as Cis-acting Dendritic Targeting Elements? An In Vivo Study.

The cis elements (ID1, ID2, ID4, 1DI) were inserted downstream (within the 3′ UTR) of the EGFP cDNA, and followed by polyadenylation signals from SV40 (spotted box). The chicken β-actin promoter was used to drive transcription of the chimeric mRNAs. The promoter harbors an intron of about 900 bp (hatched box). The various ID elements are represented by black rectangular boxes, with short A-rich tails (white boxes). ID1 corresponds to the 5′domain of BC1 RNA and the other ID elements differ from ID1 by mutations indicated by white lines within the black boxes, 1DI corresponds to ID1 in the reverse orientation. BC1-FL (full length BC1 RNA) is indicated by a black rectangular box (5′ ID domain) fused to two open boxes (representing the central A-rich region and 3′ non-repetitive region). The 3′ UTR of α-CaMKII mRNA, used as a positive control, is represented by a long rectangular open box and part of the ORF by a striped box (not drawn to scale). The labels to the right refer the respective plasmids.

Tasneem Khanam, et al. PLoS ONE. 2007;2(9):e961.
3.
Figure 3

Figure 3. The 3′ UTR of α-CaMKII targets EGFP reporter mRNA to dendrites, whereas full-length BC1 RNA does not.. From: Can ID Repetitive Elements Serve as Cis-acting Dendritic Targeting Elements? An In Vivo Study.

In situ hybridization with a DIG-riboprobe against EGFP in coronal brain sections through the hippocampus of transgenic mice harboring chimeric RNAs with EGFP plus full length BC1 RNA or the 3′ UTR of α-CaMKII mRNA (positive control). A, EGFP-BC1 RNA was restricted to neuronal cell bodies in the hippocampus and cortex (arrowheads), and no significant labeling was observed in the dendritic regions of these cells. B, Higher magnification revealed dendritic labeling only in the very proximal-most region of the CA1 pyramidal cell dendrites (arrows), if at all. C, The EGFP- α-CaMKII 3′ UTR chimeric RNA was detected both in cell bodies and dendrites of neurons in the hippocampus and cortex. D, Higher magnification revealed somatic staining (arrowheads) and labeling up to the distal ends of CA1 pyramidal cell dendrites in the hippocampus (arrows). E-F, There were no specific hybridization signals from AS probes in wild type mice, or from sense probes in the transgenic mice (data not shown). Boxes in A and C represent the areas shown at higher magnification in B and D, respectively.

Tasneem Khanam, et al. PLoS ONE. 2007;2(9):e961.

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