Local translation of classes of mRNAs that are targeted to neuronal dendrites - PubMed (original) (raw)

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Local translation of classes of mRNAs that are targeted to neuronal dendrites

J Eberwine et al. Proc Natl Acad Sci U S A. 2001.

Abstract

The functioning of the neuronal dendrite results from a variety of biological processes including mRNA transport to and protein translation in the dendrite. The complexity of the mRNA population in dendrites suggests that specific biological processes are modulated through the regulation of dendritic biology. There are various classes of mRNAs in dendrites whose translation modulates the ability of the dendrite to receive and integrate presynaptic information. Among these mRNAs are those encoding selective transcription factors that function in the neuronal soma and ionotropic glutamate receptors that function on the neuronal membrane. Conclusive evidence that these mRNAs can be translated is reviewed, and identification of the endogenous sites of translation in living dendrites is presented. These data, as well as those described in the other articles resulting from this colloquium, highlight the complexity of dendritic molecular biology and the exquisitely selective and sensitive modulatory role played by the dendrite in facilitating intracellular and intercellular communication.

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Figures

Figure 1

Figure 1

Differential display analysis of dendritic processes from a single hippocampal neuron in culture. (A) Isolated hippocampal cells free of overlapping processes from neighboring cells were identified in low-density cultures (5). Individual dendrites were harvested by transecting at the branch point and aspirated as described (3). (B) Comparisons of differential display products between dendrites with a single 5′ primer, OPA-5 (Operon Technologies, Alameda, CA), in combination with anchor primers A (T11AC) or C (T11GC) show the presence of common (closed arrowheads) and unique (open arrowheads) PCR products. (C) When a single 5′ primer (OPA-13) is used in combination with all nine anchor primers a large population of mRNAs are present within neuronal processes.

Figure 2

Figure 2

Time course for movement of mRNAs into the neuronal dendrite. Intact cultured rat hippocampal neurons were treated with DHPG. mRNA was harvested from dendrites at times of 0, 24, 32, and 45 min posttreatment, and aRNA was amplified. This probe was used to screen various types of arrays including the macroarrays shown. The hybridization intensity of selected spots corresponding to different immobilized cDNAs increased as a function of time after DHPG treatment. These data indicate that the movement of mRNA into the dendrite can be regulated by DHPG.

Figure 3

Figure 3

Transfection of isolated dendrites with GFP mRNA results in fluorescence. (A and B) Phase images of 4-day-old primary hippocampal neurons grown on grided coverslips. Arrows indicate positions of cell bodies removed with a micropipette. (C) Two identical enlarged images of one dendrite from B. Bold black line indicates the position of the dendrite. (D) Four fluorescence images of dendrite in B before (0) and after (63, 133, and 203 min) transfection with GFP mRNA. Images were contrast-stretched in National Institutes of Health

image

for display purposes. (Scale bars = 50 μm.) (E) Mean fluorescence in isolated dendrites over a time course of hours. Black bar indicates period of transfection with GFP mRNA. n = 3.

Figure 4

Figure 4

Stimulation of glutamate receptor mRNA translation in isolated dendrites. (A and C) The phase-contrast images of transected dendrites that correspond to dendrites that have been transfected with GluR2-c-myc shown in B and D, respectively. The dendrites in D have been treated with DHPG to stimulate protein synthesis, whereas those in B are DHPG naïve. Arrows point to individual dendrites that are visible in the paired panels. See ref. for more information.

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