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4 publications mentioning dre-mir-153c

Open access articles that are associated with the species Danio rerio and mention the gene name mir-153c. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

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[+] score: 371
miR-153 Targets snap-25 To identify mRNAs regulated by miR-153, we used target prediction algorithms, compared the expression patterns of both potential mRNA targets and miR-153, and assayed phenotypes from gain and loss of function experiments. [score:8]
Although miR-153 is likely to have additional targets, the ability to specifically rescue the effects of overexpression and knockdown of both miR-153 and snap-25 indicates that the effects we observe are specific to targeting of snap-25 by miR-153. [score:8]
Error bars show s. e. m. miR-153 Regulates snap-25 to Control MovementBecause we could specifically suppress the effects of overexpression or knockdown of miR-153 by co-injection of either snap-25a,b mRNA or morpholinos against snap-25a,b, we next sought to test whether the movement defects are caused by altered miR-153 levels could likewise be rescued in a snap-25 dependent manner. [score:7]
To identify mRNAs regulated by miR-153, we used target prediction algorithms, compared the expression patterns of both potential mRNA targets and miR-153, and assayed phenotypes from gain and loss of function experiments. [score:6]
Error bars show s. e. m. Because we could specifically suppress the effects of overexpression or knockdown of miR-153 by co-injection of either snap-25a,b mRNA or morpholinos against snap-25a,b, we next sought to test whether the movement defects are caused by altered miR-153 levels could likewise be rescued in a snap-25 dependent manner. [score:6]
Overexpression of miR-153 caused decreased FM1-43 loading, indicating down-regulation of the synaptic vesicle cycle within NMJ boutons (arrowheads). [score:6]
The levels of GFP mirrored the effects observed using fluorescence imaging in live embryos–reduced reporter expression in the presence of miR-153 and increased reporter expression upon knockdown of miR-153 (Fig. 2C,D). [score:6]
Nevertheless, the effects in this case were fully suppressed by co -expression of either miR-153/snap-25a,b mRNA or MOs against miR-153/snap-25a,b, demonstrating specific regulation of snap-25 by miR-153. [score:6]
An intriguing possibility based on the results presented here is that developmental, stage-specific and/or cell-specific expression of miR-153 may similarly regulate SNAP-25 levels, which then drives developmental and cell-specific effects. [score:6]
Expression of miR-153 in Motor NeuronsTo ensure that the effects of miR-153 on motor neuron patterning were due to expression of miR-153 in these cells, we FACS sorted cells from the trunks of 52 hpf (Tg(mnx1:TagRFP-T) embryos and conducted RT/qPCR. [score:5]
Prior work had shown that miR-153 is expressed in the brain and spinal cord but these results show that miR-153 is expressed in developing motor neurons. [score:5]
Using deep sequencing and in situ localization, we detected robust miR-153 expression in the developing zebrafish brain and reduced, but detectable levels in the spinal cord as early as the 18 somite stage, with progressively increasing expression thereafter [30], [31] [32]. [score:5]
In this study, we show that miR-153 inhibits SNAP-25 expression in the developing nervous system. [score:5]
Overexpression of miR-153 or knockdown of snap-25a,b (snap-25a,b [MO]) caused severe defects in axon development and architecture (asterisks). [score:5]
miR-153 also likely targets other mRNAs [80], but SNAP-25 regulation alone is required and sufficient to explain the role of miR-153 regulation of movement, motor neuron morphogenesis, and SNARE -mediated secretion. [score:5]
miR-153 regulates endogenous snap-25a expression. [score:4]
Overexpression of miR-153 and knockdown of snap-25a,b (snap-25a,b [MO]) reduced the levels of hGH to below the amount detected in culture media from mock transfected cells (Fig. 9). [score:4]
Figure S1 Northern blot of miR-153 overexpression and knockdown. [score:4]
At 52 hpf, miR-153 overexpression fish embryos were still mostly motionless, while miR-153 knockdown embryos were still hyperactive (data not shown). [score:4]
If miR-153 targets snap-25, knockdown of endogenous miR-153 should lead to increased reporter fluorescence. [score:4]
In sharp contrast, knockdown of miR-153 and overexpression of snap-25 both significantly increased the amount of secreted hGH 8–10 fold over the mock transfected control (Fig. 9). [score:4]
0057080.g003 Figure 3 miR-153 regulates endogenous snap-25a expression. [score:4]
For secondary motor neurons, rostral axon outgrowth was similarly stunted and/or irregularly spaced by miR-153 overexpression and slightly elongated by miR-153 knockdown (Fig. S6). [score:4]
Based on fluorescence levels in live embryos at 1 dpf, co-injection of miR-153 resulted in obvious down-regulation of GFP for both isoforms (Fig. 2B). [score:4]
Knockdown of snap-25 resulted in dramatically decreased embryonic movements, similar to overexpression of miR-153 (Fig. 1). [score:4]
One hour later, western blots were performed on pooled protein samples to determine whether it was possible to rescue SNAP-25 over -expression phenotypes associated with miR-153 knockdown or injection of snap25a,b mRNAs. [score:4]
Regulated expression of miR-153 provides an attractive mo del to mechanistically explain tight control of SNAP-25 levels. [score:4]
Arrowheads indicate the structural defects after miR-153 overexpression or knockdown of snap-25a,b (snap-25a,b [MO]). [score:4]
The significant difference between miR-153 knockdown and overexpression conditions indicates that miR-153 plays an important role in controlling the rate of vesicle cycling (Fig. 8D). [score:4]
Together, these experiments strongly support the conclusion that miR-153 specifically targets snap-25 to regulate embryonic movement. [score:4]
Injection of snap-25a,b mRNA or morpholinos against snap-25a/b produced virtually the same phenotypes observed in embryos subjected to miR-153 knockdown or overexpression, respectively. [score:4]
Arrows indicate increased branching after knockdown miR-153 (miR-153 [MO]) or overexpression snap-25a,b mRNA. [score:4]
Focusing on rostral effects, injection of snap-25a,b mRNA phenocopied miR-153 knockdown and injection of morpholinos against snap-25 resulted in patterns that closely resembled miR-153 overexpression. [score:4]
Conversely, miR-153 knockdown causes elevated SNAP-25 expression resulting in hyperactive movement, increased neuronal secretion, and increased neuronal growth/branching. [score:4]
Exposure to BoNT A dramatically reduced SNAP-25 levels, recapitulating the effects of miR-153 knockdown and over -expression (Fig. 4A,B). [score:4]
Two different morpholinos were used to ensure specificity and we verified overexpression and knockdown of miR-153 using northern blots (Fig. S1). [score:4]
In this study, we show that miR-153 regulates the critical core SNARE component, SNAP-25, to modulate exocytosis and neuronal development. [score:3]
These results indicate that miR-153 regulates motor neuron development via control of snap-25a,b. [score:3]
Effects of miR-153 Overexpression on Movement at 24 hpf Single cell zebrafish embryos were injected with miR-153 and filmed for one minute at 24 hpf. [score:3]
miR-153 regulates primary motor neuron development. [score:3]
miR-153 Targets snap-25. [score:3]
Based on these criteria, snap-25 proved to be a bona fide target for miR-153 based on the results of reporter silencing experiments (Fig. 2) and consistent with conservation of miRNA recognition elements (MREs) from fish to humans (Fig. S2). [score:3]
Increased miR-153 levels cause decreased SNAP-25 expression resulting in decreased embryonic movement, decreased neuronal secretion, and decreased neuronal growth/branching. [score:3]
We conclude that miR-153 targets both isoforms of snap-25 in an MRE -dependent manner. [score:3]
Figure S5 Dose -dependent rescue of miR-153 over -expression. [score:3]
This may indicate a possible additional function for miR-153 in regulating axonal growth and patterning during secondary motor neuron development. [score:3]
We next tested whether miR-153 targets endogenous snap-25. [score:3]
To test whether miR-153 plays a role in this secretory context, we examined exocytosis in a rat neuroendocrine pituitary cell line (GH4C1) expressing human growth hormone (hGH) [51]. [score:3]
Figure S6 miR-153 regulates secondary motor neuron development. [score:3]
miR-153 is expressed in motor neurons. [score:3]
We propose that miR-153 control of SNAP-25 levels allows for precise regulation of SNAP-25 during development and exocytosis. [score:3]
Our work demonstrates that miR-153 is a member of this subset of miRNAs implicated in neuronal function but by a distinctly different mechanism through targeting of snap-25. [score:3]
GH4C1 cells were therefore transfected with miR-153, morpholinos against miR-153/ snap-25, or vectors expressing snap-25a,b, followed by determination of hGH levels in the media by ELISA. [score:3]
Perturbation of miR-153 expression levels by injection of miR-153 or MOs against different regions of pre- miR-153 was verified by northern blot. [score:3]
Thus, not only were SNAP-25 protein levels restored to normal, but also movement defects were rescued, demonstrating specific targeting of snap-25 by miR-153. [score:3]
If miR-153 is targeting snap-25, the effects of increased miR-153 should mimic the effects of BoNT A. To test this prediction, injected zebrafish were exposed to BoNT A for 30 minutes at 27 hpf. [score:3]
Compared to NIC embryos, a striking difference in primary motor neuron axon architecture was observed with both miR-153 overexpression (miR-153) and knockdown (miR-153 [MO])(Fig. 6). [score:3]
Cleavage of SNAP-25 by Botulinum neurotoxin A causes a paralytic phenotype that resembles the loss of movement we observe in zebrafish embryos expressing excess miR-153. [score:3]
miR-153 targets snap-25a. [score:3]
Figure S3 miR-153 targets snap-25b. [score:3]
miR-153 Regulation of Motor Neuron Development. [score:3]
Co-injection of morpholinos against both miR-153 and SNAP-25 largely restored normal secondary motor neuron patterning, although the injection of snap-25a,b mRNAs was not as effective at rescuing the defects that resulted from miR-153 overexpression (Fig. S6). [score:3]
0057080.g006 Figure 6 miR-153 regulates primary motor neuron development. [score:3]
0057080.g002 Figure 2 miR-153 targets snap-25a. [score:3]
In zebrafish, miR-153 is expressed in distinct regions of the developing nervous system and brain, including neurosecretory cells of the hypothalamus [29], [30]. [score:3]
We found that knockdown of miR-153 caused a significant increase in GFP expression compared to embryos with wild type levels of endogenous miR-153. [score:3]
These results indicate specific targeting of snap-25 by miR-153. [score:3]
To ensure that the effects of miR-153 on motor neuron patterning were due to expression of miR-153 in these cells, we FACS sorted cells from the trunks of 52 hpf (Tg(mnx1:TagRFP-T) embryos and conducted RT/qPCR. [score:3]
Deletion of both MREs from snap-25a and all three MREs from snap-25b abolished the ability of miR-153 to silence expression (Fig. 2B; Fig. S3B). [score:3]
For movement, exposure to BoNT A rescued the hyperactive phenotypes observed after injection with MOs against miR-153 or overexpression of snap-25a&b mRNAs (Fig. 4C; Movie S1). [score:3]
0057080.g007 Figure 7 miR-153 is expressed in motor neurons. [score:3]
Expression of miR-153 in Motor Neurons. [score:3]
Compared to NIC labeling, miR-153 overexpression resulted in a significant decrease in FM1-43 loading in presynaptic terminals, indicating slowing of the SV cycle (Fig. 8B). [score:2]
0057080.g001 Figure 1 miR-153 regulates embryonic movement. [score:2]
To further dissect the function of miR-153 on motor neuron development, immunofluorescence was performed on whole-mount zebrafish embryos (55 hpf) with antibodies that label primary (Znp-1 or anti-synaptotagmin 2) or secondary (Zn-8 or Alcama) motor neurons [47]. [score:2]
miR-153 Regulates Embryonic Movement. [score:2]
miR-153 regulates the morphology and structure of motor neurons. [score:2]
In contrast, knockdown of miR-153 caused a dramatic and significant 7-fold increase in the frequency of spontaneous movement (Fig. 1). [score:2]
Compared with NICs, overexpression of miR-153 dramatically changed the axonal architecture with significant decreases in branch numbers and length (Fig. 5C, D). [score:2]
These data strongly support the conclusion that miR-153 functions to precisely control SNAP-25 levels to regulate vesicle exocytosis. [score:2]
Figure S4 Dose -dependent rescue of miR-153 knockdown. [score:2]
In sharp contrast, knockdown of miR-153 showed a significant increase in FM1-43 loading, indicating an elevated SV cycling rate (Fig. 8C). [score:2]
miR-153 regulates embryonic movement. [score:2]
To determine the function of miR-153, we injected either synthetic miR-153 or antisense morpholinos against miR-153 into single cell embryos and allowed development to proceed for 1–2 days. [score:2]
A significant decrease in branching was observed in miR-153 injected embryos whereas knockdown of miR-153 caused a dramatic increase in branching. [score:2]
miR-153 Regulates Vesicular Exocytosis to Control Signaling. [score:2]
0057080.g009 Figure 9 miR-153/snap-25 regulates vesicular exocytosis. [score:2]
miR-153 Regulates Embryonic Movement miR-153 has been proposed to be one of a limited number of ancient miRNAs that evolved with the establishment of tissue identity [28]. [score:2]
Knockdown of miR-153 resulted in completely opposite effects with increased motor projection architectural complexity, increased axonal length, and increased branch numbers (Fig. 5B–D). [score:2]
miR-153 Regulates snap-25 to Control Movement. [score:2]
We therefore sought to determine whether snap-25 regulation by miR-153 would alter neuronal morphogenesis. [score:2]
Together, these results reveal a key function for miR-153 in the control of presynaptic vesicle release at the embryonic NMJ, consistent with a role for miR-153 in the regulation of embryonic movement. [score:2]
Under these conditions, excess miR-153 led to a ∼50% decrease in SNAP-25 levels whereas knockdown of endogenous miR-153 increased SNAP-25 levels ∼2-fold. [score:2]
miR-153 regulates synaptic activity at the neuromuscular junction. [score:2]
miR-153/snap-25 regulates vesicular exocytosis. [score:2]
Taken together, the in vivo reporter assays and western blots support the conclusion that snap-25 is a target of miR-153. [score:2]
0057080.g008 Figure 8 miR-153 regulates synaptic activity at the neuromuscular junction. [score:2]
0057080.g005 Figure 5 miR-153 regulates the morphology and structure of motor neurons. [score:2]
miR-153 Regulates Vesicular Exocytosis to Control SignalingSince SNAP-25 has a well-established function in the fusion and release of numerous vesicle types, we next examined the role that miR-153 plays in modulating exocytosis. [score:2]
We first injected miR-153 or morpholinos against miR-153 to observe the effects on the development and morphology of motor neurons in a transgenic zebrafish line in which motor neurons are specifically labeled with RFP (Tg(mnx1:TagRFP-T) [46]. [score:2]
Effects of Knockdown of miR-153 on Movement at 24 hpf Single cell zebrafish embryos were injected with miR-153 [MOs] and filmed for one minute at 24 hpf. [score:2]
Transfections were performed with 300 nM miR-153, biotinylated snap-25 MOs and miR-153 MOs and 1.5 µg of snap-25a,b using Lipofectamine 2000 [84]. [score:1]
Effects of Botulinum Exposure and co-Injection of miR-153 [MO] on Movement at 28 hpf Single cell zebrafish embryos were injected with miR-153 [MOs] and treated with Botulinum toxin A at 27 hpf. [score:1]
Synthetic mRNAs prepared from these reporters were injected into single cell embryos in the presence or absence of exogenous miR-153 or miR-153 morpholinos (MOs). [score:1]
It is possible that the requirement for SNAP-25 may be species specific but we found that altered levels of miR-153 caused similar branching defects in rat PC12 cells as observed in zebrafish motor neurons, strongly arguing against this (data not shown). [score:1]
Interestingly, upon touch stimulation, miR-153 morphants would initially respond with unusually robust, hyperactive movements after which all motion would cease altogether for a period of time (whether touched or not), followed by a resumption of hyperactive movement upon stimulation. [score:1]
The miR-153 sequence is indicated in red and the corresponding snap-25a UTR sequence is shown in green. [score:1]
Owing to the core role of miR-153 in movement control, we first focused on synaptic activity at the neuromuscular junction (NMJ) in zebrafish embryos. [score:1]
miR-153 Regulation of Motor Neuron DevelopmentSNAP-25 is a well-characterized t-SNARE protein, with an established function in vesicular exocytosis [1]– [3]. [score:1]
Despite different levels of conservation, both MREs in snap-25a pair extensively with miR-153 in the seed region. [score:1]
To test for specificity we co -injected embryos with combinations of miR-153, snap-25a,b mRNAs, or morpholinos against both (Fig. 3). [score:1]
Co-injection experiments showed that snap-25a,b mRNA and morpholinos against snap-25 could partially counteract the effects of the corresponding gain and loss of miR-153. [score:1]
Embryos were injected in the presence or absence of exogenous miR-153 or morpholinos against miR-153 (miR-153 [MO]). [score:1]
Single cell embryos were injected with either miR-153 or antisense morpholinos followed by western blots on pooled 1 dpf embryo lysates using antibodies against SNAP-25. [score:1]
The exact pairings between the MREs and miR-153 are shown in Figure 2 and Figure S3. [score:1]
miR-153 has been proposed to be one of a limited number of ancient miRNAs that evolved with the establishment of tissue identity [28]. [score:1]
Perturbation of miR-153 levels caused striking changes in motor neuron structure and branching (Fig. 5A,B). [score:1]
Similarly, co-injection of morpholinos against both snap-25 and miR-153 also restored normal movement (Fig. 1; Movie S1). [score:1]
Strikingly, embryos injected with miR-153 were almost completely motionless, with little or no spontaneous movement, although their hearts were beating normally and minimal movement could be elicited by touch stimulation (Fig. 1). [score:1]
0057080.g004 Figure 4 miR-153 mimics the effects of BoNT A. (A) Single cell embryos were injected as indicated and then at 27 hpf, exposed to Botulinum neurotoxin A (BoNT) for 30 minutes. [score:1]
Because zebrafish motor neuron development is well characterized [40]– [45], we decided to focus on the effects of miR-153 on motor neurons during early zebrafish development. [score:1]
The differences observed due to perturbation of miR-153 levels in the GH4C1 cell line compared to embryonic NMJs are most likely due to differences in the efficiency of miR-153/ miR-153 [MO] delivery between the two experiments, as well as developmental differences. [score:1]
Effects of co-Injection of miR-153 [MO] and snap-25a,b [MO] on Movement at 24 hpf Single cell zebrafish embryos were co -injected with miR-153 [MO] and snap-25a,b [MO] and filmed for one minute at 24 hpf. [score:1]
Since SNAP-25 has a well-established function in the fusion and release of numerous vesicle types, we next examined the role that miR-153 plays in modulating exocytosis. [score:1]
For rescue experiments, co-injection of snap-25a,b mRNA with miR-153 restored near normal movement (Fig. 1; Movie S1). [score:1]
Single cell zebrafish male and female embryos were injected with 200 pg of miR-153, 5 ng each of miR-153 [MO] and miR-153 [loopMO] and/or 100 pg of in vitro-transcribed, capped GFP reporter mRNA with or without the snap-25a or b 3′UTR. [score:1]
Two different morpholinos against miR-153 were utilized. [score:1]
Effects of co-Injection of miR-153 and snap-25a,b on Movement at 24 hpf Single cell zebrafish embryos were co -injected with miR-153 and snap-25a,b mRNA and filmed for one minute at 24 hpf. [score:1]
RNA was isolated from these cell fractions and RT/PCR was performed to determine miR-153 levels relative to U6 snRNA. [score:1]
Precise control of SNAP-25 by miR-153 is necessary not only for presynaptic vesicle release, but also for protein secretion, motor neuron patterning, and outgrowth. [score:1]
The 3′ UTRs from mouse, human and zebrafish snap-25a (A) and snap-25b (B) are shown with the MREs that pair with miR-153 boxed in green. [score:1]
miR-153 mimics the effects of BoNT A.. [score:1]
In contrast, co-injection of miR-153 and snap-25a,b mRNAs or morpholinos against miR-153 and snap-25a,b almost completely restored the normal patterning and branching of motor neurons (Fig. 5B–D). [score:1]
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[+] score: 2
The results of miRror2.0 for the input of mmu miR-98, mmu miR-124, mmu miR-153 and mmu miR-361 are shown. [score:1]
Figure 2A shows the difference in the mapping of the four selected mouse miRNAs (mmu-miR-124, mmu-miR-153, mmu-miR-361 and mmu-miR-98; only four miRNAs were selected for simplicity). [score:1]
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[+] score: 2
Given that brain morphogenesis disruption by ethanol exposure resulted in abnormal neurobehavioral development in zebrafish larvae and juveniles, miRNAs were essential for the establishment of vertebrate neurobehavioral and skeletal systems (e. g., miR-9/9* and miR-153c) [10]. [score:2]
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[+] score: 1
Other miRNAs from this paper: dre-mir-153b, dre-mir-153a
Developing fish were placed in a Sanyo MIR-153 incubator (Amsterdam, The Netherlands) with heating and cooling capabilities for maximum temperature stability. [score:1]
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