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4 publications mentioning dre-mir-153b

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

1
[+] 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: 5
Other miRNAs from this paper: dre-mir-181b-1, dre-mir-181b-2, dre-mir-181a-1, dre-let-7a-1, dre-let-7a-2, dre-let-7a-3, dre-let-7a-4, dre-let-7a-5, dre-let-7a-6, dre-let-7b, dre-let-7c-1, dre-let-7c-2, dre-let-7d-1, dre-let-7d-2, dre-let-7e, dre-let-7f, dre-let-7g-1, dre-let-7g-2, dre-let-7h, dre-let-7i, dre-mir-9-1, dre-mir-9-2, dre-mir-9-4, dre-mir-9-3, dre-mir-9-5, dre-mir-9-6, dre-mir-9-7, dre-mir-16b, dre-mir-16c, dre-mir-29a, dre-mir-101a, dre-mir-144, dre-mir-181c, dre-mir-462, dre-mir-457b, dre-let-7j, dre-mir-181a-2, dre-mir-1388, dre-mir-7147, ipu-let-7a-7, ipu-let-7a-1, ipu-let-7a-3, ipu-let-7a-5, ipu-let-7a-6, ipu-let-7a-4, ipu-let-7a-2, ipu-let-7b-2, ipu-let-7b-1, ipu-let-7c-1, ipu-let-7c-2, ipu-let-7d-2, ipu-let-7d-1, ipu-let-7e-2, ipu-let-7e-1, ipu-let-7f, ipu-let-7g-1, ipu-let-7g-2, ipu-let-7h, ipu-let-7i, ipu-let-7j-1, ipu-let-7j-2, ipu-mir-101a, ipu-mir-1388, ipu-mir-144, ipu-mir-153b, ipu-mir-16b, ipu-mir-181a-1, ipu-mir-181a-2, ipu-mir-181a-3, ipu-mir-181a-4, ipu-mir-181a-5, ipu-mir-181b-2, ipu-mir-181b-1, ipu-mir-181c, ipu-mir-462, ipu-mir-9-4, ipu-mir-9-2, ipu-mir-9-6, ipu-mir-9-1, ipu-mir-9-3, ipu-mir-9-7, ipu-mir-9-5, ipu-mir-7147, ipu-mir-29a, ipu-mir-16c, ipu-mir-203c, ipu-mir-129b, ipu-mir-7553, ipu-mir-7556, ipu-mir-7562, ipu-mir-7568, ipu-mir-7569, ipu-mir-7570, ipu-mir-7571, ipu-mir-7572, ipu-mir-7573, ipu-mir-7574, ipu-mir-7575, ipu-mir-7576, ipu-mir-7577, ipu-mir-457b, dre-mir-181a-4, dre-mir-181a-3, dre-mir-181a-5, dre-mir-181b-3, dre-mir-181d
Therefore, the relatively high number of reads of ipu-miR-144*, ipu-miR-153b* and ipu-miR-16b* indicates that they may play a functional role in regulating gene expression. [score:4]
In channel catfish, most of the miRNA*s were detected at low levels except for ipu-miR-144*, ipu-miR-153b* and ipu-miR-16b* (Table S1). [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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4
[+] score: 1
Other miRNAs from this paper: dre-mir-153a, dre-mir-153c
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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