Imaging Neuronal Activity in the Optic Tectum of Late Stage Larval Zebrafish

Bergmann, Katharina; Santoscoy, Paola Meza; Lygdas, Konstantinos; Nikolaeva, Yulia; Macdonald, R.; Cunliffe, Vincent T.; Nikolaev, Anton

doi:10.3390/jdb6010006

Cited by 29 publications

(31 citation statements)

References 47 publications

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“…When the neural activity reporter 256 transgene NBT:GCaMP3 was introduced into the stxbp1a sh438 mutant line, imaging 257 of GCaMP3 fluorescence in 3 d.p.f. larvae(Bergmann et al, 2018) further confirmed 258…”

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confidence: 68%

“…When they were aged 3 d.p.f., transgenic embryos that 721 were motile or immotile were then mounted in 3% agarose in the Zeiss Z.1 Light 722 Sheet microscope and imaged for 5 minutes. Imaging data was processed and 723 analysed with an algorithm to measure GCaMP3 fluorescence levels within larval 724 brain Z-sections (Bergmann et al, 2018). The authors declare that they have no competing interests.…”

Section: Extraction and Quantification Of Cortisol 574mentioning

confidence: 99%

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Regulation of neuron-specific gene transcription by stress hormone signalling requires synaptic activity in zebrafish

Eachus¹,

Subramanya²,

Jackson³

et al. 2019

Preprint

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28The Glucocorticoid Receptor (GR) co-ordinates metabolic and behavioural 29 responses to stressors. We hypothesised that GR influences behaviour by 30 modulating specific epigenetic and transcriptional processes in the brain. Using the 31 zebrafish as a model organism, the brain methylomes of wild-type and gr s357 mutant 32 adults were analysed and GR-sensitive, differentially methylated regions (GR-DMRs) 33 were identified. Two genes with GR-DMRs exhibited distinct methylation and 34 transcriptional sensitivities to GR: the widely expressed direct GR target fkbp5 and 35 neuron-specific aplp1. In larvae, neural activity is required for GR-mediated 36 transcription of aplp1, but not for that of fkbp5. GR regulates metabotropic glutamate 37 receptor gene expression, the activities of which also modulated aplp1 expression, 38 implicating synaptic neurotransmission as an effector of GR function upstream of 39 aplp1. Our results identify two distinct routes of GR-regulated transcription in the 40 brain, including a pathway through which GR couples endocrine signalling to 41 synaptic activity-regulated transcription by modulating metabotropic glutamate 42 receptor expression. 43 44 45 46 Eachus, Subramanya et al. Results 87A loss-of-function mutation in the nr3c1 gene encoding the zebrafish Glucocorticoid 88Receptor (GR), denoted gr s357 , was previously shown to cause abnormally high 89 levels of HPI axis gene expression, elevated blood cortisol, and behavioural 90 abnormalities suggestive of increased behavioural anxiety (Griffiths et al., 2012; Ziv 91 et al., 2013). An initial analysis of locomotor behaviours and measurement of whole 92 body cortisol of wild-type and gr s357 mutant adults and larvae supported and 93 extended these findings (Figure 1), confirming a requirement for GR in regulation of 94 both nervous and endocrine system functions in larval and adult stages. gr s357 95 mutant larvae and adults swam more slowly, larvae were more dispersed, and adults 96 froze more frequently, were less exploratory and less dark aversive than age-97 matched wild-type animals (Figure 1 A-J). Moreover, gr s357 mutant larvae and adults 98 had higher whole body cortisol levels than age-matched wild-type animals (Figure 1 99

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confidence: 68%

Section: Extraction and Quantification Of Cortisol 574mentioning

confidence: 99%

Regulation of neuron-specific gene transcription by stress hormone signalling requires synaptic activity in zebrafish

Eachus¹,

Subramanya²,

Jackson³

et al. 2019

Preprint

View full text Add to dashboard Cite

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“…We were unable to visualise cytosolic GFP in the majority of kugeln; most were not visible using the cytosolic reporter and were only observed using the membrane-tagged reporter (Figure 3C). To characterise the tissue surrounding the kugeln, we examined double transgenics expressing GFP in neurons of the developing brain (Bergmann et al, 2018), and observed exclusion of neural tissue from kugeln (Figure 3D).…”

Section: Resultsmentioning

confidence: 99%

“…The following zebrafish lines were used; Tg(kdrl:HRAS-mCherry) s916 labels EC membrane (Chi et al, 2008) (AB background (Streisinger et al, 1981); Casper background (White et al, 2008)), Tg(fli1a:eGFP) y1 labels EC cytoplasm (Lawson and Weinstein, 2002), Tg(flk1:nls-eGFP) zf109 labels EC nuclei (Blum et al, 2008), Tg(fli1a:Lifeact-mClover) sh467 labels endothelial filamentous actin (manuscript under review and uploaded to editorial manager), and Tg(nbt:GCaMP3) labels neurons (Bergmann et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Endothelial cells form transient Notch-dependent NO-containing cystic structures during zebrafish cerebrovascular development

Kugler

Chhabria

Daetwyler

et al. 2018

Preprint

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Endothelial cell behaviour during blood vessel formation is highly complex and dynamic. Transgenic zebrafish have provided many new insights into these processes, due to their ability to provide detailed in vivo imaging.We here report a previously undescribed endothelial cell behaviour during zebrafish embryonic development. Endothelial cells of the cerebral vessels of 3-5d post fertilisation embryos extruded large membranous spherical structures. These were only found on the cerebral vessels, and did not detach from the parent vessel, instead regressing back into the endothelial cell. These structures did not communicate with the vessel lumen, exhibited periodic oscillations in size and shape, and were enriched with filamentous actin at their neck. Due to their unknown nature and spherical appearance we termed these structures kugeln (German for sphere).Pharmacological inhibition of vascular endothelial growth factor (VEGF) signalling significantly increased kugel number while Notch inhibition significantly reduced both kugel number and diameter.Kugeln contain little cytoplasm, but are highly positive for nitric oxide (NO) reactivity, suggesting they represent a novel NO containing organelle specific to the cerebral vessels.

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“…We uncovered several avoidance strategies, which indicate the presence of flexible neural mechanisms underlying the behavior. Recently developed techniques for embedding of juvenile zebrafish (Bergmann et al, 2018; Matsuda et al, 2017; Vendrell-Llopis & Yaksi, 2016), combined with immersive virtual reality setups, make juvenile zebrafish a promising model organism for future studies of brain activity in a navigating animal.…”

Section: Discussionmentioning

confidence: 99%

Zebrafish use visual cues and geometric relationships to form a spatial memory

Yashina

Tejero-Cantero

Herz

et al. 2019

Preprint

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15Animals use salient cues to navigate in their environment, but their specific cognitive 16 strategies are largely unknown. We developed a conditioned place avoidance paradigm 17 to discover whether and how zebrafish form spatial memories in a Y-shaped maze. 18 Juvenile zebrafish, older than three weeks, learned to avoid the arm of the maze that 19 was cued with a mild electric shock. We found that the fish required distinct visual 20 patterns to develop a conditioned response. Interestingly, individual fish solve this task 21 in different ways: by staying in the safe center of the maze, by preference for one, or 22 both, of the safe patterns, or by mixed strategies. In experiments in which the learned 23 patterns were swapped, rotated or replaced, the animals could transfer the association 24 of safety to a different arm or to a different pattern using either visual cues or location 25 as the conditioned stimulus. These findings show that juvenile zebrafish exhibit several 26 complementary spatial learning modes and pave the way for neurobiological studies of 27 navigational mechanisms in this model species. 28 29 104 105A biologically realistic agent model replicates learned avoidance behavior 106 Larval and juvenile zebrafish swim in characteristic swim bouts (Supplementary Video 1). 107Electric stimulation caused swim bouts of increased amplitude compared to the amplitude of 108 spontaneous swim bouts ( Figure 1E). We hypothesized that increased swimming speeds 109 under electric stimulation in the conditioned arm could lead to changes in the OC and EF 110 measures independent of the learning abilities of the fish. We developed a null-model of the 111 6 112 Figure 1. The conditioned place avoidance (CPA) paradigm.113 (A) Experimental setup. (B) Measures of fish performance in the paradigm, top: arm occupancy (OC); bottom: 114 arm entry frequency (EF); middle: schematic for the calculation of the preference scores for OC/EF. Positive 115 values of the scores correspond to arm preference, negative values correspond to arm avoidance. Red color 116 represents the shocked arm, black and gray represent the safe arms. (C) Left: moving averages of the OC/EF 117 scores in a 2-hour control experiment (mean ± s.e.m.). Right: box plots for OC/EF scores in the first and the last 118 5 minutes of the control experiments (Mann-Whitney test, n = 24 fish). Box plots show median and quartiles; 119 whiskers show 1.5x interquartile range; dots show values for individual fish. (D) There is no significant 120 preference for any of the visual patterns (ANOVA, n = 24 fish). (E) Distributions of amplitudes for spontaneous 121 (gray) and shock-triggered swim bouts (red). (F) Pseudo-random 1D walk model used to evaluate CPA 122 measures. Bout amplitude S for simulated movement was drawn from a Gamma distribution, amplitude in the 123 conditioned arm was multiplied by a speed ratio α. (G) Top: model without learning. Bottom: model with 124 learning. Learning rule was implemented by decreasing the probability of entry into t...

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Imaging Neuronal Activity in the Optic Tectum of Late Stage Larval Zebrafish

Cited by 29 publications

References 47 publications

Regulation of neuron-specific gene transcription by stress hormone signalling requires synaptic activity in zebrafish

Regulation of neuron-specific gene transcription by stress hormone signalling requires synaptic activity in zebrafish

Endothelial cells form transient Notch-dependent NO-containing cystic structures during zebrafish cerebrovascular development

Zebrafish use visual cues and geometric relationships to form a spatial memory

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