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Rafiei, N.

Publications and source records attributed to Rafiei, N..

4 recordsLinked to original sources

Enkephalin constrains fear learning via volume transmission to the lateral amygdala

Fear learning involves the formation of associations between cues and aversive outcomes, a process that must be tightly regulated to prevent excessive or generalised fear. Dopamine release in the lateral amygdala (LA) drives fear acquisition, whereas endogenous opioids constrain it. However, whether opioids are dynamically released within the amygdala circuits during learning, and how they exert this control remain unclear. Here we show that met-enkephalin is locally released within the amygdala during auditory fear conditioning, with signals shifting from the aversive outcome to its predictive cue as learning progresses. The amygdalo-striatal transition zone (ASt), is the principal source of this enkephalin, released from medium spiny neurons receiving strong auditory thalamic input. This enkephalin spreads from the ASt to the LA via volume transmission. Selective knockdown in the ASt abolished opioid signals and enhanced fear learning, demonstrating that this diffuse signal constrains fear memory formation. We further show that enkephalin suppresses dopamine release in both the ASt and LA via {micro}-opioid receptors, identifying the ASt as a neuromodulator hub coordinating opioid and dopaminergic signalling across amygdala fear circuits. Although demonstrated here for auditory fear learning, the ASt receives multimodal sensory input, suggesting a broader mechanism through which sensory experience recruits enkephalin release to gate associative learning

neuroscience↗

Design of a Versatile Microfluidic Device for Imaging Precision-Cut-Tissue Slices

Precision-cut-tissues (PCT), which preserve many aspects of a tissues microenvironment, are typically imaged using conventional sample dishes and chambers. These can require large amounts of reagent and, when used for flow-through experiments, the shear forces applied on the tissues are often ill-defined. Their physical design also makes it difficult to image large volumes and repetitively image smaller regions of interest in the living slice. We report here on the design of a versatile microfluidic device capable of holding mouse or human pancreas PCTs for 3D fluorescence imaging using confocal and selective plane illumination microscopy (SPIM). Our design positions PCTs within a 5 x 5 mm x 140 {micro}m deep chamber fitted with 150 {micro}m tall channels to facilitate media exchange. Shear stress in the device is localized to small regions on the surface of the tissue and can be easily controlled. This design allows for media exchange at flow rates ~10-fold lower than those required for conventional chambers. Finally, this design allows for imaging the same immunofluorescently labelled PCT with high resolution on a confocal and with large field of view on a SPIM, without adversely affecting image quality.

bioengineering↗

Growth Phase Dependent Chromosome Condensation and H-NS Protein Redistribution in E. coli Under Osmotic Stress

The heat-stable nucleoid-structuring (H-NS) protein is a global transcriptional regulator implicated in coordinating the expression of over 200 genes in E. coli bacteria. We have applied super-resolved microscopy to quantify the intracellular, spatial reorganization of H-NS in response to osmotic stress. We find that H-NS shows a growth phase dependent response to hyperosmotic shock. In stationary phase, H-NS detaches from a tightly compacted bacterial chromosome and is excluded from the nucleoid volume over an extended period of time. This behaviour is absent during rapid growth but may be induced by exposing the osmotically stressed culture to the DNA gyrase inhibitor, coumermycin. This compaction/H-NS exclusion phenomenon occurs in the presence of either potassium or sodium ions and is independent of the stress responsive sigma factor RpoS, or the H-NS paralog StpA.

microbiology↗

Resolving Biology Beyond the Diffraction Limit with Single-Molecule Localization Microscopy

Optical imaging provides a window into the microscopic world, but the level of observable detail is ultimately limited by the wavelength of light being employed. By harnessing the physics of photoswitchable dyes and fluorescent proteins, single-molecule localization microscopy (SMLM) provides a window into the nano-world of biology. This mini-review article provides a short overview of SMLM and discusses some of its prospects for the future.

biophysics↗