Tutorial Questions Flashcards
What is the excitation of GFP
Stimulated by blue light. Excitation wavelength of 395nm
Emission of GFP
Emits green light at 509nm wavelength. Microscope imaging tells us about the morphology and function of neurons
Describe channelrhodopsin
It’s a light gated non selective ion channel, which opens upon stimulation by blue light
What is the function of channelrhodopsin
Opening by blue light stimulation causes Na+ influx into cell = depolarisation. If threshold is met, an action potential will be generated
Describe Halorhodopsin
A chloride specific light-gated ion channel, which opens upon yellow light stimulation
What is the function of halorhodopsin
Upon opening by yellow light stimulation, Cl- influx into cell = hyperpolarisation. This moves membrane potential away from threshold, preventing the firing of an action potential
Describe the function of GCaMPs
GCaMPs are GTP-based calcium indicators - allows imaging and visualisation of neuronal responses to different stimuli
Describe the mechanism of GCaMPs
- 2 calcium binding proteins fuse to GFP
- Activity of neurons causes an increase in Ca+ inside the post-synaptic membrane
- In presence of calcium, the 2 binding proteins interact
- This interaction changes confirmation of the GFP
- GFP fluoresces much brighter
What are the benefits of using a confocal microscope
- High spacial resolution (pinhole in between lenses means only light from focal plane Is accepted)
- Can image live samples in situ
- Can produce 3D images
What are the main electrophysiological techniques
- Patch clamp
- Sharp electrode
- RNA tomography
- fMRI
What is the patch clamp technique used for
to study ionic voltage currents of single ion channels in individual isolated living cells, tissue sections, or patches of cell membrane. Opening of sodium channels causes drop in mV- brief downwards deflections in current
Describe how the patch clamp technique is done
- Touch cell with glass pipette filled with electrolyte
- Apply negative pressure to suction membrane
- This forms a tight junction with membrane for ion flow
- Record small electrical current of single channel via electronic amplifier
Patch clamp va sharp electrode
- Only patch clamp can record single channels
- The larger tip of the glass pipette in patch clamp allows for lower resistance = better electrical access to inside of cell
- Patch clamp experiences run-down or dialysis, where sharp electrode can record for longer periods
Describe sharp electrode recordings
uses a fine-tipped glass micropipette inserted into the neuron, allowing direct recording of electrical events generated by the neuron (membrane potential, resistance, time constant, synaptic potentials and action potentials. Records whole channels
Describe RNA tomography
This technique is used to understand neuronal diseases such as stroke. Thinly sliced tissue is profiled in all directions and then mathematically image reconstructed to determine genome wide 3D expression patterns
Describe the structure of the retina
3 layers of neurons and 2 layers of synapses
Layer 1: photoreceptors
Layer 2: outer plexiform layer
layer 3: bipolar cells
Layer 4: inner plexiform layer
Layer 5: ganglion cells
What are the feedforward neurons
- Photoreceptors (excitatory Glu)
- Bipolar cells (excitatory Glu)
- Ganglion cells (inhibitory GABA)
What are the feedback neurons
- Horizontal cells
- Amacrine cells (inhibit ganglion cells)
What are the function of Rods
• active in dim light
• black and white
• low resolution
What are the functions of cones
• active in bright light
• colour
• high resolution - in fovea
What area of the brain processes visual information
The lateral geniculate nucleus - located in the thalamus, it’s responsible for initial processing directly from ganglion cells via optic nerve
What are the 2 visual pathways
- Ventral Stream - identifies objects
- Dorsal Stream - spacial location & speed
Receive information from primary visual cortex
What happens when light intensity increases
• less glutamate release from photoreceptors - cones
• depolarisation of ON bipolar cells in the inner plexiform layer
• hyperpolarisation of OFF bipolar cells
What is the receptive field
An area of the retina which when illuminated activates visual neurons. Indirect activation by horizontal cells
What is centre surround organisation of the receptive fields
allows ganglion cells to transmit information about whether photoreceptor cells are exposed to light and about the differences in firing rates of cells in the center and surround. This is achieved by responding to differences in illumination. Illumination of centre & surround = responses in different polarities
Recall the different ganglion cells
- Parrocellular (80%)
• shape and colour
•Small receptive field - Magnocellular (10%)
• motion detection
•large receptive field
What features of a sound need encoding
- Frequency (pitch)
- Intensity (loudness)
- Onset
- Duration
Need to distinguish different sounds for communication, memory, survival
What are the important steps in transforming sound into a neuronal signal
SOUND WAVES enter the ear canal and cause the eardrum to vibrate. VIBRATIONS pass through 3 connected bones in the middle ear. This motion SETS FLUID MOVING in the inner ear. Moving fluid bends thousands of delicate hair-like cells which convert the vibrations into NERVE IMPULSES.
What are the different chambers of the cochlea
- Scala vestibular
- Scala tympani
- Scala media
What chambers are filled with perilymph
Scala vestibular and Scala tympani
Contains low K+, normal Ca2+ and high Na+
What chambers are filled with endolymph
Scala media
Filled with high K+, low Ca2+ and low Na+
What is the function of the chambers of the cochlea
The ST and SV sense pressure changes caused my sound waves. The SM contains the organ of Corti and basilar membrane where the vibrations move the stereocillia to transduce sound into a a neuronal impulse
What is cochlear tonotopy
In the auditory system their is a gradient of high and low frequency. The apex of the cochlea receives low frequency sounds and the base high frequency sounds