1. Lectures 10, 11 and Paper 1 Rachel A. Kaplan and Elbert Heng 3.11.14

2. Announcements You have an exam next week, get amped Spring break is after your exam next week, also become accordingly amped You should have read Paper 1 for today’s section It’s my birthday one month from last Sunday, so if you want to get me a present…

3. LECTURE 10: INDIRECT MECHANISMS OF SYNAPTIC TRANSMISSION

4. LECTURE 11: MECHANOSENSATION

5. Mechanosensation Definition: detection of physical movement (of things around, in and on your body) – e.g. touch, stretch, pressure, sound, movement

6. Accomplished by mechanosensory neurons Stretch-gated channels tethered to intra and extracellular matrices – Fast, sensitive, adaptable (so that it can transduce a wide range of inputs), and specialized Lots of receptor subtypes – E.g. Pacinian Corpuscles Respond to vibration because they are fast adapting Neuron is surrounded by epithelial cells that form many layers of gelatinous membranes called lamellae – Pressure on causes neurons to fire – Pressure off also causes neurons to fire

7. C. Elegans is a model It responds to gentle touch on its neurons – ALM, PLM, AVM, PVM – Neuron’s response: inward Na+ current to touch immediately Therefore it is putatively a mechanosensitive neuron Mec-4 is a gene that when mutated eliminates stretch currents (and therefore touch sensitivity) Mec-10 mutation causes channel to be leaky and causes toxic gain of function – Both Mec-4,10 are channel subunit proteins

8. More Neurons/Proteins Involved Degenerin/ENaC Channels – Respond to stretch/mechanical stimulation – slow adapting CEP Neurons – Senses viscosity of surrounding bacteria – Rapidly adapting cation channel – TRP-4: mechanosensory channel Other TRP Channels – Sense temperature, chemical tastants

9. Hearing and Proprioception Vibrations of air are transduced by mechanosensory hair cells – Stereocilia are deflected, links between stereocilia are stretched, allows K+ inward current to depolarize cell Deflecting the other way will hyperpolarize the hair cell Stereocilia adapt by tightening tip links Movement of head in space is transduced by similar hair cells in other organs – Utricle and sacculus – linear acceleration moves gel and crystals, causes opening of hair cells – Semicircular canals – rotational motion causes fluid in canals to move ampulla and embedded hair cells

10. More on hearing… Basilar membrane moves hair cells against tectorial membrane, that moves stereocilia Outer hair cells tension the tectorial membrane - amplify vibrations – OHC depolarized by movement, their depolarization makes them change their size, them changing their size moves the tectorial membrane which causes more depolarization in OHC and IHC

11. PAPER 1: GLUTAMATE-MEDIATED EXTRASYNAPTIC INHIBITION: DIRECT COUPLING OF NMDA RECEPTORS TO CA 2+-ACTIVATED K+ CHANNELS

12. In a nutshell… Putative K+ Outward Current from BK Channels Putative Ca2+ and Na+ Inward Current from NMDARs This is necessary for this

13. Introduction NMDARs are Glutamate Receptors – Causes Ca2+ and Na+ influx – Mg2+ block membrane when hyperpolarized – Ca2+ is a 2 nd messenger and is crucial for plasticity (LTP, LTD) BK Channels are Ca2+ Activated Channels – Causes K+ efflux which hyperpolarizes membrane – Alters neuronal excitability

14. Hypothesis Ca2++ influx from NMDAR activation in turn activates BK channels, which makes it more difficult to depolarize the cell, affecting neuronal excitability. – A new function for NMDARs!

15. Figure 1 A1, A2

16. Figure 1B, 1C (not shown) NMDAR BlockK+ Block

17. Figure 1D

18. Additional Experiment Between Figures… Glu generated largest inhibitory currents when applied to soma but no inhibitory currents when applied to dendrites! – Conclusion: this special phenomenon of NMDAR mediated outward current is localized

19. Fig 2A, 2B

20. Figure 3 Step current (outward) NMDAR mediated current (outward)

21. Figure 4

22. Figure 5

23. Figure 6

24. Figure 7

25. Figure 8A hyperpolarization depolarization

26. Figure 8B