1. Non-striated muscle II

3. Hospital Volunteer Wanna-bes Pullman Regional Hospital is now accepting volunteer applications for the spring semester. Applications are due no later than Wednesday, January 18, 2012. Students may find more information about volunteering and the application requirements at https://www.pullmanregional.org/wsu-student- volunteers. https://www.pullmanregional.org/wsu-student- volunteers With best regards, Kim Cook

4. Excitation-contraction coupling: Two sources of Ca ++ for smooth muscle contraction Extracellular: through L Ca ++ channels Intracellular: from cell-surface associated SR (calciosomes)

5. Two routes for excitation-contraction coupling Route 1: Electromechanical: depolarization opens L Ca ++ channels in sarcolemma; L channels open nearby calciosome release channels (ryanodine receptors) by Ca ++ - induced Ca ++ release. Each L channel thus ignites a calcium spark.

6. Route 2: Pharmacomechanical: transmitter or hormone sends 2nd message to calciosomes (and possibly also to L channels) to induce Ca ++ release/entry.

7. Contraction or relaxation? It all depends on the 2nd message(s) the muscarinic receptor is coupled to Ach stimulates (constricts) airway GI tract pupillary constrictors Ach inhibits (dilates) specialized (genital) vasculature myocardium

8. Ach causes action potentials and contraction in toad gastric smooth muscle. The voltage is measured by the microelectrode. The letters above the trace indicate the timing of the video frames.

9. A DAG-IP 3 2nd Message: contraction of smooth muscle by acetylcholine Step 1. Acetylcholine released by the parasympathetic branch of the autonomic nervous system, binds to the muscarinic receptor for acetylcholine of intestinal smooth muscle. Step 2: The combination of neurotransmitter and receptor activates G protein

10. Step 3: G protein activates membrane- bound Phospholipase C Step 4: Phospholipase C converts some phosphatidylinositol bisphosphate (a membrane phospholipid) into inositol trisphosphate (IP 3 ) and diacylglycerol. (DAG)

11. Phospholipase C splits off the “tails” of the phospholipid as DAG, and adds a phosphate to the “head” to form IP 3

12. Step 5: Now the story splits into two directions. Direction 1: Still in the plasma membrame - DAG remains in the membrane. It activates Protein Kinase C, which has to translocate into the membrane to become active. Once PKC is active, it can phosphorylate many proteins, both membrane bound and free.

13. Direction 2: In the cytoplasm - IP 3 diffuses through the cytoplasm. It binds to a receptor on the endoplasmic reticulum, a storage site for Ca ++, causing Ca ++ release.

14. Final Effects : Through PKC, DAG activates membrane channels that result in depolarization, with a change in the basic electrical rhythm. Through IP 3, Ca ++ is released from internal stores and the force of contraction is increased.

15. How the muscarinic receptor excites GI smooth muscle 1. Muscarinic receptor coupled to IP 3 -DAG 2nd message. 2. DAG activates protein kinase C 3. PKC inhibits rest K + channels, causing depolarization. 4. Depolarization activates L channels 5. L channels activate Ca ++ release from calciosomes. The electromechanical route:

16. The pharmacomechanical route 1. Muscarinic receptor coupled to IP 3 - DAG 2nd message. 2. IP 3 activates release of Ca ++ from calciosomes

17. A classic experiment with intestinal smooth muscle Remove length of intestine from animal Tie one end to rigid support Attach other end to force transducer Submerge muscle in heated, oxygenated saline.

18. The anatomy of the intestinal muscularis Myenteric plexus (yellow) Submucosal plexus (orange) Longitudinal muscle outer layer) – dominated by electromechanical route Circular muscle (inner layer) – dominated by pharmacomechanical route Interstitial cells of Cajal (green)

19. The preparation is spontaneously active; acetylcholine both shortens the muscle and increases the magnitude of the oscillations. Isotonic length 1 min Upward displacements of the trace correspond to decreases in length

20. Isotonic length Ca ++ -free saline can be used to dissect the electromechanical and pharmacomechanical mechanisms Spontaneous contractions are eliminated in Ca ++ -free saline, which abolishes the electromechanical route; addition of acetylcholine can still elicit episodes of contraction by the pharmacomechanical route, but these diminish in magnitude with each subsequent addition (why?).

21. Epinephrine abolishes spontaneous activity and causes the muscle to go to a longer length Isotonic length

22. How the beta 2 receptor turns off contraction in GI smooth muscle 1.  2 receptor is coupled to cAMP 2nd message. 2. cAMP activates Protein kinase A 3. PKA phosphorylates MLCK 4. Phosphorylated MLCK cannot be activated by calmodulin-Ca ++

23. MLCK-P (inactive) PKA Beta adrenergic receptor cAMP Norepinephrine

24. There are many endogenous transmitter substances in the gut Excitation –Ach –Serotonin –Peptides (motilin) Inhibition –catecholamines –Vasoactive Intestinal Peptide/nitric oxide –Dopamine –Endogenous opiates

25. Smooth muscle in the bedroom

26. NOS = nitric oxide synthase G-cyclase = guanylyl cyclase G-kinase =cGMP dependent protein kinase L-nmma and L- name are inhibitors of NOS O 2 and HbO 2 get rid of NO by oxidizing it

27. Let’s get to the point – some anatomy In the flaccid penis (and clitoris), cavernous arteries are constricted and the small amount of volume flow into the sinuses easily drains into the veins. Erection is the result of dilation of the cavernous arteries and trabecular smooth muscle. As the sinuses fill with blood under arterial pressure, they compress the vessels in the venular plexus, making outflow more difficult.

28. How para- sympathetic input causes erection

29. How sympathetic input causes you to lose it, or not even get it Smooth muscle contraction = vascular sinus deflation

30. The understanding of the nature and origin of EDRF as NO led rapidly to the development of drugs that act on this system. For example, Viagra and its congeners act by inhibiting a form of cGMP phosphodiesterase that is specific to genital vasculature. The drugs are not perfectly specific, so side effects include hypotension and a mild impairment of color vision (remember that phototransduction also involves a cGMP 2 nd messenger, so you can see why effects on vision might occur). Patients who take nitroglycerine for their angina should not also take Viagra – why not?