Lecture: 4 – Synapses and Synaptic Transmission Date: August 3, 2015
Lecturer: Dr. Arlene R. Ng, M.D., Ph.D. Trans Team: Diaz, Dino, Endaya
Topic Outline o With direct open fluid channels conducting electricity
from one cell to the next I. Synapses and Synaptic Transmission A. Definition of a Synapse o Pre- and postsynaptic membranes come close B. Synaptic Transmission together forming gap junctions II. Types of Synapses o Form low-resistance bridges through which ions A. Electrical Synapse pass with relative ease from the interior of one cell B. Chemical Synapse to the interior of the next III. Principles on Chemical Transmission Gap junctions and connexons A. Neurotransmitters o Direct channels (connexon) are low-resistance B. Presynaptic Cell Processes pathways C. Synaptic Cleft Processes o Connexins (proteins) form connexons (which are D. Postsynaptic Cell Processes porous at the junctions) IV. Overall process o Mediating agent: ionic current V. Synaptic Delay VI. Synaptic integration o Cells are electrically coupled VII. Organization of Neural Pools o Responsible for ephaptic / electrotonic / electrical VIII. Importance of Synapses transmission o Almost instantaneous production of an action potential in the post-synaptic neuron: responsible for I. Synapses and Synaptic Transmission fast escape reflexes o For highly synchronized activity in the brain A. Definition of a Synapse o Particularly common during early embryonic stages Junction where the axon or some other portion of one cell (coordinated growth and maturation of neighboring terminates on the dendrites, soma or axon of another cells) neuron or a muscle or gland cell o Gap junctions are also found in Specialized site of contact - Glia B. Synaptic Transmission - Epithelial cells Process of information transfer at synapses: may block - Smooth muscle cells transmission, changed from a single impulse into repetitive - Cardiac muscle cells impulses or may be integrated with impulses from other - Liver cells neurons - Some glandular cells Complex process – excitatory or inhibitory (or sum of both) - Schwann cells (forming myelin sheath) o Happens whether or not an action potential is generated Wrapped around axons; almost devoid of Permits grading and adjustment of neural activity necessary cytoplasm for normal functions Mediate both intra- and intercellular communication; also serve as nourishment for II. Types of Synapses the axons beneath Passage of ions and metabolites across layers Table 1. Distinguishing properties of Electrical and Chemical Synapses. o Charcot-Marie-Tooth Disease (CMTX) [X Type of Synapse Electrical Chemical Chromosome-linked form] Synapse Synapse o Demyelination due to mutation in connexin32 gene Distance between pre- 3.5nm 20-40nm o Prevents formation of functional gap junction and postsynaptic cell channels essential for normal flow of metabolites in membranes Schwann cells Cytoplasmic continuity Yes No B. Chemical Synapse between pre- and The most prevalent synaptic transmission in the postsynaptic cells mature human nervous system Ultrastructural Gap- Presynaptic The presynaptic cell contains an abundance of components junction vesicles and mitochondria, microtubules, and synaptic vesicles channels active zones; The postsynaptic cell has corresponding receptors postsynaptic Secretes neurotransmitters that in turn acts on receptors receptor proteins in the membrane of the next neuron Agent of Transmission Ion current Chemical (may excite, inhibit or modify the second neuron) transmitter One-way conduction: from the presynaptic neuron to Synaptic Delay Virtually Significant: at the post-synaptic neuron. Hence, signals are directed absent least 0.3ms, at very specific goals usually 1-5ms or longer Direction of transmission Usually Unidirectional Bidirectional
A. Electrical Synapse
Figure 1. Electrical Synapse and Gap Junctions.
Page 1 of 5 4 – SYNAPSES AND SYNAPTIC TRANSMISSION
PHYSIOLOGY BLOCK 1
With synaptic cleft
With synaptic delay Abundant Mediating agent: chemical messenger Responsible for chemical transmission
Figures 2 & 3. Anatomy of a Chemical Synapse.
Anatomy of a Chemical Synapse
o Synaptic cleft – 20-40nm Filled with matrix of fibrous extracellular protein (adhesion) o Synaptic vesicles – 50nm o Secretory granules – 100nm Contain secretory proteins Dense core vesicles (appear dark on EM) o Membrane differentiations Dense accumulations of proteins adjacent to Figure 6. Comparison between Delay in Electrical and Chemical Synapses. and within membranes on either side of the synaptic cleft Active zones III. Principles in Chemical Transmission Postsynaptic density A. Neurotransmitters Categories of CNS Synaptic membrane Characteristics: differentiations o Must be synthesized and stored in the presynaptic o Gray’s Type I – Asymmetrical membrane terminal or neuron differentiations = excitatory (enhanced signal to o Must be released upon stimulation ensure exposure) o When experimentally applied, must produce a o Gray’s Type II – Symmetrical membrane response in the postsynaptic cell that mimics the differentiations = inhibitory response produced by release of neurotransmitter from the presynaptic neuron o Existence of removal mechanisms (for recycling) – since not completely used up Disease: Myasthenia gravis - postsynaptic membrane of neuromuscular junction has too few acetylcholine receptors (ACh are degraded by cholinesterases) o Treatment by introduction of anticholinesterase drug to slow acetylcholine breakdown and allow ACh to accumulate Around 50 discovered so far, more not yet elucidated Types: 1. Small-molecule, rapidly acting transmitters a) Class I i. Acetylcholine b) Class II: The Amines i. Norepinephrine ii. Epinephrine iii. Dopamine iv. Serotonin v. Histamine Figure 4. Categories of CNS Synaptic Membrane Differentiation. c) Class III: Amino Acids i. Gamma-aminobutyric acid (GABA) Common sites of synapses: axosomatic, axodendritic ii. Glycine (spine and shaft) and axoaxonic iii. Glutamate iv. Aspartate d) Class IV i. Nitric Oxide (NO) 2. Neuropeptides – slowly acting transmitters or growth factors - Has prolonged action and enacts long-term changes (unlike small molecules) - Synthesized as integral parts of large protein molecules - 1000 times more potent a. Hypothalamic-releasing hormones i. Thyrotropin-releasing hormone ii. Luteinizing hormone-releasing hormone iii. Somatostatin (growth hormone inhibitory factor)
Figure 5. Common Synapse Sites. b. Pituitary peptides
i. Adrenocorticotropic hormone (ACTH)
Page 2 of 5 4 – SYNAPSES AND SYNAPTIC TRANSMISSION
PHYSIOLOGY BLOCK 1
ii. β-endorphin o SNARE (SNAP Receptor; Soluble NSF Attachment
iii. α-melanocyte-stimulation hormone Protein Receptor) Proteins: iv. Prolactin a. V-SNARE (vesicular SNARE) v. Luteinizing hormone Synaptobrevin (VAMP-vesicle vi. Thyrotropin associated membrane protein vii. Growth hormone Synaptotagmin viii. Vasopressin b. T-SNARE (target membrane SNARE) ix. Oxytocin Syntaxin and SNAP-25 c. Peptides that act on gut and brain o Synaptotagmin i. Leucine enkephalin ii. Methionine enkephalin iii. Substance P iv. Gastrin v. Cholecystokinin vi. Vasoactive intestinal polypeptide (VIP) vii. Nerve growth factor viii. Brain-derived neurotropic factor ix. Neurotensin x. Insulin xi. Glucagon d. From other tissue i. Angiotensin II ii. Bradykinin iii. Carnosine iv. Sleep peptides v. Calcitonin B. Presynaptic cell: Processes Figure 8. Mechanism of Vesicular Proteins Synthesis of transmitters o Manufacturing of chemical messengers Mobilization, trafficking, docking of vesicles: Role of o Synthesis located very near the terminals calcium and Rab3A protein o Storage of transmitters in vesicles: Packaging of o Entry of Ca2+ into the nerve terminal leads to the neurotransmitters into vesicles or granules opening of the fusion pore complex and o Release of transmitters, and recycling of vesicles neurotransmitter release. Calcium entry also frees o The chemiosmotic theory: requirement of energy for vesicles from the storage compartment through transport of vesicles (since the act is against the phosphorylation of synapsins thus increasing the concentration gradient availability of vesicles for docking at the presynaptic Storage of transmitters in vesicles plasma membrane. o Packaging of neurotransmitters into vesicles or o The Rab3A cycle targets vesicles to their release granules sites. Rab3A complexed to GTP binds to synaptic o Vesicle Types: vesicles. During the targeting of synaptic vesicles to a. Small, Clear, Spheroid - Acetylcholine, the active zone, Rab3A hydrolysis may serve to GABA, Glycine, Glutamate make a reversible reaction irreversible, preventing b. Small with dense core - Catecholamines, vesicles from leaving the active zone once they serotonin arrive. During fusion and exocytosis, Rab3A-GDP c. Large with dense core - peptides dissociates from the vesicle. There is then an exchange of GTP for GDP. This is followed by the Release of transmitters into the synaptic cleft association of Rab3A-GTP with a new synaptic 1. Development of Action Potential – depolarizes the vesicle thus completing the cycle. presynaptic membrane 2. Opening of voltage-gated calcium channels Molecular Basis for fusion and exocytosis: The SNARE 3. Entry of calcium hypothesis 4. Binding of calcium with release sites o Vesicle and target membrane compartments have 5. Binding of vesicles with release sites (NB: each distinct receptors – the v-SNARES and t-SNARES vesicle contains ~2000-10000 molecules) that mediate docking and fusion. Following fusion, 6. Exocytosis two cytoplasmic proteins, NSF (Neurexin/N- 7. Recycling of transmitter vesicles Ethylmaleimide-sensitive Factor) and SNAP (Soluble NSF Attachment Protein), bind to the SNARE complex and disassemble it
C. Synaptic Cleft: Processes
Removal of transmitters via: o Diffusion o Enzymatic destruction o Re-uptake
D. Post-synaptic Cell: Processes
Receptor proteins in the postsynaptic terminal Figure 7. Steps in the Release of Neurotransmitters. o Components Binding component Vesicular proteins Ionophore component o Synapsins Ion channel Synapsin I - Fast event that closes within milliseconds Between synaptic vesicle and cytoskeletal - Either cation channels (usually Na+) or elements anion channels (usually Cl-) o Rab3A, Rab3C - Cation channels: lined up with negatively- Vesicle trafficking within the cell and vesicle charged residues to attract cations and targeting within nerve terminal repel anions (opened by excitatory neurotransmitters) - Anion channels: small for hydrated cations, but big enough to let anions pass (opened by inhibitory neurotransmitters)
Page 3 of 5 4 – SYNAPSES AND SYNAPTIC TRANSMISSION
PHYSIOLOGY BLOCK 1
2nd messenger activators
- With repeated activation, the transmitter, acting through PKA, will phosphorylate, one or more transcriptional regulatory proteins that activate gene expression. Gene activation results in a protein that produces more enduring closure of the channel and changes in neuronal excitability lasting days or weeks. Processes in the postsynaptic membrane o Excitation Opening of Na+ channels Depressed conduction through K+ or Cl- channels, or both Various changes in internal metabolism of the postsynaptic neuron Figure 10. Temporal and Spatial Summation o Inhibition Opening of Cl- channels If the membrane potential rises high enough, an action Increased conductance of K+ out of the potential can be generated through: neuron 1. Spatial summation Activating receptor enzymes inhibiting 2. Temporal summation cellular metabolic activity Two types of Receptors: o Ionotropic Receptors: Directly gated o Metabotropic Receptors: Indirectly gated
Figure 11. Direct Inhibition
IV. Overall Process
Figure 9. Two Types of Gating. Action potential reaches the synaptic knob opening the Ca2+ channels there Electrical events Ca2+ causes the release of neurotransmitters across the synaptic cleft. Table 2. Distinguishing properties of EPSP and IPSP. EPSP IPSP Neurotransmitters bind to receptors in the postsynaptic Excitatory Postsynaptic Inhibitory Postsynaptic Potential membrane causing certain ion channels to open. Potential EPSP Increase excitability Decrease excitability o Na+ and K- channels open. Depolarization Hyperpolarizing response o Na+ rushes in faster than K- rushes out causing the Local response Ionic Basis: increase chloride or inside of the post synaptic membrane to become potassium conductance more positive. Graded Direct inhibition o This depolarization is the Excitatory postsynaptic Non-propagated potential o The EPSP reaches the threshold potential, an Resting Membrane potential action potential is initiated o Valued at -65mV, which is less negative than the IPSP -95mV at the peripheral nerves and in muscle fibers o K+ and or Cl- channels open. o Reason: the presence of ions allow electrical o Outward rush of K+ and/or inward rush of Cl- gradient caused the postsynaptic membrane to become o Consequence: allows variable excitation or less positive (more negative) inhibition o This hyperpolarization is the inhibitory The Nernst Potential: potential (EMF) that exactly postsynaptic potential opposes the movement of a specific ion o The postsynaptic membrane is now less likely to o EMF = ±61 x log ([in]/[out]) (where EMF value is reach the threshold potential, initiation of an action positive for anions and negative for cations) potential is thus inhibited o For Na+: +65mV. but actual EMF is -65mV, hence the V. Synaptic Delay need for a Na-pump to expel Na+ o For K+: -86mV. more negative, so K+ tends to diffuse 0.5ms out of the neurons Interval before a response is obtained in the o For Cl-: -70mV. small difference - very slight influx into postsynaptic neuron after an impulse reaches the the neurons presynaptic terminals Time it takes for synaptic mediator to be released and to act on the membrane of the postsynaptic cell Slowed conduction in polysynaptic pathways
Page 4 of 5 4 – SYNAPSES AND SYNAPTIC TRANSMISSION
PHYSIOLOGY BLOCK 1
VIII. Importance of Synapses
VI. Synaptic Integration 1. Summation 1. Processing of information (learning) 2. Facilitation 2. Storage of information (memory) 3. Inhibition 3. Filtration of information Direct 4. Action of drugs (neuropharmacology) Indirect 5. Causes of mental health disorders o Renshaw cell – inhibits motor neurons adjacent to a motor neuron that is stimulated IX. References (lateral inhibition) o Presynaptic Purbes, Neurosciences (3rd ed.). retrieved from 4. Factors affecting synaptic transmission: http://www.zoology.ubc.ca/.../synapses%20- a) pH %20presynaptic.htm. date of retrieval: 4 August 2015 i. Alkalosis: increases excitability ii. Acidosis: decreases excitability Guyton, A.C. and Hall, J.E. Textbook of Medical Physiology. b.) Hypoxia: decreases excitability (11th ed.) USA: Elsevier Inc. 2006. Chapter 46 c.) Drugs - Caffeine, theophylline, strychnine - Increases excitability X. Quiz
Questions:
1.) True or False: There is no cytoplasmic continuity in the
electrical synapse
2.) True or False: Gray’s type II category of synaptic
membrane differentiations is symmetrical
3.) True or False: Gap Junctions are bridges of high-resistance
through which ions pass with difficulty.
4.) True or False: Connexons are components of the synaptic
cleft. Figure 12. Presynaptic Inhibition and Facilitation. 5.) True or False: Chemical synapse is more abundant as VII. Organization of Neural Pools compared to electrical synapse
1. Convergence 6.) What is CMTX?
2. Divergence 3. Serial processing 7.) What is the agent of transmission in an electrical synapse? 4. Reverberation 5. Parallel – similar to circuits
Answers:
1.) False
2.) True
3.) False
4.) False
5.) True
6.) Charcot- Marie Tooth Disease
7.) Ion current
Figure 13. Organization of Neural Pools
Page 5 of 5 4 – SYNAPSES AND SYNAPTIC TRANSMISSION