What is the target size range for fragmenting gDNA?
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~100 – 300 bp.
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What is the target size range for fragmenting gDNA?
~100 – 300 bp.
What is another example of a NGS technology?
Ion Torrent sequencing.
What is the primary energy source for exercise lasting 6 to 10 seconds?
ATP from creatine phosphate and ADP (direct phosphorylation).
What gives skeletal muscle its striated appearance?
The arrangement of thick and thin filaments, creating light and dark bands.
What is the purpose of partitioning library fragments in NGS?
To create clusters of identical copies on a solid surface.
What is the function of the collateral ganglion in the Sympathetic Nervous System?
It serves as a relay point for sympathetic fibers to abdominal and pelvic organs.
What is the connective tissue layer surrounding a muscle fiber?
Endomysium.
What does the medulla oblongata most directly control?
Activity of the Autonomic Nervous System (ANS).
What is the principle of Next-Generation Sequencing (NGS)?
Large numbers of DNA samples are sequenced side-by-side on the same apparatus.
What is an example of a NGS technology?
Illumina sequencing.
Can we decide when to digest food?
No, digestion is an involuntary process regulated by the autonomic nervous system.
What energy source is used first during short-duration exercise?
ATP stored in muscles.
What is the sarcoplasmic reticulum (SR) in muscle fibers?
It is the endoplasmic reticulum (ER) that wraps around myofibrils.
How is skeletal muscle attached to the bone?
Via tendon.
What role does Ca2+ play in muscle contraction?
It is essential for the contraction process.
What triggers the exocytosis of synaptic vesicles?
The influx of Ca2+.
How many bases does each cluster decode in NGS?
Between 100 and 300 bases.
What is involved in NGS Library construction?
Fragmenting genomic DNA into small double-stranded pieces and modifying them for compatibility with the sequencing platform.
What are the two types of filaments in a sarcomere?
Thick and thin filaments.
What is the diameter of nanopores?
~ 1 nm.
How do postganglionic neurons differ from typical neurons at the neuro-effector junction?
They don’t have discrete axon terminals.
What is the A band in a sarcomere?
A dark band in the middle of a sarcomere containing thick filaments and overlapping thin filaments.
What is the level of organization that consists of different types of tissues working together?
Organs.
What is a muscle fascicle?
A bundle of muscle fibers (cells).
What role does titin play in muscle structure?
It provides structural support and elasticity to maintain the alignment of thick filaments in the middle of the sarcomere.
What does Sanger Sequencing synthesize?
Partial copies of a target DNA that vary in length by one dideoxynucleotide (ddNTP).
How many myosin molecules compose a thick filament?
300 myosin molecules.
What is isometric muscle contraction?
A type of contraction where muscle length does not change, but tension increases.
What do sympathetic preganglionic neurons connect with in the sympathetic trunk ganglia?
Postganglionic neurons.
What is the connective tissue layer surrounding a muscle fascicle called?
Perimysium.
What energy pathway is used for prolonged-duration exercise?
ATP is generated by breaking down several nutrients through the aerobic pathway.
What is the primary function of skeletal muscle?
Supporting and moving the skeleton.
What initiates signal transmission at neuro-effector junctions?
An action potential arrives at the varicosity.
How does each base affect the current in nanopore sequencing?
Each base blocks the current differently due to its unique structure.
What is the final step in the NGS workflow?
Analyze the data.
What type of control governs heartbeats and digestion?
Involuntary control by the autonomic nervous system.
What issue arises when a read falls into a repeated area of the genome?
It is impossible to determine the exact number of repeats.
What is a varicosity in the context of autonomic neuro-effector junctions?
Swellings along the axons of neurons where neurotransmitters are released.
What is a key feature of adapters in library preparation?
A sequence that is complementary to the oligonucleotides bound to the flow cell or bead.
Which brain structure regulates the medulla oblongata?
Hypothalamus.
What does the head region of myosin contain?
ATP and actin binding sites.
What happens when a voltage is applied across the membrane of a nanopore detector?
Ions flow through the open channel (nanopore).
What is the relationship between muscle, tendon, and bone?
Muscle is attached to bone via tendon.
Why is a skeletal muscle cell referred to as a muscle fiber?
Because of its elongated shape, resembling a fiber.
What binds to the adrenergic receptor on the effector cell?
Norepinephrine (NE).
What structures are found among the muscle fascicles?
Vessels and nerves.
What is another name for the plasma membrane in muscle cells?
Sarcolemma.
What is a key advantage of the third generation of sequencing?
Lower cost compared to previous generations.
What happens during exercise lasting 30 to 40 seconds?
Glycogen stored in muscles is broken down to glucose, which is oxidized to generate ATP (anaerobic pathway).
What is the main function of the Autonomic Nervous System (ANS)?
To innervate organs and tissues under involuntary control.
What are the three regions of a myosin molecule?
Head, Neck, and Tail.
What is the fourth step in the NGS process?
Sequence each cluster.
How does DNA move through the nanopore?
DNA is pulled through the nanopore to the positive side due to its negative charge.
What triggers the release of neurotransmitters from varicosities?
An action potential in the postganglionic neuron.
What does the regulation of neuro-effector transmission depend on?
The concentration of transmitter available for binding.
Is acetylcholine an excitatory or inhibitory neurotransmitter for skeletal muscle?
Excitatory neurotransmitter.
Do thick and thin filaments get shorter during muscle contraction?
No, neither thick nor thin filaments get shorter.
How does a computer decipher the DNA sequence in nanopore sequencing?
By measuring the current changes and differences caused by the bases.
Where is acetylcholine synthesized?
In the axon terminal.
What enzyme is involved in muscle contraction?
Myosin ATPase.
What is the function of the inferior mesenteric ganglion?
It innervates the large intestine and pelvic organs.
Who is the author of the comparison among the three generations of sequencing?
David Cheung.
What proteins can be used to create nanopore detectors?
Proteins such as alpha-hemolysin from Staphylococcus.
How does Ion Torrent sequencing compare to Illumina-based methods in terms of speed?
Ion Torrent sequencing is much faster than Illumina-based methods.
What are the three ways to generate ATP in muscle cells?
Phosphorylation of ADP by creatine phosphate, oxidative phosphorylation of ADP in mitochondria, and anaerobic glycolysis.
What happens when depolarization occurs at the neuro-effector junction?
Voltage-gated Ca2+ channels open, causing an influx of Ca2+.
What are terminal cisternae?
Enlarged regions at the end of each sarcoplasmic reticulum.
In which direction does the tail of myosin point?
Towards the M line region.
What happens to the muscle during contraction according to the sliding filament theory?
The muscle shortens by overlapping more of the thick and thin myofilaments.
What does the celiac ganglion primarily innervate?
It innervates the stomach, liver, spleen, and pancreas.
What happens when sympathetic preganglionic neurons synapse in the ganglion they first reach?
They synapse with postganglionic neurons at the same spinal level.
What type of muscle is primarily responsible for movement in the body?
Skeletal muscle.
Which ganglion is associated with the innervation of the small intestine?
The superior mesenteric ganglion.
How is the DNA sequence determined from the gel?
By reading off the bands in order, starting at the bottom of the gel and reading across all lanes.
What is the pathway of the sympathetic system in pupillary reflexes?
Pre-ganglionic fibers project from the hypothalamus to the superior cervical ganglion, then postganglionic fibers project to the pupillary dilator muscles.
What enzyme is responsible for the synthesis of acetylcholine?
Choline acetyl transferase.
What is another example of Third-Generation Sequencing?
Single-molecule Real-time (SMRT) Sequencing.
What color light does the fluorophore for G emit?
Green light.
What neurotransmitter is secreted by sympathetic postganglionic neurons?
Norepinephrine (NE) or epinephrine (E).
What is the purpose of the oligonucleotides on the flow cell?
They are complementary to the adapter ends of the gDNA fragments.
What is the first step in Next-Generation Sequencing (NGS)?
Genomic DNA (gDNA) isolation.
What is the simplest level of organization in the human body?
Cells.
What is the function of mitochondria in muscle fibers?
Energy support.
What type of muscles does the ANS provide motor control to?
Smooth muscle, cardiac muscle, glands, and adipocytes.
What is the purpose of the index (barcode) sequence in library preparation?
It is unique to the genomic fragments from one sample, used when mixing multiple samples into one reaction.
What is the role of the aorticorenal ganglion?
It innervates the kidneys and adrenal glands.
What happens when there are two identical T nucleotides in a row during Ion Torrent sequencing?
DNA polymerase adds two dATP, leading to a change in pH and voltage.
What is the role of the pretectal nuclei in pupillary reflexes?
They project to the Edinger-Westphal nucleus.
Where does titin anchor thick filaments?
Between the Z line and M line.
What change occurs to the lens when viewing a near object?
The curvature of the lens increases to become convex.
What is each myofilament made of?
1 myosin molecule.
What complex covers the active sites of actin when muscle contraction is turned off?
Troponin-tropomyosin complex.
What are the key zones and lines in a sarcomere?
A band, Z line, I band, H zone, and M line.
What is the reference genome for humans available for viewing?
The human genome at the Genome Reference Consortium Website.
What enzyme is involved in muscle relaxation?
Ca2+ ATPase in the sarcoplasmic reticulum (SR).
Where does the degradation of acetylcholine occur?
In the synaptic cleft.
Where is Ca2+ ATPase located?
In the sarcoplasmic reticulum (SR).
What is the main method used in Illumina Sequencing?
Sequencing by Synthesis.
Why are multiple copies of the genome mixed together in gDNA isolation?
To ensure multiple gDNA fragments from each chromosome in the final gDNA library.
What happens to the circular muscles in bright light?
They contract.
What physiological processes does the ANS regulate?
Heart rate, blood pressure, and digestion.
What is the exception for sympathetic postganglionic neurons?
Sympathetic cholinergic neurons terminate on sweat glands and secrete acetylcholine.
How do Ion Torrent sequencing and Illumina sequencing differ in terms of DNA fragment attachment?
In Ion Torrent sequencing, DNA fragments are attached to microbeads, while in Illumina sequencing, they are attached to a flow cell.
What is considered a sufficient average read depth for whole genome sequencing?
An average read depth of 20-30.
What is the purpose of Emulsion PCR in Ion Torrent Sequencing?
To physically separate the beads and amplify the gDNA attached to the beads.
How many clusters can be found on a single flow cell?
Tens of millions.
Can we voluntarily control our heartbeats?
No, heartbeats are primarily controlled involuntarily by the autonomic nervous system.
What is isotonic muscle contraction?
A type of contraction where tension increases and muscle length changes.
What surrounds the muscle body in skeletal muscle?
Epimysium.
What is the structure of a myosin molecule?
It has 2 subunits twisted around each other to form a dimer.
What level of organization comes after cells in the human body?
Tissues.
What happens when sympathetic preganglionic neurons continue without synapsing?
They end at a prevertebral ganglion and synapse with postganglionic neurons.
What is the significance of striation in skeletal muscle?
It indicates the organized arrangement of myofilaments.
What technique is used to separate the products of the four sequencing reactions?
Electrophoresis in a polyacrylamide gel.
What happens to the ciliary muscles when viewing a near object?
They contract due to parasympathetic stimulation (M3 muscarinic receptors).
What is required for muscle contraction and relaxation?
ATP.
What effect does the contraction of ciliary muscles have on the zonular fibers?
The zonular fibers become slack.
What is a key difference between Third-Generation Sequencing (TGS) and Next-Generation Sequencing (NGS)?
TGS can decode single copies of genomic DNA fragments, while NGS requires multiple copies.
What happens when light falls into one eye?
It stimulates retinal photoreceptors, bipolar cells, and retinal ganglion cells.
What causes receptor activation to cease?
Norepinephrine (NE) diffuses away from the synapse.
What type of muscle does the somatic nervous system excite?
Skeletal muscle.
How is norepinephrine (NE) removed from the synapse?
It can be taken back into the synaptic vesicles for re-release or metabolized by monoamine oxidase (MAO).
What triggers the release of neurotransmitters in autonomic neuro-effector junctions?
An action potential triggers the release from all varicosities.
What type of fibers do pre ganglionic neurons use?
Mildly myelinated B fibers.
What is located at the axon terminal in the neuromuscular junction?
Somatic motor neuron.
How do the parasympathetic axons contribute to pupillary reflexes?
They synapse with ciliary ganglion neurons to innervate the pupillary constrictor muscles.
What enzyme is responsible for the degradation of acetylcholine?
Acetylcholinesterase.
What type of muscle tissue is found in hollow organs?
Smooth muscle.
What are adapters used for in NGS?
Adapters provide binding sites for sequencing primers, PCR primers, and are complementary to the oligonucleotide of the beads for binding.
What is the result of channel opening in nicotinic cholinergic receptors?
Depolarization (excitation) of the postsynaptic cell.
What is bridge amplification?
A type of PCR where the gDNA fragment bends over so both adapters anneal to the surface via oligonucleotides.
What is an autonomic ganglion?
A group of neuron cell bodies in the PNS (outside of CNS).
What is the primary function of skeletal muscle?
To facilitate movement of the skeleton.
What is the main component of a nanopore detector?
Nanopores (channels) in a membrane.
What is one way sympathetic preganglionic neurons can connect with postganglionic neurons?
They can extend to chromaffin cells of the adrenal medulla.
What challenge is faced when aligning reads derived from repeated regions?
It is difficult to determine where the read should be aligned, as it could fit multiple locations in the genome.
What is required for muscle contraction and relaxation?
Energy in the form of ATP.
What is a limitation of Ion Torrent sequencing when dealing with gDNA?
It struggles with multiple identical bases in a row.
What occurs when sympathetic preganglionic neurons ascend or descend?
They synapse with postganglionic neurons at a different spinal level.
What do the bands in each lane of the gel represent?
Fragments that have terminated at a given nucleotide.
What does the reduction in ionic current depend on?
The base sequence of the DNA.
Which parts of the brain also influence the Autonomic Nervous System?
Cerebral cortex and cerebellum.
What is the M line in a sarcomere?
A dark line in the center of the H zone that links thick filaments together.
What technology does SMRT sequencing utilize to visualize DNA polymerase?
Zero-mode waveguides.
What occurs between the thick and thin filaments during contraction?
The filaments slide and pass each other.
What is the first step in the NGS process according to David Cheung?
To separate or partition the library of gDNA fragments into discrete locations on a solid surface.
What is the purpose of the fluorescent tag in SMRT sequencing?
It emits light upon release from the DNA to identify nucleotide addition.
What role does sympathetic stimulation play when viewing a distant object?
It increases via β2-adrenergic receptors.
What type of receptors are nicotinic cholinergic receptors?
Ionotropic receptors associated with ion channels.
What is the role of dideoxyribonucleoside triphosphates (ddNTPs) in the Sanger method?
They terminate the DNA chain during replication.
What are the two main types of adrenergic receptors?
α (alpha) and β (beta) receptors.
What ions can move through nicotinic cholinergic receptors?
Both Na+ and K+ ions.
What change occurs to the lens when viewing a distant object?
The curvature of the lens becomes less convex.
What is the Z line in a sarcomere?
A network of interconnecting proteins attached to one end of thin filaments.
What is the role of actin in muscle contraction?
Actin serves as a contractile protein with myosin binding sites.
What is a limitation of nanopore sequencing technology?
It suffers from inaccuracy that often leads to sequencing errors.
What is one medical application of botulinum toxin?
To relieve overactive muscle movements.
What is the typical read length for SMRT Sequencing?
20,000 bases.
What cosmetic benefit does botulinum toxin provide?
Reduces face wrinkles and lines.
What distinguishes cardiac muscle from other muscle types?
It is involuntary and striated, found only in the heart.
What is done with excess dGTP after data recording?
It is removed from the chip.
What is the role of the transverse tubule (T-tubule) in muscle contraction?
It transduces the membrane depolarization (action potential) to the muscle fiber.
What is a limitation of Next-Generation Sequencing (NGS) related to the human genome?
It arises from the repetitive nature of the human genome.
What is one function of skeletal muscle?
Posture maintenance.
What is another function of skeletal muscle?
Movement.
What part of the brain is the center for the control of body temperature, hunger, and thirst?
Hypothalamus.
What happens to the current as ssDNA transits the nanopore?
The detector records how the current changes.
What is titin?
An elastic protein that anchors thick filaments between the Z line and M line.
What is Sanger Sequencing also known as?
Chain Termination Sequencing.
What is the mechanism of muscle contraction known as?
Sliding filament theory.
What is the primary neurotransmitter found in the neuromuscular junction?
Acetylcholine.
What does the adapter sequence need to be complementary to for successful sequencing?
The sequencing primer.
What level of organization is formed by groups of organs that work together?
Organ systems.
What is the function of the Na+/K+ ATPase pump?
It establishes resting membrane potential.
What role does the Na+/K+ ATPase pump play in muscle cells?
It maintains the resting membrane potential.
What is a key advantage of nanopore sequencing technology?
Its high speed and ability to handle long DNA molecules.
What is the role of the limbic system in relation to the ANS?
Responsible for visceral responses that reflect emotional states.
What is used to hybridize with the DNA in the Sanger method?
A short DNA primer labeled with a fluorescent dye or radioisotope.
What defines one sarcomere?
The area between two Z lines (Z disks).
What does the sequence obtained from the gel represent?
It is complementary to the sequence of the original gray single-stranded DNA.
What device do Illumina sequencing platforms use for partitioning?
A flow cell.
What is botulinum toxin commonly known as?
Botox.
How do neurotransmitters diffuse in autonomic neuro-effector junctions compared to synaptic clefts?
Neurotransmitters diffuse over a greater area of the effector organ in neuro-effector junctions.
How is the base recorded when the fragment emits light?
The detector records the color and reports it as the corresponding base (e.g., G or A).
What type of receptors do target cells have for norepinephrine and epinephrine?
Adrenergic receptors.
Which neurotransmitters are associated with the sympathetic nervous system?
Epinephrine and Norepinephrine.
How does SMRT sequencing read the DNA sequence in real time?
By identifying the nucleotide base bound to pyrophosphate as it is added to the growing strand.
What occurs when contraction is turned off in terms of actin and myosin?
Actin and myosin can't bind with each other.
What are the two branches of the Autonomic Nervous System?
Sympathetic and parasympathetic divisions.
What is released when dGTP is added, and what does it affect?
A hydrogen ion (H+) is released, changing the pH of the liquid in the well.
Which neurotransmitter has a higher affinity for α1 and α2 receptors?
Norepinephrine (NE) has a higher affinity than epinephrine (Epi).
How does the read length of SMRT Sequencing compare to Illumina reads?
SMRT reads are significantly longer at 20,000 bases compared to Illumina reads, which are between 100 and 200 bases.
What is a 'read' in the context of Illumina Sequencing?
The final sequence for each cluster.
How does the ANS contribute to homeostasis?
By regulating organs to maintain homeostasis.
What is the order of base current reduction in nanopore sequencing?
G > C > T > A.
What advantage do nanopores provide in DNA sequencing?
Many long fragments of DNA can be determined simultaneously.
What does the Sanger method produce?
Four sets of labeled DNA molecules.
What is a myofibril?
A basic rod-like organelle of a muscle cell that contains many chains of parallel myofibrils.
How can reads from gDNA be aligned into continuous sequence information?
By comparison to a previously sequenced genome or by comparing overlapping sequences.
What happens to the ciliary muscles when viewing a distant object?
They relax due to decrease/absence of parasympathetic stimulation.
What happens to the DNA fragments as they pass through the laser?
The fluorophore at the 3' end emits a characteristic color of light for the base.
What is the primary neurotransmitter of the parasympathetic nervous system?
Acetylcholine (Ach).
What are the two types of myofilaments?
Thick filament (mainly composed of myosin protein) and thin filament (mainly composed of actin protein).
What happens to the H zone during muscle contraction?
It decreases in size.
What happens to the zonular fibers when viewing a distant object?
They tighten and pull tight on the lens.
What is the function of the Z line in a sarcomere?
It serves as a network of interconnecting proteins attached to the ends of thin filaments.
How does the Sanger method determine the complete sequence of DNA?
By producing DNA copies that terminate at different points in the sequence.
What is the benefit of assembling many nanopores in a small region?
Many long fragments of DNA sequence can be decoded simultaneously.
What is the significance of read depth in sequencing?
Higher read depth increases confidence in identifying correct bases.
What happens to the radial muscles in dim light?
They contract.
How many subtypes are there for β adrenergic receptors?
Three subtypes: β1, β2, and β3.
What does each cluster on the flow cell represent?
One small segment of the whole genome.
What is the role of the sympathetic division?
It prepares the body for 'fight or flight' responses.
What is the primary feature of smooth muscle?
It is involuntary and controls the movement of hollow organs.
What condition can botulinum toxin treat related to sweating?
Hyperhidrosis (excessive sweating).
What does grey matter in the CNS contain?
Cell bodies and dendrites of neurons.
What components are mixed into the emulsion during Emulsion PCR?
DNA polymerase, dNTPs, and PCR primers complementary to the adapter.
Why do the PCR components partition into the water droplets?
Because they are water soluble.
What is one method used to shear DNA during library preparation?
Ultrasonic disruption (Sonication).
What systems does the medulla oblongata control?
Cardiovascular, pulmonary, urinary, reproductive, and digestive systems.
What does the H zone represent in a sarcomere?
A light band in the middle of the A band, containing thick filaments only with no overlap.
What is the role of the synapse in autonomic neuro-effector junctions?
It connects an efferent neuron to its effector organ.
What is a key structural difference between autonomic neuro-effector junctions and synaptic clefts?
Autonomic neuro-effector junctions involve varicosities, while synaptic clefts involve axon terminals.
What is the highest level of organization in the human body?
Organism.
What does Myosin ATPase do?
It provides energy for muscle contraction.
How does the size of autonomic neuro-effector junctions compare to synaptic clefts?
Autonomic neuro-effector junctions are bigger, while synaptic clefts are smaller.
What are the two types of neurons in the Autonomic Nervous System (ANS)?
Pre ganglionic neurons and post ganglionic neurons.
What happens to the size of the A band during muscle contraction?
It remains the same.
What are the four normal deoxyribonucleoside triphosphates (dNTPs) used in the Sanger method?
A, C, G, and T.
What are zero-mode waveguides?
Small nanosized metal cylindrical wells that reduce background light for visualization.
What is the first step in Ion Torrent sequencing?
Fragmenting the gDNA sequence into millions of tiny double-stranded pieces (~300 – 800 bp).
What is the response time of the somatic nervous system?
Faster response time.
What is the motor end plate?
The plasma membrane of the muscle fiber that lies under the axon terminal.
What is the mechanism of neurotransmitter action in both neuro-effector junctions and synaptic clefts?
Both involve ligand (neurotransmitter) receptor binding.
What is the basic unit of muscle contraction?
Sarcomere.
What initiates the mechanism of muscle contraction?
An action potential reaching the axon terminal of a motor neuron.
What are the receptor types for norepinephrine?
α and β receptors.
What type of receptors are muscarinic cholinergic receptors?
Metabotropic receptors.
What special structure do sympathetic ganglia form?
Sympathetic trunk, which runs parallel to the spinal column.
How does troponin regulate muscle contraction?
Troponin binds to Ca2+ and moves the tropomyosin complex to expose the myosin binding site on actin.
What is the affinity of β1 receptors for norepinephrine and epinephrine?
Norepinephrine (NE) and epinephrine (Epi) have equal affinity.
What does Emulsion PCR prevent?
Cross-contamination of PCR reactions between different beads.
What occurs after the burst of color in SMRT Sequencing?
The fluorophore is washed away, and the next cycle begins.
What is a cosmetic use of botulinum toxin?
Smoothing the skin.
What happens to each bead after multiple PCR cycles?
Each bead becomes coated with thousands of copies of the original gDNA fragment.
What types of cells are found in the adrenal medulla?
Modified sympathetic postganglionic cells called chromaffin cells that develop into endocrine cells.
What is a sequencing library?
A pool of DNA fragments with adapters attached.
What is the difference between the motor control of the somatic nervous system and the autonomic nervous system?
The somatic nervous system innervates skeletal muscles for deliberate motion, while the ANS innervates smooth and cardiac muscles for involuntary motion.
What is the role of transverse tubules (T-tubules) in muscle fibers?
They are tubular structures that are continuous with the plasma membrane and allow action potentials to propagate inside the muscle cell.
How can the bands in the gel be visualized?
With X-ray film (if labeled with radioactive isotope) or UV lamp (if labeled with fluorescence).
What factors affect the release of norepinephrine (NE) from sympathetic neuron axons?
Frequency of action potential at the varicosity, amount of NE available for release, and other modulating factors.
What technique is used to separate copies of target DNA in the provided text?
Capillary Electrophoresis.
What is one example of Third-Generation Sequencing?
Nanopore Sequencing.
What are myofilaments?
The smaller parallel fiber structures that make up a myofibril.
What type of receptors do postganglionic cells have for acetylcholine?
Nicotinic cholinergic receptors (AChR).
What is a notable feature of current nanopore detectors?
They are small enough to be carried everywhere.
Where are α-adrenergic receptors typically found?
In the presynaptic membrane.
What is the significance of the green arrow in the context of the gel?
It indicates the DNA sequence of the newly synthesized strand.
What type of fibers do post ganglionic neurons use?
Unmyelinated C fibers.
What is the impact of agonists and antagonists on NE release?
They modulate NE release.
What are ZMWs in SMRT Sequencing?
ZMWs are nanocontainers where reactions are carried out, reducing background light for detecting individual nucleotide flashes.
How many subtypes are there for α adrenergic receptors?
Two subtypes: α1 and α2.
What role does the neuromuscular junction play in muscle contraction?
It facilitates communication between the neuron and muscle fiber to initiate contraction.
How many sarcomere units does one myofibril have?
Many sarcomere units.
What neurotransmitter is secreted by parasympathetic postganglionic neurons?
Acetylcholine (ACh).
What is the response time of the autonomic nervous system?
Slower response time.
What type of receptors do target cells have for acetylcholine in the parasympathetic system?
Muscarinic cholinergic receptors.
What type of receptor does acetylcholine bind to at the neuromuscular junction?
Nicotinic acetylcholine receptor.
What neurotransmitter do postganglionic neurons of the parasympathetic nervous system primarily release?
Acetylcholine.
What triggers muscle contraction at the molecular level?
High Ca2+ concentration causes Ca2+ to bind to troponin, exposing the myosin binding site on actin.
What must gDNA be in order to proceed with library preparation?
Pure and contain only the DNA portion of the sample.
Where is the ryanodine receptor (RyR) located?
On the sarcoplasmic reticulum (SR) membrane.
What is the effect of catecholamines released into the bloodstream?
They cause widespread effects of sympathetic activation.
What is the primary function of the parasympathetic division?
Rest and digest.
What occurs when there are more than three identical nucleotides in a row in Ion Torrent sequencing?
The pH and voltage changes do not vary significantly, leading to potential sequencing errors.
How are the synthesized DNA fragments separated in Sanger Sequencing?
By size.
What neurotransmitter is released by both sympathetic and parasympathetic preganglionic neurons?
Acetylcholine (ACh).
What role do auto receptors play in NE release?
They provide negative feedback control to conserve and stabilize NE concentration within the synaptic cleft.
What are the three types of muscle tissues?
Skeletal muscle, smooth muscle, and cardiac muscle.
What is the role of Ca2+ ATPase in muscle contraction?
It removes Ca2+ from the cytosol and transports it from the cytosol to the sarcoplasmic reticulum (SR).
What is the I band in a sarcomere?
A light band between the A bands of two sarcomeres, containing only thin filaments with no overlap.
Which type of muscle tissue is under voluntary control?
Skeletal muscle.
How do nanopore devices connect to computers?
Via a typical USB port.
What happens to the I band during muscle contraction?
It decreases in size.
What happens to Ca2+ ions when muscle contraction is turned off?
Ca2+ ions leave troponin.
What components are supplied to determine the sequence of gDNA fragments?
A sequencing primer, DNA polymerase, and fluorescently-labeled nucleotides.
How does botulinum toxin function at the neuromuscular junction?
It inhibits acetylcholine release.
What is the axon length of sympathetic preganglionic neurons?
Short.
What format are the final results compiled into for analysis?
Sequence files in the form of a chromatogram.
In what scenarios does nanopore technology provide accurate enough data?
For resequencing or for quick analysis.
What is the significance of different fluorophores for each base?
Each base has a different fluorophore that emits a unique color, allowing for identification.
How do sympathetic preganglionic neurons relate to postganglionic neurons?
They form synapses at the same level or send collaterals to other spinal levels.
What does a voltage change recorded as a 1 or 0 signify?
A 1 means the nucleotide was added, and a 0 means it was not added.
What are the two main divisions of the nervous system?
Central nervous system and peripheral nervous system.
How many reads can be generated in a single flow cell?
Tens of millions of reads.
Are adrenergic receptors homogeneously distributed throughout tissues?
No, they are not homogeneously distributed.
What is the role of long preganglionic neurons in the SNS?
They innervate the adrenal medulla of the adrenal gland without synapsing with postganglionic neurons.
What catecholamines are released by the adrenal medulla upon stimulation?
80% epinephrine, 20% norepinephrine, and a small amount of dopamine.
What are some effects of sympathetic activation?
Raising heart rate, constricting blood vessels, and releasing glucose.
What is the consequence of misinterpreting pH/voltage changes in Ion Torrent sequencing?
It can lead to sequencing errors.
How is the resulting sequence processed in nanopore sequencing?
The sequence is listed in real-time as data are processed.
What is the neuromuscular junction?
The junction between the axon terminal and the motor end plate.
Which muscles are innervated by the sympathetic system for pupillary dilation?
Pupillary dilator muscles (radial muscles).
What is de novo sequencing?
Sequencing DNA from a species that does not have a reference genome using alignment of overlapping sequences.
What is the first step in Ion Torrent Sequencing after emulsion PCR?
The oil is removed and each bead is partitioned into separate microwells on a semiconductor chip.
What is a sarcomere?
The functional unit of myofibril that contains thick and thin filaments and forms a repeating pattern.
What is the process of actin polymerization?
Actin monomers polymerize to form two strands that create a double helix.
Where do preganglionic neurons of the sympathetic nervous system originate?
From the lateral horn of the thoracic and lumbar spinal cord (T1 - L2).
What color light does the final nucleotide emit in the example provided?
Yellow light.
What happens when a specific dNTP, like dGTP, is added to the chip?
If a well contains a gDNA fragment with a C next to the primer, dGTP is added; otherwise, it is not.
What role does the fluorophore play in the sequencing process?
It acts as a blocking group, preventing DNA polymerase from adding another nucleotide until removed.
What activates voltage-gated Ca2+ channels during muscle contraction?
Membrane depolarization.
What type of muscle does acetylcholine affect?
Skeletal and smooth muscle.
What are muscarinic cholinergic receptors coupled with?
G-proteins (Guanine nucleotide-binding proteins).
What is the role of the parasympathetic division?
It promotes 'rest and digest' functions.
What happens to troponin and tropomyosin during muscle relaxation?
At low Ca2+ concentration, the complex blocks the actin-myosin binding site, leading to muscle relaxation.
What effect does opening Na+/Ca2+ channels through muscarinic receptors have?
Depolarization and excitation (contraction of smooth muscles in the GI tract).
What does white matter in the CNS consist of?
Tracts of myelinated axons.
How do parasympathetic preganglionic neurons relate to postganglionic neurons?
They form synapses with short postganglionic neurons.
What processes are allowed by the release of Ca2+ during muscle contraction?
Cross-bridge cycle and filament sliding in the sarcomere, leading to muscle contraction.
What is the role of dual innervation in visceral organ control?
It ensures precise control over the activity of the visceral organ.
How many neurons connect the CNS to the organ in the somatic nervous system?
1 neuron (monosynaptic).
What are the three major proteins of thin filaments?
Actin, Tropomyosin, and Troponin.
What are the two types of receptors for acetylcholine?
Nicotinic and Muscarinic.
What is the Autonomic Nervous System (ANS)?
A division of the peripheral nervous system that regulates involuntary physiological processes.
What type of protein is botulinum toxin?
A neurotoxic protein from the bacteria Clostridium botulinum.
What happens after the gDNA fragments attach to the flow cell?
PCR amplification occurs to create a cluster.
What is the role of neurotransmitters in the ANS?
They can be excitatory or inhibitory depending on the receptors.
Which muscle tissue is involuntary and found in the heart?
Cardiac muscle.
How many neurons are needed to connect the CNS to the organ in the autonomic nervous system?
2 neurons (di-synaptic).
What does tropomyosin do in muscle fibers?
Tropomyosin covers the active site on actin to prevent actin-myosin binding.
How does DNA polymerase add nucleotides during sequencing?
It adds the complementary fluorescently-labeled nucleotide to the gDNA and connects it to the end of the primer.
How do semiconductor chips detect nucleotide addition?
They have miniature pH meters that measure the release of H+ during phosphodiester bond formation.
What are the components of the CNS?
Brain and spinal cord.
Where do preganglionic neurons of the parasympathetic nervous system originate?
From brainstem nuclei (CN III, VII, IX, X) and the sacral spinal cord (S2 - S4).
What is the first step in library preparation for NGS?
Isolate gDNA from a sample (e.g., tissue or blood).
What is the process repeated until the gDNA has been sequenced?
Adding remaining dNTPs, recording data, and washing away excess dNTPs.
What is the somatic nervous system responsible for?
Voluntary control of skeletal muscles.
What is the purpose of using Tn5 transposase in library preparation?
To cut chromosomes into small fragments and tag the ends with adapters.
How do zero-mode waveguides enhance the visualization process in SMRT sequencing?
They allow only a small portion of the cylinder to be visualized for fluorescent light flashes.
What is added to each well during the sequencing process?
DNA polymerase and a sequencing primer.
What type of muscle does the autonomic nervous system inhibit or excite?
Smooth muscle, cardiac muscle, or glandular secretion.
What happens to the DNA during the Sanger sequencing process?
It is divided into four reaction tubes, each receiving a different ddNTP.
How does SMRT Sequencing avoid accumulating tags on DNA?
By attaching the fluorophore to the discarded pyrophosphate group instead of the incoming nucleotide.
What is the length of the DNA template that can be used in SMRT sequencing?
Tens of thousands of bases in length.
What is the significance of the microbeads in Ion Torrent sequencing?
Each microbead is coated with a complementary oligonucleotide to the fragment’s adapter sequence, allowing for the attachment of only one DNA fragment per bead.
What triggers the exocytosis of acetylcholine-containing vesicles?
Ca2+ influx.
What neurotransmitter do postganglionic neurons of the sympathetic nervous system primarily release?
Norepinephrine.
What effect does opening K+ channels through muscarinic receptors have?
Hyperpolarization and inhibition (decrease heart rate).
What happens when the nicotinic acetylcholine receptor is activated?
It opens a ligand-gated Na+ ion channel, causing Na+ influx.
What is the affinity of β3 receptors for norepinephrine and epinephrine?
Norepinephrine (NE) and epinephrine (Epi) have equal affinity.
What does the dihydropyridine receptor (DHP receptor) do?
Senses the action potential on the T-tubule membrane and relays information to the ryanodine receptor (RyR).
What does ultrasonic disruption do to the DNA?
Breaks the molecular bonds holding the DNA backbone together.
What is Tagmentation?
The process of cutting DNA and adding adapters using Tn5 transposase.
How does the somatic nervous system differ from the autonomic nervous system?
The somatic nervous system has one somatic motor neuron that extends from the CNS to skeletal muscle fibers and is always excitatory.
How are beads separated during Emulsion PCR?
By encapsulating them in small drops of liquid suspended in a solution of oil.
What type of information does each cluster provide?
A small length of DNA sequence information.
Which neurotransmitter has a significantly higher affinity for β2 receptors?
Epinephrine (Epi) has a much higher affinity than norepinephrine (NE).
What is the axon length of parasympathetic preganglionic neurons?
Long.
How does the computer identify the added nucleotide?
By the color of its fluorescence.
What is released when the ryanodine receptor (RyR) opens?
Ca2+ ions.
How do the sympathetic and parasympathetic divisions typically interact?
They are usually activated reciprocally; when one division's activity increases, the other's decreases.
What is the primary function of the sympathetic division?
Fight or flight response.
What happens after the fluorescence of the added nucleotide is recorded?
The fluorophore is removed, allowing DNA polymerase to add another complementary fluorescently-labeled nucleotide.
What happens during each extension reaction in SMRT Sequencing?
Each extension reaction gives a brief burst of color, indicating nucleotide incorporation.
What effect does botulinum toxin have on muscle contraction?
It weakens muscle contraction.
What does the sensory (afferent) division do?
Transmits sensory information to the central nervous system.
What does the motor (efferent) division do?
Transmits motor commands from the central nervous system to the body.
What are collateral ganglia?
Ganglia located within the abdominal cavity, such as aorticorenal, celiac, superior mesenteric, and inferior mesenteric ganglia, that are closer to the innervated organs.
Which organs receive only sympathetic nerve input?
Most blood vessels, sweat glands, and the adrenal medulla.
What are some effects of parasympathetic activation?
Digesting food, expelling waste, and general maintenance.
What types of cells are affected by epinephrine and norepinephrine?
Cardiac muscle, smooth muscle, and gland cells.
Where do parasympathetic ganglia typically terminate?
Near or within the effector organs.
What are the sympathetic trunks?
They are paravertebral ganglia that lie close to the spinal cord and form two chains of ganglia, one on each side of the cord.
Why are adapters added to the ends of DNA fragments?
To provide a place for sequencing primers to anneal and to attach to the flow cell.
What is dual innervation in the autonomic nervous system?
Both divisions of the ANS innervate most effector organs and function antagonistically.
