Lesson 10 - Learning & Memory; Neuroregeneration

Learning and Memory

Learning Objectives:
● Describe the different stages of memory processing
○ acquisition, retention, consolidation, retrieval
● Describe and distinguish between the different forms of memory
○ declarative v.s. procedural
○ short-term v.s. long-term v.s. working memory
● Outline the brain structures involved in different forms of memory processing
○ temporal lobe and declarative memory
○ striatum and procedural memory
○ hippocampus and spatial memory
○ prefrontal cortex and working memory
● Discuss the concept of the Hebbian theory of synaptic plasticity
● Describe the processes involved in long-term potentiation and its association with memory

Learning, Memory and Intelligence

● Learning - the process by which we acquire knowledge about the world
● Memory - the process by which knowledge of the world is encoded, stored and later
retrieved
● Intelligence - a very general mental capability that involves the ability to reason, solve
problems, think abstractly, comprehend complex ideas, and learn quickly and from
experience.

Correlation between cortical volume and intelligence

● There is a positive correlation between overall brain size & intelligence test scores. The
bigger the brain, the smarter you are. The more complex the connections are, the better
the ability of the brain to process information.
● This is particularly true in the prefrontal and temporal cortices, regions responsible for
processing cognitive and executive functions. Also, the prefrontal and temporal cortices
have the hippocampus, an important region for learning and memory

*Males have a slight but consistently wider distribution than females at both ends of the range
Different types of memory
● Short-term memory
○ Memories that last seconds to hours
○ Vulnerable to disruption and readily lost (needs to be consolidated)
○ Working memory
■ Temporary form of information storage
■ Limited in capacity and requires rehearsal
■ Retention of a telephone number that has just been given to you by
repetition
● Long-term memory
○ Converted from short-term memory by consolidation
○ Lasts longer with re-consolidation (e.g. remembering your parents’/siblings’
birthdays)
○ Can last for years (e.g. recall of childhood memories)
○ Declarative/Explicit memory
■ Memory with conscious recall
■ Episodic – autobiographical information with temporal or spatial context.
● Memories of conscious events in our lives that have a specific source
in time, space, and circumstances (e.g., your first day of school or
your first kiss)
■ Semantic – memory for facts, general knowledge or events with no
associations with you, you don’t know why you remember (e.g., Kyoto is a
city in Japan)
○ Non-declarative/Implicit memory
■ Memory without conscious recall
■ Procedural – memory for skills and habits (e.g., using chopsticks or riding a
bike → muscle memory)
■ Classical conditioning – emotional responses
■ Non-associative – habitation, sensitisation (e.g., going to an exam after a
few practises)

Brain structures involved in different forms of memory processing

● Temporal lobe & Declarative memory
○ Temporal lobe
■ Medial temporal lobes, located towards the ventral surface, include the
hippocampus (comprising the dentate gyrus and subiculum) and adjacent
perirhinal, entorhinal, and parahippocampal cortices.
➢ Hippocampus
➢ Subiculum
➢ Parahippocampus
➢ Rhinal Cortical Area
■ Responsible for processing declarative memory (facts and events we can
consciously recall).
■ Together with the neocortex, it establishes, maintains, and enables the
storage of long-term memory through a process called consolidation.
○ Effects of Temporal Lobectomy (epileptic subject HM)
■ Removal of temporal lobes had no effect on perception, intelligence,
personality
■ Caused profound anterograde amnesia (an inability to form new memories)
and partial retrograde amnesia (inability to recall past events)
➢ So profound that one cannot perform basic human activities
➢ Patients did not recognise Brenda Milner, who had studied him for
nearly 50 years
 ■ Impaired declarative memory but spared procedural and working memory
➢ Intact short-term and procedural memory suggested that the
hippocampus is not crucial for these types of memories.
➢ The hippocampus is also not important for encoding or retrieval of
long-term memories that existed before his lobectomy
● Striatum & Procedural memory
○ Procedural Memory
■ Type of long-term memory of how to perform different actions and
■ Involves the development of motor skills.
■ From very early in life, as you begin to learn how to walk, talk, eat and play,
so ingrained that they are almost automatic.
■ Patients with Parkinson’s disease have impaired procedural memory.
○ Striatum (part of the basal ganglia) is important
■ Consists of caudate nucleus and putamen
■ Prefrontal, parietal cortices and cerebellum are also important for motor
learning.
● Hippocampus & Spatial memory
○ Spatial Memory
■ Part of memory that responsible for recording information about one's
environment and its spatial orientation
■ Important for navigating around a familiar city
○ Hippocampus is important for spatial memory
■ Place cells in the hippocampus - neurons that only fire when a person is at
a particular location
■ Grid cells in the entorhinal cortex - the neurons activate in a unique spatial
pattern in a hexagonal grid
● Prefrontal cortex & Working memory
○ Working Memory
■ Limited capacity system that allows one to temporarily store and process
information over a brief period.
■ A system for temporary storage of information enables one to work on
complex tasks while being able to keep information in mind.
➢ E.g. the ability to work on a complicated mathematical problem
utilizes one’s working memory.
○ Prefrontal cortex
■ Highly developed in humans
■ Important in problem-solving, complex planning and self-awareness
■ Interconnected with the medial temporal lobe

Does Photographic Memory Exist?

● Photographic memory is the ability to remember and recall large volumes of numbers or
text in great detail.
○ There is no scientific evidence that this type of memory exists.
● Eidetic memory is the ability to faithfully recall or reproduce a visual or mental image that
is captured. This form of memory is subject to distortions.
○ Most individuals identified as having eidetic memory are children and prevalence
ranges from 2 – 10 % with no gender difference.

Different Stages of Memory Formation

● Acquisition
 ○ Sensory information is perceived and acquired
○ Influenced by attention, motivation, ability to learn
● Retention/Encoding
○ Crucial first step to creating a new memory
○ The perceived item of interest to be converted into a construct that can be stored
within the brain
○ Require consolidation to commit to longer-term memory
● Consolidation
○ Stabilisation of memory trace after acquisition
○ Two specific processes:
1. Synaptic consolidation - occurs within the first few hours after learning or
encoding
2. System consolidation - where hippocampus-dependent memories become
independent of the hippocampus over a period of weeks to years
○ Utilizes a phenomenon called long-term potentiation, which allows a synapse to
increase in strength.
● Retrieval
○ Subsequent re-accessing of events or information from the past, which have been
previously encoded and stored in the brain.

Hebbian Theory
● States that memory results from synaptic modification
○ If synapses are more closely associated (i.e. wire together), they are more likely to
fire.
○ If they are out of sync (i.e. not firing together), there is a degradation of the
synapses (i.e. lose their link) → If the neurons are not being used constantly, they
are eventually lost.
● It is a neuroscientific theory claiming that an increase in synaptic efficacy arises from
post-synaptic cell’s repeated and persistent stimulation of the post-synaptic cell

Long-term potentiation (LTP) in the Long-term depression (LTD) in the

hippocampus hippocampus

● LTP is induced by tetanic high ● Facilitation of postsynaptic Ca2+ entry

frequency stimulation through NMDA receptors
● Activation of AMPA/Kainate receptors ● Weak depolarization of postsynaptic
by glutamate causes post-synaptic neurons, partial blocking of NMDA
membrane depolarisation channels by Mg2+, trickle of Ca2+ entry
● Mg2+ is dissociated from NMDA ● Activation of protein phosphatases
channels, facilitation of Ca2+ entry instead of kinases
● Activation of protein kinases ● REsulting in protein dephosphorylation
○ Ca2+ /calmodulin-dependent and internalization of AMPA R.
kinase (CaMKII)
○ Ca2+ /phospholipid-dependent
kinase (PKC)
● Resulting in postsynaptic protein
phosphorylation of AMPA receptors
● Recruitment of AMPA receptors to
post-synaptic membrane and
upregulation of AMPA receptor function
 Key difference between LTP and LTD – level of NMDA receptor activation

Dendritic Spines
● Dendritic spines are sources of synaptic contact that can be altered by experience
○ The more dendritic spines there are, the greater the density and thus
synapses, leading to a stronger connection
● Dendritic spines are heterogeneous structures and can readily change the density of
morphology depending on the level of activation of the neuronal networks
● Commonly classified into three types:
○ Thin spines
○ Stubby spines
○ Mushroom spines.
● Structural plasticity of dendritic spines underlies learning, memory and cognition in the
cerebral cortex
● Induction of LTP causes enlargement of spine heads
● Induction of LTD causes spine head shrinkage
Neurodegenerative diseases
● Most have unknown etiology/causes
● Delayed onset; age is a major risk factor
● Selective neuronal vulnerability, despite widespread expression of disease-related proteins
during the whole lifetime
● Abnormal protein processing and aggregation
● Many display symptoms of memory loss
● One of the most common form is Alzheimer’s disease (AD)

Prevalence of dementia
● An estimated 459,000 Australians currently live with dementia with one in three people over
the age of 85 affected
● Dementia is the second leading cause of death in Australia, after heart disease
● The most common condition resulting in dementia is AD

Neuroregeneration and Repair

Learning Objectives:
● Define and name the types of regeneration in the brain
○ Axogenesis
○ Neurogenesis
○ Rewiring
● Describe the differences between peripheral and central nerve regeneration
● Define the classes of stem cell and describe their distinct characteristics
● Discuss the routes to induce pluripotency, namely reprogramming
● Discuss the potential of using stem cells to treat neurodegenerative disorders and restore
brain function
Regeneration
● Regeneration is the replacement of destroyed tissue by the same kind of cells

● Axon regrowth from injured neurons in the peripheral nervous

system through the injury site to re-establish connections
● Regenerative capacity of severed peripheral axons is dependent on
○ Age of subject,
○ Nerve trunk affected,
○ Site and type of lesions,
Axogenesis ○ Type and delay of surgical repair, distance axon must grow to
○ Span injury
○ E.g. Sciatic nerve injury – 10-30% loss of DRG, 50-80%
motor neurons cell death
● Mammalian CNS axons tend not to reinnervate because of factors
secreted by oligodendroglia cells that inhibit growth.
● Nogo which is released when oligodendroglia cells are damaged.

● The production of new neuronal cells from precursor

populations that subsequently produce neurites to make
connections with host cells
● Two areas where neurogenesis occurs is in the subventricular zone
(SVZ) and subgranular zone (SGZ) – maintaining a pool of
self-renewing cells.
Neurogenesis ● The most active area of neurogenesis (occur mostly) is the
hippocampus
 ○ Thousands of new cells are produced in the hippocampus
each day, although many die within weeks of their birth. Some
of these new neurons survive and integrate themselves into
the working brain.
○ The more the animal learned, the more neurons survive
in the hippocampus

● New connections are formed to replace ones that are damaged

to re-establish function of the neural pathway
● This is seen in the young ferret brain.
○ Normally, the visual signals of the LGN and to the visual
Rewiring
cortex while the auditory signals synapses to the MGN
auditory cortex.
○ If the visual signal is lost, the ferret has the ability to rewire
itself, to use the MGN as a relay station to relay visual
information from the retinal ganglion cells form synapses with
the MGN.
○ This does not typically occur in adult humans
● Regeneration of the central nervous system (CNS) requires progenitor cells to migrate,
re-establish cell contacts, self-renew and undergo specification and spatial patterning
to form neurons and glia that will integrate into the host tissue to replace the damaged
structure.
○ Enables newly generated neurons form neurons and glial cells that integrate into
the host tissue to replace damaged structures.
● Understanding the brain’s ability to heal itself or restore function following injury is crucial
due to the increasing prevalence of neurodegenerative diseases and traumatic brain
injuries.
● Sources of cells that can be harnessed for brain repair/regeneration:
1. Enhancing neurogenesis
■ Induction of endogenous neural stem cells to generate new neurons by
stimulating withdrawal neurogenic factors
■ E.g. Boosting the generation of neurons in the hippocampal senate gyrus
through neurogenic compounds
2. Reprogramming neural cells into neurons
■ Ectopic generation of new neurons outside neurogenic niches by
reprogramming neural progenitor cells into new neurons.
■ E.g. By transgenic expression of transcription factors such as
Pax6/Ascl1/Brn2
3. Transplantation to replace injured tissue
■ Transplantation of neural progenitor cells to replace the lost tissue, where
the aim is either to transplant stem cells or the differentiated progeny to
enhanced brain repair.
■ E.g. Transplanting neural precussors to replace dopaminergic neurons

Stem cells hierarchy

● Totipotent stem cells
○ Cells can give rise to all embryonic somatic cells & germ cells.
■ They can build a whole animal.
■ Includes zygote and a few early cells of the morula
● Pluripotent stem cells
○ Descendants of totipotent stem cells
○ Proliferate infinitely & can differentiate into cells of 3 germ layers: ectoderm -
nervous system and skin; mesoderm - musculoskeletal, vascular and lymphatic
systems, kidney and gonads; endoderm - gastrointestinal tract, glands, lung, liver,
pancreas
■ Includes blastocysts
○ 3 sources of pluripotent stem cells:
■ ES – embryonic stem cells from inner cell mass of blastocyst
■ EG – embryonic germ cells from primordial germ cells
■ gPS – germ-line derived pluripotent stem cells from spermatogonical stem
cells of neonatal and adult testis
○ 3 major routes of somatic cell reprogramming to pluripotency
■ Fusion of somatic and ES cells
➢ Involves generating reprogrammed somatic cell through cell fusion
between somatic and pluripotent embryonic stem cells.

■ Nuclear transfer ES cells

➢ Involves transfer of a somatic donor cell into an unfertilized,
nucleated oocyte. The reconstructed cell is stimulated to initiate
development of an embryo

■ Induced pluripotent stem cells (iPSCs)

➢ Adult somatic cells are reprogrammed to acquire similar
characteristics as embryonic stem cells. This is achieved by
introducing certain genes into the cells and transforming them into
embryonic stem cell-like cells. The initial programming required
retroviral transfer into these skin cells of four transcription factors:
Sox2, Oct4, Klf4, and Myc.
 ● Multipotent stem cells
○ Can give rise to cells of a particular lineage or closely related family.
■ Includes haematopoietic cells

Parkinson’s disease
● A neurodegenerative disease involving degeneration of the nigrostriatal dopaminergic
pathway (highly susceptible)
● Debilitating disorder with motor disturbances including tremor, rigidity, bradykinesia and
postural instability
● No cure – treatment include dopamine replacement therapy, L-DOPA, dopamine agonists,
MAO-B inhibitors
● Why is Parkinson’s disease a good candidate for stem cell therapy?
○ Localised degeneration of a neural pathway
○ Ability to generate/harvest large numbers of dopaminergic neurons for
transplantation
○ Readily detectable improvements in motor symptoms such as tremors
○ Demonstrated to work with transplantation of foetal mesencephalic or adrenal grafts
with partial restoration of motor function
● Sources of stem cells for Parkinson’s disease
○ Human foetal neural stem cell – isolated neuroectoderm derived germinal zone
(ventricular zone, hippocampal dentate gyrus) *multipotent are difficult to
differentiate into DA neurons (dopaminergic neurons)
○ Human adult neural stem cell – unlikely to obtain large numbers for
transplantation
○ Induced pluripotent stem cells (iPSCs) – the future of stem cell therapy, can be
grown for large scale production
Planarian flatworms – the ultimate regeneration
● Only a few cm in length and live in freshwater
● Can completely regenerate and grow an entirely new worm from an amputated part
● Process requires a population of proliferating cells called neoblasts (adult pluripotent stem
cells)
● Some neoblasts have the capacity to produce a large number of descendent cells through
the process of cell division clonogenic neoblasts
Potential side effect of using stem cells, could differentiate incorrectly, forming a cancerous
tumour
 Lesson 11 - Neurodevelopment
Development 1
Learning Objectives:
● Identify the developmental origin of the central nervous system (CNS).
● Describe the sequence of events in the development of the central nervous system; namely
neural induction, neurulation, neurogenesis, neuron migration, axon guidance and
synaptogenesis.
● Discuss developmental events that continue postnatally with regard to motor control.
● Identify the developmental origin of glial cells in the central nervous system.

Gastrulation
● The epiblast forms three layers – ectoderm (containing the neural plate), mesoderm
(containing the noto cord), and the endoderm
● Neural tissue develops from the ectoderm, the top layer of the embryo.

Neural induction
● Occurs following gastrulation
● The neural plate border has a bunch of signals. As neurulation is forming, the signals
converge, inducing the cells in the neural plate to become neurons and non-neural
ectoderm cells to form epidermis
 ● Neurulation
○ The neural plate folds, forming the neural tube
■ The neural tube joins up first in the middle, and zippers up to closure points,
and if it fails to close completely, neural tube defects occur
○ The centre (lumen) of the tube is filled with fluid.
○ Formation of the neural tube & neural crest make up the CNS (i.e. neurons of
spinal cord) & PNS, respectively.
■ Occurs when the neural plate in the ectoderm folds up and forms the
neural tube

Neurogenesis
● Progenitor cells proliferate to become neuroblasts.
● Cells divide with the nucleus at the ventricular surface.
● Symmetric division produces two or more progenitor cells
● Asymmetric division produces a progenitor cell and a neuron.
○ Asymmetric neurons ensure we don’t run out of progenitors
○ Asymmetric division type 1 – when two new cells are made, one turns into an
apical progenitor cell, and the other turns into a neuroblast than a neuron
○ Asymmetric division type 2 – same as before, but before turning into a
neuroblast, it forms a basal progenitor cell

● Neural migration
○ Radial migration
 ■ Neuroblasts generated from apical progenitor cells migrate along the
process of the apical progenitor cell to the cortex.
■ This is atype of migration where neurons follow a trajectory that is
perpendicular to the ventricular surface, moving alongside radial glial fibres
and expanding the thickness of the neural tube

○ Cortical layering
■ Neurons migrate to form the cortex in an inside-out manner, where the
newest/youngest neurons are distributed on the outer layer ( matter) of
the cortex, while the oldest neurons are distributed on the inner layer (
matter)
● Neurons in layer I are the youngest (born later) - nearest to cortical
surface (apical)
● Neurons in layer VI are the oldest (born earlier) - nearest to
ventricular surface
■ Neurons (i.e. pyramidal neurons) in layer V are the oldest (born earlier)
compared to neurons in layer II.

Neural patterning
● The biological process by which cells in the developing nervous system acquire distinct
identities according to their spatial positions
● Different parts of the neural tube swell to become ventricles, which form different parts of
the brain (fore/mid/hind), with different signals (growth factors) occurring in different places
leading to the formation of different neural patterns in the brain and spinal cord
○ Progressive patterning over time
○ Results in the patterning of the cortex – different expression patterns of particular
transcription factors in different regions and layers
○ Different expression patterns of particular transcription factors in different
regions/layers controls the patterning of pyramidal neurons which happens
progressively over time

● Axon growth
○ Pyramidal motor neurons need to synapse on motor neurons in the spinal cord.
○ Extend out processes, called neurites
○ Growth cone on developing axon

● Axon guidance
○ Neurons extend a growth cone at the end of the axon
○ Growth cone – express many receptors that bind to environmental ligands, with the
intracellular response to these ligands affecting the behaviour of the growth cone
■ A growth cone is a large, actin-supported extension of the developing or
regenerating neurite seeking its synaptic target and is found on the axon
○ Environmental signals control axon guidance
■ Come from substrate (i.e. extracellular matrix, other cells)
■ It is the guidance cues
➢ Attractive signals → growth: promote the growth of actin filaments,
thus the growth cone will extend that way
➢ Repulsive signals → retraction: depolarises the actin filaments and
causes them to ‘chop off’
➢ Long range → broad reach
➢ Short range → small reach
➢ Severed microtubules → retraction
■ E.g. Adding a chemorepellant near the target site of a group of axons,
during axon guidance, what would likely to happen is many growth cones
would retract and not enter the target site
● Synaptogenesis
○ Neurons make contact
○ Contact stabilisation by cell-cell adhesion
 ○ Synaptic maturation – recruit synaptic machinery to the site of the immature
synapse
○ Neural activity regulates (strengthens/weakens) the synaptic connection via
neurotransmitters. *If the neurons miss their synaptic target, they undergo apoptosis
○ Synapses are plastic, and change over time
■ Activity is critical in maturation of synapses, and whether they are retained
or eliminated.
■ Plasticity and learning.

Later development and postnatal changes

● Gliogenesis
○ Begins after neurogenesis is finished
○ Developmental process by which glial cells – astrocytes, oligodendrocytes (from
neural plate), Schwann cells (from neural crest cells), microglia (from immune
system, later migrate into brain) – are generated.
○ It includes the production of glial progenitor cells and their differentiation into mature
glia.
● Dendritic spine growth
○ Dendritic spines can develop directly from dendritic shafts. During the first week of
birth, the brain is predominated by dendritic protrusions, which eventually develop
into synapses. However, after the first week, the dendritic protrusions are replaced
by spiny dendrites but also, small, stubby spines that protrude from spiny dendrites
○ Development of dendritic spines
■ Pyramidal neurons extend an apical dendrite and multiple basal
■ Length increases after birth (postnatally)
■ Dendrite spines develop over time
■ There are different types of spine shapes
○ Synaptic connections change over time
■ With age, dendrites get prunes and eliminated
■ Number of spines changes with time
■ Important part of development/maturation

● Development of electrophysiological function

○ Electrophysiological properties develop early, but undergo significant changes in
maturation.
● Prenatal brain development
 ● Brain development
○ Important processes in brain development continue after birth.
○ Development of brain can be influenced by genes and environment.

Development 2
Learning Objectives:
● Describe the origin of the peripheral nervous system.
● Describe the events involved in development of the sympathetic ganglia, including the
epithelial-mesenchymal transition and migration of the neural crest.
● Identify the developmental origin of glial cells in the peripheral nervous system.
● Explain advantages and disadvantages of using organoids to study brain development.
Neural crest cells induction/formation
● *Neural crest cells express different genes to the neural plate and neural tube
● Neural plate border cells induced to become neural crest cells and further form neurons
of the glia and peripheral nervous system of many cell types
○ Dorsal root ganglia
○ Sympathetic and parasympathetic ganglia
○ Enteric nervous system
● In order for the neural crest cells to migrate and break free from neural tube, they must
undergo epithelial-mesenchymal transition. *Neural crest cells become mesenchymal
and migrate from the neural tube
○ Epithelial-mesenchymal transition involves changes in:
■ Decreased expression of cell-cell adhesion (less)
■ Decreased polarity along apical-basal axis (less)
■ Retraction of cell shape (changes from round to irregular)
■ Decreased cell-matrix adhesion (less)

Neural crest cells migration

● What do neural crest cells form?
○ After induction, neural crest cells migrate away from the neural tube and follow
numerous pathways to their target sites
○ Neural crest cells form Schwann cells, dorsal root ganglia, sympathetic
ganglia, pigment cells, adrenal medulla (i.e. Neural crest cells in the trunk form)
■ Dorsal root ganglia – perpendicular to the spinal cord
■ Sympathetic ganglia – parallel to the spinal cord
● Sympathetic ganglia chain formation
1. Neural crest cells migrate via somites towards the dorsal root aorta
2. Cells are dispersed along the aorta cells (no segregation or aggregation)
3. Cells aggregate into discrete ganglia (collection of cell bodies)
■ *Increased increased cell-cell adhesion can mediate the aggregation of
cells of the sympathetic ganglia
■ Neurite outgrowth
■ Axon growth and guidance to peripheral targets.
 4. Sympathetic ganglia move to a more dorsal position, towards the spinal cord
5. Preganglionic neurons synapse with ganglion cells and ganglion cells synapse with
targets (creating the chain)
○ Neural crest cells become mesenchymal and migrate from the neural tube - a
critical step in the development of sympathetic ganglion neurons

Neurotransmitter plasticity
● Most sympathetic ganglion neurons use noradrenaline as a neurotransmitter.
● Some use acetylcholine.
● Some neurons can change phenotype (E.g. from adrenergic to cholinergic)

Myelination and glia in the periphery

● Gliogenesis at same time as neurogenesis in the PNS
● Schwann Cells and Satellite Cells develop from the neural crest

Tumours from the peripheral nervous system

● Sympathetic ganglion cells can give rise to neuroblastoma
● Cancer of the sympathetic ganglion cells most commonly occurring the adrenal medulla
○ Neuroblastoma most commonly develops from neuroblasts in adrenal medulla
● It is the most common solid tumour in childhood.

Animal models to study brain development

Drosophila (flies) Discovery of axon guidance molecules

C. elegans (worms) Role of specific neurons and axon regeneration

Zebrafish Adult neurogenesis

● Model for numerous neurodevelopmental disorders

Mouse ● Various genetic tools
● Mammal

Human organoids to study brain development

● Human organoids are human embryonic stem cells that are exposed to growth factors,
which can cause them to develop into an organ
● Features of neural development is currently difficult to study using human brain
organoids: Vascularisation of the brain
Pros Cons
● Might be better able to model human ● Not exact replicas of human organs
organs than animal models ● Lack vascularisation and an immune
● Can be used to study evolution, system (microglia)
specific patient mutations, brain cancer ● Ethical issues
(e.g., glioma)
 Lesson 12 - Pharmacology
Drugs affecting autonomic neurotransmission
Learning Objectives:
● Identify sites where drugs can act to modulate the neurotransmission process & the
mechanisms by which they do this
● Explain the advantages/disadvantages of these sites as therapeutic targets
● Predict the effects of drugs that act at these sites

Autonomic Nervous System (ANS)

● Part of the peripheral efferent nervous system
● Parasympathetic + Sympathetic NS
○ most tissues innervated by both arms of the ANS
○ Each system tends to oppose or balance the other
● Tissues are under involuntary control
Parasympathetic NS Sympathetic NS

Rest and Digest Fight or Flight

● Cell bodies are located in the ● Cell bodies are located in the cranial
thoraco-lumbar region of the spinal sacral region of the spinal cord
cord ● Long pre-ganglionic, short
● Short pre-ganglionic and long post-ganglionic
post-ganglionic ● Both pre and post-ganglionic neurons
● Pre-ganglionic neurons release release Acetylcholine (ACh)
Acetylcholine (ACh)
● Post-ganglionic neurons release
noradrenaline (NA)

Neurotransmitter Transmission
● Contact Dependent - membrane-bound signal molecule
● Paracrine - local mediators (neurotransmier) (nearby cells)
● Synapc - axon innervang a target cell via synapse
● Endocrine - via the bloodstream (hormones)

● Summary of neurotransmission
 1. Start with the uptake of the transmitter entering the nerve terminal via active
transport
2. Precursor is activated by metabolised enzyme to form the neurotransmitter
3. Neurotransmitter is stored within the storage system (vesicle)
4. Action potential arrives at axon terminal – causing influx of calcium and release of
neurotransmitter
5. Neurotransmitter interacts with receptor/effector organs
○ Determination of the action of the transmitter:
■ Inactivation of the transmitter
■ By metabolism
■ Taken back by the neural tissue by active uptake
■ Taken up by non-neuronal tissue

Peripheral nervous systems (PNS)

● Autonomic nervous system (ANS)
○ Innervates smooth muscle/cardiac muscle
○ Regulates activity of various organs (e.g. heart gut pupil, airways, blood vessels etc)
○ NON-voluntary control
■ Two neurons: preganglionic and postganglionic neurons – innervate blood
vessels etc.
■ Parasympathetic
➢ ACh → ACh (salivary glands etc.)
■ Sympathetic
➢ ACh → NA (blood vessels etc)
 ➢ ACh (ONLY ADRENAL MEDULLA)
● Motor nervous system (neuromuscular) (somatic)
○ Innervates skeletal muscle only
○ Regulates neuromuscular activity/coordinates all main muscle groups and function
○ Voluntary control
■ One neuron: 1a motor neuron to skeletal muscle

Adrenal Glands
● Located above each kidney
● Each gland is composed of an inner part (adrenal medulla) and an outer layer (adrenal
cortex)
○ Adrenal medulla → adrenaline (epinephrine)
○ Adrenal gland is activated by the sympathetic system and secretes adrenaline INTO
THE BLOODSTREAM
○ It’s this that can relax and dilate bronchi in parasympathetic response

Important things to remember

● Some tissues are only innervated by one side of the ANS
○ Bronchi in lungs: only innervated by the parasympathetic nervous system but
respond to adrenaline
○ Blood Vessels: only innervated by the sympathetic nervous system
● Sometimes there is more than one transmitter
○ Co-transmitters are stored with, and released with the neurotransmitter. They have
their own receptor and modulate activity
■ Sympathetic NS = ATP and NA
■ Parasympathetic NS = NO and Ach
● In parts of the ANS the main transmitter is neither acetylcholine (Ach) or noradrenaline
(NA) and includes the NANC system (Non Adrenic Non Cholinergic)
○ E.g., Nitric oxide (NO) is involved in erection
● Sweating and sweat glands is tricky
○ Sweat glands are part of the sympathetic nervous system and are cholinergic
meaning that they use the neurotransmitter acetylcholine which is characteristic
of the sympathetic nervous system
Effect of Parasympathetic NS Effect of Sympathetic NS

● Pupil constriction ● Pupil dilation

● Bronchoconstriction ● Contraction of radial muscle
● Decrease in heart rate ● Bronchodilation (no direct intervention,
● No effect on blood vessels adrenaline released can affect the lung)
● Increased GI motility (relaxes ● Increase in heart rate and force
sphincters) contraction
● Stimulates exocrine secretions (tears, ● Vasoconstriction (increase in blood
saliva, etc.) pressure) and vasodilation (to muscles)
● Decreased GI motility (constricts
sphincters)
● Sweating

Bronchi in lungs: only innervated by the Blood Vessels: only innervated by the
parasympathetic nervous system but respond sympathetic nervous system
to adrenaline
Receptors of the ANS
● Receptors for Acetylcholine (ACh) - parasympathetic nervous system
○ Cholinoreceptors (AChr)
■ Nicotinic ACh receptors (nAChRs)
➢ Found on on ganglia and on skeletal muscle

■ Muscarinic ACh receptors (mAChRs)

➢ Found on tissues and responsible for organ effects (e.g. heart)

➢ Effects seen when muscarinic receptors are stimulated

(DUMBBELLS):
- Pupil constriction (Miosis)
- Stimulated saliva flow (Salivation)
- Decreased heart rate (Bradycardia)
- Bronchoconstriction
- Stimulates peristalsis and secretion (Diarrhoea)
- Stimulates bile release
- Bladder constriction (urination)
- Emesis (vomiting due to increased gut activity)
- Lacrimation (increased tear production as lacrimal glands
(tear glands) are stimulated)
- Lethargy (fatigue or drowsiness)
● Receptors for Noradrenaline (NA) - sympathetic nervous system
○ Adrenoreceptors
■ α-adrenoceptors (alpha)
➢ α1, α2
➢ Found on tissues and responsible for organ effects
➢ Excite and contract (in the GIT they close sphincters stop food from
moving through the gut)
■ β-adrenoceptors (beta)
➢ β1, β2
➢ Found on tissues and responsible for organ effects
➢ Eelax and inhibit (except in the heart where they stimulate)
■ Location of adrenoreceptors
➢ Sympathetic nervous system
- Adrenergic
- Involuntary
➢ Smooth muscle and cardiac muscle

■ What effects are seen when peripheral adrenoreceptors are

stimulated?
➢ Mydriasis (pupil dilation) and reduced saliva flow (alpha)
➢ Increased stroke volume and heart rate (beta)
➢ Vasoconstriction (alpha)
➢ Reduced peristalsis and secretion (alpha and beta)
➢ Glycogen converted to glucose (beta)
➢ Inhibition of bladder contraction (beta)
➢ Bronchodilation (beta, not innervated)
➢ Adrenaline release
 α-adrenoceptors (alpha) β-adrenoceptors (beta)

Noradrenaline > adrenaline > isoprenaline Isoprenaline > adrenaline > noradrenaline
● Excitatory ● Inhibitory
● More involved in contraction ● Relaxation / dilation
● Found in smooth muscle only ● Exception = the heart
● Found on the heart, lungs
(bronchodilation), and vascular smooth
muscle tissue

● *Both systems can have some effects, however, are mostly affected by the dominant
system
○ Tissues may have receptors for both Ach and NA receptors may be present even if
nerves aren’t
○ E.g., NA receptors in bronchial smooth muscle and ACHR receptors in vascular
smooth muscle “autonomic receptors” may exist in other parts of the body not
associated with the ANS including the brain

Drugs can act at various stages of neurotransmission:

1. Synthesis
2. Storage
3. Release
4. Action at receptors
5. Termination of action
● Summary of neurotransmission
1. Start with the uptake of the transmitter entering the nerve terminal via active
transport. Precursor is activated by metabolised enzyme to form the
neurotransmitter
2. Neurotransmitter is stored within the storage system (i.e. in synaptic vesicles)
3. Action potential arrives at axon terminal – causing influx of calcium and release of
neurotransmitter. The release of transmitter through the process of Exocytosis.
4. Neurotransmitter interacts with receptor/effector organs
5. Action of the transmitter at the receptor must be terminated either by being broken
down by enzymes found within the synapse or reuptake by being removed from
the synapse by being sucked up & taken up & recycled back into the nerve terminal.
■ Dermination of the action of the transmitter:
➢ Inactivation of the transmitter
➢ By metabolism
➢ Taken back by the neural tissue by active uptake
 ➢ Taken up by non-neuronal tissue

Targets for drugs

● Drugs tend to work because they’re able to bind to protein targets and may mimic or inhibit
the actions of endogenous factors such as transmitters.
● A drug will not work unless it is bound.
○ Most* drugs exert their effects by binding to protein molecules
■ Enzymes
➢ Cyclo-oxygenase (COX1 and COX2) - NSAIDs
■ Transporters
➢ SERT (5HT transporter) SSRIs
■ Receptors
■ Ion channels
➢ Na+ channels - local anaesthetics
○ Agents/drugs binding to protein targets can
■ Mimic the actions of endogenous factors (e.g. transmitters)
➢ Most common at receptors targets where drugs mimic the actions of
the neurotransmitter
■ Inhibit the actions of endogenous factors
➢ Many drugs are enzyme inhibitors or transport blockers
➢ Many drugs can also BLOCK receptors preventing the action of
neurotransmitters
○ Agents/drugs targeting receptors
■ Agonists - MIMETICS
➢ Agents that mimic the actions of the transmitter are called agonists
- ParasympathoMIMETICS - Agonists that prefer acting on
parasympathetic system
- SympathoMIMETICS - Agonists that prefer acting on
sympathetic system
■ Antagonists – LYTICS
➢ Agents that prevent the action of neurotransmitters are called
antagonists
- ParasympathoLYTICS - Antagonist that prefer removing the
effect on parasympathetic arm of ANS
- SympathoLYTICS - Antagonist that prefer removing the effect
on sympathetic arm of ANS

Drugs affecting parasympathetic system

● Targeting Ach receptors
○ WHAT EFFECT WILL MUSCARINIC AGONISTS CAUSE?
■ Pupil constriction
■ Stimulated saliva flow
■ Bronchoconstriction
■ Decrease in heart rate
■ Increase gastrointestinal (peristalsis and excretion) → Increased GI motility
= diarrhoea
■ Bladder contraction
■ Bile release
○ WHAT EFFECT WILL MUSCARINIC ANTAGONISTS CAUSE?
■ Pupil dilation
■ Inhibit secretions (dry mouth)
■ Increased heart rate
■ Relax bronchial smooth muscle (bronchodilation)
■ Decreased gastric motility and gastric acid secretions → slowing of GI
movement = constipation
■ Decreased bladder emptying
○ CLINICAL USES OF DRUGS TARGETTING THE PARASYMPATHTIC SYSTEM
■ Drugs targeting muscarinic receptors – eye
➢ Agonists:
- Cause pupil constriction and decrease intraocular pressure
(used on glaucoma which is a condition with increased
pressure in the eye)
➢ Antagonists:
- Cause mydriasis/pupil dilation (e.g., eye examination)
■ Drugs targeting muscarinic receptors – lungs
➢ Asthma and COPD
➢ Agonist:
- Causes bronchoconstriction (no clinical need)
➢ Antagonist:
- Causes bronchodilation/decrease bronchial smooth muscle
(e.g., chronic obstructive pulmonary disease)
- Dry up mucous
➢ Anaesthetic premedication (dry secretions):
- Muscarinic agonist – associated with exocrine secretion
primarily stimulated in the respiratory tract
- Muscarinic antagonist – inhibits secretion which can be useful
in surgery
■ Drugs targeting muscarinic receptors – GIT
➢ Agonists:
- Increase motility (may be useful to treat constipation)
➢ Antagonists:
- Decrease motility (used to treat diarrhoea)
● Drugs affecting Ach neurotransmission
○ Synthesis and storage of acetylcholine (ACh)
1. Choline comes from the diet and enters the axon terminal by active transport
through choline transporter
2. Acetylcholine (Ach) is synthesized in nerve terminals from choline by the
enzyme choline acetyltransferase (ChaT).
 ➢ Choline acetyltransferase (ChaT) synthesizes acetylcholine
➢ Acetyl-CoA + choline —ChaT→ CoA + ACh
3. Ach is stored in vesicles & released via exocytosis into the synaptic cleft
where it can bind to muscarinic receptors and elicit a response
4. AP arrives at axon terminal resulting the influx of calcium and fusion of the
vesicle to the terminal membrane followed by the release of Ach
5. Action of ACh at receptors terminated by the enzyme acetylcholinesterase
(AChE), which breaks it down into its constituent parts, one of which is
choline that can be reused for new synthesis of acetylcholine.
➢ Acetylcholinesterase (AChE) degrades acetylcholine.
➢ ACh —AChE→ acetate + choline
○ What would be the effects of an agents that prevents ACh breakdown?
■ Drugs that inhibit acetylcholinesterase, known as anti-cholinesterases,
prevent the breakdown of acetylcagentholine.
➢ This leads to an increase in acetylcholine at the synaptic cleft,
enhancing its effects.
■ Inhibiting acetylcholinesterase activates the parasympathetic system can
lead to over-activation of the parasympathetic system, causing bigger
DUMBBELLS effect
➢ Miosis (pupil constriction)
➢ Increased salivation
➢ Slowed heart rate
➢ Bronchoconstriction
➢ Increased gastrointestinal motility
○ Inhibitors of ACh breakdown are found in organophasphate insecticides what
would you expect in an individual?
■ Organophosphates, found in some insecticides, are a type of
anti-cholinesterase (i.e. inhibit cholinesterase enzymes & thereby
preventing the breakdown of acetylcholine)
■ Organophosphates poisoning can
➢ Excessively activate parasympathetic system
- DUMBBELLS effect
➢ Influence skeletal muscle & neuromuscular system (i.e. somatic
nervous system)
- Acetylcholine is a neurotransmitte at neuromuscular junctions
- Muscle overactivity and subsequent paralysis.
○ Therapeutic use of anti-cholinesterases
■ Myasthenia gravis (NMJ) – autoimmune condition associated with
decreased skeletal neuromuscular transmission of Ach
➢ Muscle weakness due to insufficient effects of ACh at skeletal
muscle
➢ Little bit of anti-cholinesterases can help strengthen neuromuscular
transmission by preventing the breakdown of ACh.
■ The use of anticholinesterases results in unwanted side effects
(DUMBBELLS) and overtreatment may lead to cholinergic crisis
○ What would be the effects of an agents that interferes with ACh release?
■ BOTOX – botulinum toxin
➢ Cause of some form of food poisoning and inhibits the release of
Ach by interfering with exocytosis as it prevents the fusion of
vesicle with the membrane
 ➢ Anti-DUMBBELLS effect:
- Mydriasis (pupil dilation)
- Dry mouth
- Increased heart rate
- Bronchodilation
- Slowing of GI movement = constipation
➢ Affecting neuromuscular system, which also uses acetylcholine
- Progressive muscle weakness & paralysis
- Drooping eyelids
- Slurred speech
- Difficulty breathing that may lead to respiratory failure as
diaphragm is affected
- Facial weakness on both sides of the face
■ If botulinum toxin affects so many sites, can it have a therapeutic use?
➢ Injected directly to the target tissue
➢ Selectivity of action potential to ensure it’s not getting into the
circulation
■ Therapeutic uses of Botox
➢ Mostly related to effects on skeletal muscle neurotransmission
- Botox does not have a use in any therapeutic benefit in
targeting the parasympathetic system.
➢ Bleopharospasm – uncontrolled contractions of the eyelids (relaxes
the muscles to be kept open)
➢ Cerebral palsy – not an approved use (reduces muscle rigidity and
uncontrolled spasms)
➢ Hyperhidrosis – excessive sweating (injected directly into sweat
gland to cause reducing in ACh)

Drugs affecting sympathetic system

● Targeting NA receptors
○ WHAT EFFECT WILL NA AGONISTS CAUSE?
■ Simulation of α-adrenoceptors receptors
➢ Vasoconstriction – raising blood pressure
➢ Slowing gut
➢ Mydriasis
■ Simulation of β-adrenoceptors receptors
➢ Increasing force and rate of heart contraction
➢ Slowing gut
➢ Relaxing and dilation of bronchial smooth muscle
○ WHAT EFFECT WILL NA ANTAGONISTS CAUSE?
■ BLOCKING α-adrenoceptors receptors
➢ Vasodilatation
➢ Lowering blood pressure
➢ Speeding gut
➢ Miosis
■ BLOCKING β-adrenoceptors receptors
➢ Decreasing force and rate of heart contraction
➢ Speeding gut
➢ Constriction of bronchial smooth muscle
○ CLINICAL USES OF DRUGS TARGETTING THE SYMPATHTIC SYSTEM
 ■ Drugs targeting α-adrenoceptors – eyes
➢ Agonists:
- Cause pupil dilation
- Useful in eye examination and surgery
- There’s also a related effect on blood vessels – drugs that
activate alpha receptors also cause vasoconstriction and
produce decreased reddening of the eye
■ Drugs targeting α-adrenoceptors – blood vessels
➢ Agonists:
- Cause vasoconstriction
- Constrict small blood vessels in nose and eye acting as nasal
decongestants and ocular decongestants
- Given “locally” only
➢ Antagonists:
- Relax blood vessels in circulatory system
- Used in hypertension
- Given so to access all these blood vessels
■ Drugs targeting β-adrenoceptors – heart
➢ Agonists:
- Stimulate the heart
- Can be used in heart failure
➢ Antagonists:
- Slow the heart
- Can be used to help lower blood pressure indirectly
➢ “beta blockers”
■ Drugs targeting β-adrenoceptors – bronchi/lungs
➢ Agonists:
- Relax and open bronchi
- Used in asthma
➢ Antagonists:
- Contraindicated in asthma
- Would trigger asthma
● Drugs affecting NA neurotransmission
○ Process of transmission of noradrenaline (NA) at sympathetic nerve terminals
1. L-tyrosine (precursor) from diet
2. Metabolised to dopamine
3. In the vesicle, the final stage of NA synthesis occurs
4. Vesicles contain: noradrenaline (NA), dopamine (DA) and ATP
5. Action potential and calcium influx
6. Noradrenaline acts on alpha or beta receptors depending on the location
7. The action of noradrenaline and receptors is terminated by REUPTAKE
➢ Reuptake to the axon terminal (primary)
- NA actions are removed from the receptor by the nerve
terminals, sucking it up back up into the nerve terminal
through this pump called uptake-1
➢ Reuptake by non-neuronal tissue (secondary)
○ What would be the effects of an agents that prevents NA RE-UPTAKE?
■ NA re-uptake blockers are used in depressive disorders (effects are in the
brain rather than the ANS)
■ Noradrenaline reuptake inhibitors (NRIs) are a type of drugs that work by
increasing noradrenaline amount in the brain.
➢ Works by blocking the reuptake of NA, which is a process that
removes neurotransmitters (i.e. NA) from the synapse (the gap
between nerve cells) after they have sent their signal.

➢ In the PNS, or sympathetic nervous system, blocking the reuptake

of noradrenaline can lead to an increase in sympathetic
transmission.
- Increased HR
- Increased BP
➢ In the CNS, or the brain, NRIs can lead to effects such as excitation
and euphoria
- Because NA plays a role in mood regulation, and increasing
its levels can help alleviate symptoms of depression
■ Actions of cocaine and amphetamine at dopamine (DA) synapses in
the brain
➢ Cocaine works by blocking the reuptake of DA neurotransmitter,
leading to increased DA levels and a heightened response.
- An uptake blocker preventing the removal of DA
➢ Amphetamines cause a large release of DA into the synapse, also
leading to increased DA levels and a heightened response.
- Release of DA
- Indirectly acting mimetic
 ○ Drugs affecting NA release
Direct blockers Releaser

● Noradrenergic neuron blocking drugs ● Includes amphetamine,

○ Abolish response to nerve stimulation ephedrine, pseudoephedrine
○ Interfere sympathetic NS response and (structurally related to NA
reduces reflexes weak direct action of NA
● Mechanism of action? receptors)
○ Not well understood ● Mechanisms of action?
○ Requires the uptake of the drugs into the ○ Indirectly acting
nerve terminal sympathomimetics
■ Happens by same neuronal ○ NA released
uptake NA uses independently of AP
○ Once inside the nerve it interferes with ○ Activation by indirect
the movement of AP in the nerve terminal release of NA
and depletes the NA from vesicles
meaning there is less NA released to
interact with receptors
● Effect NA release from all sites of the body
● Bretylium guanethidine
○ Noradrenergic neuron blocking drugs
used for treatment of hypertension.
Bretylium inhibits norepinephrine release
by depressing adrenergic nerve terminal
excitability

○ What would be the effects of agents that act as an indirectly acting mimetic at
the sympathetic system?
■ Sympathomimetics
➢ Substances that mimic or modify the actions of endogenous
catecholamines of the sympathetic nervous system
➢ Modes of transmission:
1. Amphetamine ephedrine enter nerve terminal uptake
2. Once inside they are taken up into vesicles by exchange with
NA
■ Pseudoephedrine
➢ An indirectly acting sympathomimetic drug, which means it
mimics the effects of the sympathetic nervous system
- Indirectly acting mimetic
➢ Cause vasoconstriction of dilated nasal vessels by activating
α-adrenoceptors
➢ Leads to decreased swelling and nasal congestion
- Nasal decongestant
■ Amphetamine and related compounds
➢ ADD (ADHD)/ recreational use (dopamine effects)
 ➢ Side-effects: tachycardia, hypertension

Drugs affecting ganglionic neurotransmission

● Targeting Nicotinic (nACHRs) receptors
○ nACHRs are found
■ In ganglia on the adrenal medulla
■ On neuromuscular receptors (i.e. Found on skeletal muscle and are
associated with the contraction of muscle)

○ Activation or blockade of nACHRs

■ Will affect ganglionic neurotransmission
■ Responses complicated by affecting both the sympathetic and
parasympathetic arms