Joseph Ekholm

9/7/2020
Zoology 500
9.2 Nacewicz Essay

From this lecture, I learned, applied, and noted several interesting findings. First, the fact that

Autism is characterized by a social information overload rather than no information was quite

interesting. Additionally, the overgrowth of the amygdala until, on average, 12.5 years old

followed by identical shrinkage is present in individuals with autism, low SES, history of abuse,

and lifelong environmental stress. This finding emphasizes the negative impact that biological

and environmental stress can have on the brain, particularly the amygdala. I am curious as to

how my amygdala may be impacted by my anxious temperament and tendency to be

overstressed. Brendon also noted the typical measurements of glutamate, GABA, inositol, and

glutamine in response to a stressor. The typical glutamate spike was actually an inverse curve in

those with Autism potentially due to the conversion of glutamate to glutamine in the

astrocytes. This finding sparked curiosity in knowing what my own neurotransmitter response

would look like due to my overly stressed and anxious tendencies. Would I have the normal

spike in glutamate or a more inverse reaction as seen in the autism trials? Could observing this

response also help place one on the spectrum of Autism? He also mentioned they use the

neural network to diagnose autism, but this is a curiosity. The last tidbit of intriguing

information was the mention that the sinus passageways were an interference in his

microscopy imaging and that they play a role in buffering of sensory input (ie so you won’t hear

your eyes move in your head which is crazy!).

 Flooding of autonomic nervous system until no longer response
 Autism- didn’t activate amygdala (no understanding? NO overloaded rather than no
information
o Eye tracking in fMRI task
o Fight or flight system
o Eyes overactive
 Karger amygdala more eye fixation
 Smaller  not good and slower judgement
 Stress makes amygdala bigger then metabolic toxicity so shrinks
 Overgrowth first decade then shrinkage
 Identical shrinkage at 12.5 years old on average comparing abuse, low SES and post
inlife long stress
 Size environmental and biological stress
 HOW TO STOP
o Measure glutamate GABA and
o Glutamate high sensitivity for stress response
o Autism model upside down glutamate response
 Astrocytes convert glutamate to glutamine
 Block alpha 1 receptor (PTSD drug that works)
o Spike inositol (adrenaline response)
 Excessive response in autism
 SINUS passages buffer sensory input
 Anxious temperament
o Injection glutamate to GABA increase in gaba and cortisol and metabolism
NOW GOOD MEASURE FOR GENE THERAPY
  Expansion therapy 4000x faster
o Explore social behavior in fear
 Protein kinase C
 Also cells that get oxytocin (social bonding)
 Association between social behavior and fear
 MOM as buffer for social fear
 IP3R3
 PKCd cells get learned fear from amygdala and oxytocin form social behavior same
cells that have IP3R3 to have empathy
Joseph Ekholm
9/14/2020
Zoology 500
9.9 Gammie Essay

From Colleen McDowell’s talk, I learned an extensive amount about Glaucoma, its

pathogenesis, and potential treatments. Also, the scientific method of investigation that

Colleen utilized to progress through her research and discovery was very interesting and noted

important elements and steps in the process of scientific research. Using physiological

understanding, investigation of intraocular pressure as a risk factor and causative factor of

glaucoma led to examination of a exfoliation syndrome model upon recognition of another

researcher. This examination then perpetuated finding of a model system of Ad5 transgene and

an inducible mutant myocilin model to aid in further research. She then moved forward in her

research to investigate mechanisms and signaling pathways. The role of TGFß2 was then

coupled with her previous research to investigate a further modeling and mouse strain

investigation leading to comprehension of the TLR-4 pathway including BAMBI to understand

Glaucoma and optic nerve head damage. This procedural approach then potentiates therapy

targets. Hearing about her research process and how she combined physiological understanding

with complementary research and mouse modeling has educated me on how I can approach

research and scientific investigation. I can apply this processive questioning into any field I want

to research currently and in the future.

NOTES:
1. Glaucoma models
o Irreversible peripheral progressive very widespread
o Optic nerve head cupping
 Axons die and cup and cause loss
  Ganglion loss associated with intraocular pressure
 Trabecular meshwork
o Risk factors
 INTRAOCULAR PRESSURE
 Ocular hypertension (inc aq humor outflow impeded)
 Lower pressure prevents glaucoma
o Common forms
o Screen for new mutants
 Iris phenotypetransillumination
 Exfoliation syndrome model!!!!
 3 BP deletion
o INDUCABLE model
o Homogenous response
o Ad5 transgene expression in Trabecular meshwork for drainage (GFP)
o Inducible mutant myocilin model
o Mechanisms glaucoma (cornea and irisTM resistance but too much
resistanceglaucoma)
2. Discovery of TGFB2-TLR4 signaling in ocular hypertension
o INC in glaucoma in Aq Humor and optic nerve head
o TGFB2 levels IOP inc
o TGFB2 injection to find most susceptible strain
o C3H/HeJ  NO response
 Polymorphism in strain
 TLR4 wildtype normal response so MUTANT is problem
 Injury inflammation tLR4 activation Progressive fibrosis (INC IOP)
o BAMBI downregulation TGFB activity unregulated!
o FEEDFORWARD LOOP for inc TGFB
o IMPORTANT pathways found
o INC TLR4 in human glaucoma patients
o EX VIVO IF you block TLR4 TGFB2 is blocked
o
3. Identification of novel therapy targets in TM
o
4. RGC subtype specific cell death in mouse models of glaucoma

Joseph Ekholm
9/16/2020
Zoology 500
9.16 Luis Populin Essay

The lectures given by Luis Populin were enlightening to me on a topic of interest that I hadn’t

explored or researched. As a student from a family of ADHD prevalence, I have lots of

familiarity with the treatment and the rising frequency of prescriptions. The most fascinating

detail he noted from his research with Macaque monkeys as well as previous human research

was the effect of Methylphenidate on premature responses versus working memory. The fact

that the dose dictates whether memory is optimized at the expense of attention enhancement

or if memory is hindered while attention is enhanced is thought-provoking and lends to the

need for dosage individualization for each patient. Additionally, the link between impulsivity

and the DAT gene and its methylation provided some insight into the inheritance and genotypic

identification of ADHD as well as elucidating the critical role of dopamine transporters in the

disorder and phenotypic expression. This seems to me as a critical finding especially as

someone with a familial genetic predisposition to the disorder. Luis further expanded upon the

functional connectivity of the dopaminergic network in the brain via PET, fMRI, and

electrophysiological analysis. All in all, this presentation revealed an interesting field of research

about a very common disorder and treatment. I am curious whether this methylation of the

DAT gene has increased in prevalence as we see an ever-increasing prevalence of the disorder.

 Macaque homologous prefrontal cortex

 Working memory, associations, conjunctions, and rules; reward signals and
representation of rewards; bias and top-down control; keeping to a task and switching
between tasks (flexibility); attention
 Deficits to brain and analyze brain
o Phineas gage
 Iron change in temperament
 Frontal cortex
o Receives input from many places
o Memory
o NONE from periphery so all altered and modified/processed
 Role hypothesized to allow implement rules
o Don’t want to show interest and don’t want to look
o Invoke ability to take over responsibility and
o Reign in response
 Delay response test
o Two wells-one with food then block with board and monkey remember food
o REMOVE area 46 NO delay task completion
 ADHD
o Catecholemine levels DA and NE
o Adderall and Ritalin
 MILLIONS of prescriptions yet still not understanding on effects on
cognition and physiology
 Rhesus monkey for testing ADHD and highly successful
o Perform well in wild
 Select model due to very comparable prefrontal cortex
 Dopaminergic innervation of cerebral cortex
 Sound proof room with ocular motor task
 Methylphenidate given
 Operant conditioning
 Flash target in four different directions of eye movements
 Fixation spot
 Flash spot then see correct response
 Methylphenidate 3 mg/kgNO PREMATURE rESPONSES
o Not better past 6 mg/kg
 2.5x increase in focus per time

 At point of hinder performance for eye movement response more sustained attention
o TRADEOFF
 Appropriate amount for both (attention but not best performance)
 Water consumption ad lib
o Not result of thirst
 Dose and child seatingIMPROVEMENT
 After teacher reported, ability to remember was assessed in the lab
 FOUND SAME THING
o Memory peaked at lower dose compared to attention peak at higher
 DIFFERENT EFFECT ON dosages
 LONG TERM Study of translational project (The hartwell foundation)
o Get benefits while limiting the side effects
 Study impulsivity many errors of fixation (tiny eye movement)
 Could not link SNP with behavior classification
 ALL downstream of cytosine
o Methylation of sites was measured
o SNP1 CG4 2 subjects had no methylation at all
o CALM had NO methyl groups
o Impulsive had some methyl groups
 Question: Ability of dopamine transporter and receptors in brain
 PET to see dopamine transporter
 FECNT PET
o Highest in certain areas (NONE IN CEREBRAL CORTEX)
o Greater availability for los palatis
o MP affects transmission (basal ganglia especially)
 More noisy and responsive to positive
 Substantia nigra and ventral  only places with dopamine
o Block dopamine transmission
 Functional connectivity resting conditions
o Different doses and fMRIsystematic changes
o Functional connectivity changes
 Area 46 connected
 Prenucles are wth head of caudate
 Precuneus and hippocampus affected by head of caudate manipulation
Joseph Ekholm
9/16/2020
Zoology 500
9.23 Corinna Burger Essay

Corinna’s talk gave a great overview and key insight into how rats can be used for brain

research particularly learning and memory formation. One aspect that was intriguing was the

ability to isolate a single gene, Homer1C, from the 50+ genes differentially expressed by

impaired learners. Then from isolation of this one gene was able to show how knocking out this

one gene could be a causal factor in whether or not the rat had learning impairments. Clearly

this is a remarkable finding; however, I wonder if any of the other 50 genes she mentioned

would result in identical findings. Further, would similar genes be differentially expressed in a

human brain? Could gene therapy be used to induce similar recovery of function in those with

memory formation and/or learning impairments? Clearly these findings in rats give rise to much

understanding of the hippocampus and the action behind long term potentiation, but is this

translatable to humans in any extent? I believe there is importance of environmental

enrichment in the potentiation of cognitive function in younger humans and throughout life to

maintain this cognitive capacity, so could Homer1C, if critical in humans, also aid in

maintenance of memory formation and learning throughout life? I look forward to following

this line of research and seeing where it can play a role in human modeling and augmentation.

 Learning and memory formation in aged rats

 Find genes that are differentially expressed
 o Superior learners vs. impairment
 HOMER1=protein in postsynaptic density
 Glut1 metabotropic receptor
 Down reg in impaired mice
 Lack Homer1c gene everywhere
o Homer1c sufficient for hippocampus function?
 Adeno-associated virus
o Complete developmental control
o Complete anatomical specificity
o rAAV capable of long-term transduction in tissue
o Unilaterally other hemisphere
o Produced in one week
o TRANSGENIC RATS
o Gene therapy no known pathology
 Stereotactic injection sites for dorsal hippo
 Inject viral vector to coordinates
  transcribe protein expression
 Inject Homer 1c transgene in hippocampal phenotype
 Green fluorescent protein to control for surgery and transgene expression (specific)
 Water maze
o 8 arms
o Visible
 REMOVE VISIBLE GFP stays the same
 HOMER injected  learning curve
 Improve long term potentiation (LTP)
 Stim to see baseline action potential waves when learning
o Change in slope of action potential=excitatory postsynaptic potential
o Stays potentiated=long term potentiation
 Specific paradigm
o mGluR dependent LTP
o Short term to long term
 Agonist rescued
 HOMER1c KO have deficits that can be rescued via gene delivery of Homer1c to
hippocampus
 Super age learners vs aged learners
 Showed approach could work DIFF expression in 50 genes
 FIRST SEGREGATE to impaired vs non-impaired via memory task
o INJECT HOMER1c
o Object location memory task
o Explore 5 minutes
o Hippocampal development
 Morris water maze drop in maze explore to find platform ifnot place on platform
 o Multiple times to find platforms
 Before and after treatment
o Young
o GFP
o H1C
o AGE impaired about 50%  SIGNIFICANT expression
 Summary segregate on OLM task
o Gene delivery of Homer1c to hippocampus of aged impaired animals sig improve
performance on OLM; marginal in MWM

PART 2
 Age related around 12-24 months
 Superior vs impaired learner groups
 Young great variability
 SUMMARY
o 3 month old separated into inf and sup via MWM
o Repeat after 6 weeks w/o memory carryover
o DIFF in learning ability are maintained through middle age
o Train from young age can prevent age-related cognitive decline
 Environ enrich
o Activate mGluR 1/5Homer1c dendritic t4rnslation and translation initiation
of important genes
Joseph Ekholm
9/30/2020
Zoology 500
9.30 Cynthia Kelm-Nelson Burger Essay

The lecture from Cynthia Kelm-Nelson was extremely fascinating and gave great insight into the

use of rat modeling of degenerative disease, specifically Parkinson disease. I found it truly

ingenious that typical symptoms and early markers like vocal impairments seen in humans with

Parkinson’s were able to be translated into the mouse model. For example, vocal deficits found

in humans which can be assessed fairly easily in humans could be evaluated in a different

manner for rats, such as analysis of the larynx as well as Ultrasonic vocalization and

socialization. This results in a complex and innovative tool to study the disease in vitro without

harm done to human Parkinson’s patients. Her findings continue to potentiate a higher

understanding of the disease and how we might diagnose, monitor symptoms, and treat the

disease in the future. I am interested to what extent we can generalize all rat findings to

humans, such as in sex differences or temporal progression of the disease and onset of deficits.

Further, I am interested in similar research in the field to monitor other degenerative diseases

such as Alzheimer’s, dementia, macular degeneration, etc. I know my lab uses rats to model

degenerative neural retina diseases, so I am sure the rat model is widely utilized in the scientific

community. I look forward to following her research and findings in the field to ne day help

humans suffering from Parkinson disease.

 Modulation of complex emotions

 Vocal communications
 Parkinson’s=degenerative
o Whole body
 Substantia nigradopamine
 Protein aggregation
 Basal ganglia
 Voice deficits
o Sig clinical issue
 Signs well before diagnosis
 80-90% DA neurons loss before
 Vocalizations first signs
 Pathology before
 USE genetic rat models
 Rat vocalizations
o Freq modulated ultrasonic vocalizations (USV)
o Larynx; vocal folds
o Thyroarytenoid muscle
o Neural control
 Categorize calls
 SHOW at 8 months=robust deficits
 Basa;l ganglia not affected
 Midbrain norepi dysregulated
 Aggregations of protein