Outline & Study Questions: Mitochondrial Diseases

I. The Mitochondrial Genome

A. Properties of mitochondrial DNA (mtDNA)
1. Maternal inheritance
a. ~All mitochondria in the fertilized egg come from the mother.
b. Paternal mitochondria in the sperm are destroyed soon after fertilization.
2. Genes coded for by mtDNA.
a. 13 genes for subunits of electron-transfer chain (ETC) complexes
i. 7 subunits of complex I
ii. 1 subunit of complex III
iii. 3 subunits of complex IV
iv. 2 subunits of complex V (F0F1-ATPase)
b. 22 genes for mitochondrial tRNA
c. 2 genes for ribosomal RNA
i. Genome contains all components necessary for synthesis of protein
genes on mtDNA.
d. Mito are thought to have evolved from symbiotic bacteria
i. Mito ribosomes are similar to bacterial ribosomes.
ii. Mito protein synthesis is inhibited by antibiotics that target bacterial
ribosomes (tetracyclin, streptomycin, erythromycin, &
chloramphenicol).
3. Other distinctive properties of mtDNA.
a. Higher mutation rate than nuclear DNA.
i. Decreased ability for DNA repair in the mitochondria.
ii. High levels of ROS production in mitochondria leads to somatic
mutations.
b. Each mitochondrion contains multiple copies of the 16 kB mitochondrial
genome.
i. Each cell also contains many hundred mitochondria.
v. Each cell therefore contains many thousand copies of mtDNA genome.
4. Heteroplasmy
a. Mitochondria segregate randomly during cell division.
i. If fertilized egg contains a mixture of normal and mutant mtDNA, some
daughter cells will have higher mutational load than others.
ii. Can lead to mosaicism if cell with high mutational load is the
progenitor cell for an organ or type of tissue.
b. Mutations may also be somatic, often generated by high ROS production
or inefficient removal of ROS.
i. Deletion & point mutations in mtDNA increase with age.
ii. Oxidized guanosine is higher in mtDNA than nuclear DNA.
II. Manifestations of mitochondrial disease.
A. Genes on mtDNA are only involved in construction of the ETC.
1. Mutations in mtDNA can have detrimental effect only on ETC function.
2. Typically manifested by decreased capacity at high [ADP] (threshold effect)

a. Insufficient flux through the ETC during high energy demand.

i. Leads to accumulation of pyruvate, lactate, fatty acids, & triglycerides.
b. Decreased rate of ATP synthesis.
i. Leads to muscle weakness and exercise intolerance.
c. Increased proliferation of mitochondria to compensate for low ETC &
ATPase activity.
i. Leads to formation of ragged red fibers in muscle cells.
ii. May lead to increased replication of mutant mtDNA, relative to normal
mtDNA (by
unknown mechanism).
iii. This may contribute to the progressive nature of many mitochondrial
diseases.
B. Threshold effect in mitochondrial disease.
1. Clinical expression of disease determined by ratio of mutant to normal
mtDNA in a particular tissue type AND how dependent that tissue is on Ox
Phos.
a. Mutation in mtDNA can only impair function of the ETC.
b. Most cells have ample reserve capacity for energy production by Ox Phos
at high [ADP] and can withstand significant damage to ETC.
c. Energy production will not be impaired until mutant mtDNA reaches a
threshold of 50-90% of total mtDNA.
d. Range of 50-90% reflects how harmful the mutation is and how dependent
the cell is on Ox Phos.
2. Sensitivity of tissue type to mutations in mtDNA.
a. Reflects energy demand and dependence on Ox Phos.
b. Central nervous system is most sensitive.
i. Followed by heart & skeletal muscle > renal > endocrine > liver.
3. Phenotype is determined by how much mutant mtDNA is present (threshold),
which tissue is affected (heteroplasmy & mosaicism) and nature of the
mutation.
a. Defects in muscle cause lactic acidosis, muscle weakness, & ragged red
fibers.
b. Defects in brain & neurons cause myoclonic seizures, ataxia, & stroke-like
episodes.
c. Some patients have combinations of these symptoms.
III. Most common mitochondrial diseases.
A. Lebers Hereditary Optic Neuropathy (LHON).
1. Caused by one of three different mutations in complex I
2. Occurs primarily in young men (not known why there is gender bias).
3. ROS production by mutant complex I is thought to contribute to progressive
nature of disease.
B. Myoclonus Epilepsy with Ragged Red Fibers (MERRF).
1. Caused by mutation in tRNA for lysine.
a. Muscles have ragged red fibers & low activity of complex IV.
b. Not known why mutated tRNALys would preferentially affect complex IV.

2. Symptoms include myoclonus, seizures, cerebellar ataxia due to mutant

mtDNA in neurons, and ragged red fibers & myopathy due to mutant
mtDNA in muscle cells
3. Onset can be childhood (high inherited mutational load) or early adulthood
(low inherited mutational load).
C. Mitochondrial Encephalomyopathy, Lactic Acidosis and Stroke-like Episodes
(MELAS).
1. Caused by mutation in tRNA for leucine.
a. Muscles have ragged red fibers but normal complex IV activity.
b. Not known why the phenotype is different from MERRF.
2. Symptoms also include recurrent headaches, vomiting, dementia, lactic
acidosis and stroke-like episodes.
3. Onset is usually before age 40.
D. Parkinsons disease.
1. Caused by selective destruction of dopaminergic neurons of substantia nigra.
a. Symptoms usually appear in 6th or 7th decade of life and progress slowly.
b. On autopsy, surviving neurons contain protein aggregates (Lewy bodies).
c. Lewy bodies always contain -synuclein, but may contain many other
proteins.
i. Lewy bodies may be protective: attempt by cell to sequester damaged
proteins.
2. Mutation of mtDNA not involved, but impairment of complex I may
contribute.
3. Other contributing factors include mutations in -synuclein, mutations in
proteasome, oxidative stress, and environmental toxins (especially
complex I inhibitors).
4. Inhibitors of complex I can initiate Parkinson-like disease.
a. MPTP contaminant in ecstasy induced Parkinson-like symptoms in young
adults.
i. MPTP converted to MPP+ in astrocytes.
ii. MPP+ selectively taken up by dopamine carrier in dopaminergic
neurons.
iii. MPP+ inhibits complex I and induces oxidative stress.
b. Rats treated with rotenone (complex I inhibitor) develop Parkinson-like
disease.
i. Symptoms include loss of complex I activity, formation of Lewy bodies,
destruction of
dopaminergic neurons.
ii. Thought that impaired complex I causes oxidative stress, which
eventually leads to
apoptosis of dopaminergic neurons.
5. Current hypothesis is that Parkinsons disease may not have a single
underlying cause; several different defects could cause destruction of
dopaminergic neurons by different mechanisms.

Study Questions: Mitochondrial Diseases.

1. Which parent(s) is mitochondrial DNA (mtDNA) inherited from?
2. All of the proteins encoded in the mitochondrial genome are part of which pathway?
3. What other molecules are encoded in the mitochondrial genome that are necessary
for protein synthesis?
4. Why do some antibiotics inhibit protein synthesis in mitochondria?
5. Why do mitochondria have many similarities to bacteria?
6. Why does mtDNA have a higher mutation rate than nuclear DNA?
6. How many copies of mtDNA are present in each mitochondrion? How many copies
are present in each cell?
8. Are all of the copies of mtDNA in an individual likely to be identical?
9. What is it called if a mitochondrion or a cell contains both normal and mutant
mtDNA?
10. What events can contribute to the presence of both normal and mutant mtDNA
in an individual?
11. What events can contribute to the presence of different levels of normal and
mutant mtDNA in different organs of an individual? What is this called?
12. Define the term mutational load and describe why it is important in
mitochondrial disease.
13. What two factors determine if there will be clinical expression of a disease
caused by mutation in mtDNA?
14. How do mutations in mtDNA cause detrimental effects on mitochondrial
function?
15.

What symptoms are likely to arise from mutations in mtDNA in skeletal muscle?

16. What symptoms are likely to arise from mutations in mtDNA in the central
nervous system?
17. Why are cardiac muscle, skeletal muscle and the central nervous system most
sensitive to mutation in mtDNA?
18. What are Lewy bodies and what role do they play in development of Parkinsons
disease?
19. What component of the electron transfer chain is inhibited by rotenone or MPP+
and leads to Parkinson-like symptoms?
20. What protein is always found in the Lewy bodies?