Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration:
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2022
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| 100 | 1 | |a Müller, Stephan |d 1990- |e Verfasser |0 (DE-588)1287818552 |4 aut | |
| 245 | 1 | 0 | |a Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |c vorgelegt von Stephan Müller |
| 264 | 1 | |a Münster |c 2022 | |
| 300 | |a 128 Blätter |b Illustrationen, Diagramme |c 31 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Westfälische Wilhelms-Universität Münster |d 2023 |g Beschränkt für den Austausch | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1287695345/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034922387&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-034922387 | ||
| 883 | 2 | |8 1\p |a dnb |d 20230810 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
| 883 | 2 | |8 2\p |a dnb |d 20230810 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
Datensatz im Suchindex
| _version_ | 1804186439810809856 |
|---|---|
| adam_text | TABLE
OF
CONTENT
1.
INTRODUCTION
1
1.1
METABOLIC
PATHWAYS
ARE
DYNAMICALLY
REGULATED
AND
INTERCONNECTED
1
1.1.1
UNDERSTANDING
METABOLISM
DYNAMICS
1
1.1.2
REACTIVE
OXYGEN
SPECIES
AND
THEIR
PRESUMED
INFLUENCE
ON
GLUCOSE
METABOLISM
2
1.1.3
THE
PENTOSE-PHOSPHATE-PATHWAY
PLAYS
A
MAJOR
ROLE
IN
CLEARING
REACTIVE
OXYGEN
SPECIES
4
1.1.4
THE
DROSOPHILA
CENTRAL
NERVOUS
SYSTEM
7
1.1.5
GLUCOSE
TRANSPORT
AND
GLUCOSE
RE-ROUTING
UPON
ALTERNATIVE
ENERGY
DEMAND
8
1.1.6
ASTROCYTE-NEURON-LACATE-SHUTTLE
HYPOTHESIS
9
1.2
LIVE
IMAGING
TO
UNRAVEL
METABOLITE
LEVELS
AND
DYNAMICS
IN
LIVING
TISSUES
1.2.1
GENETICALLY
ENCODED
FLUORESCENT
METABOLITE
INDICATORS
10
1.2.2
USING
SF-SENSORS
WITH
AN
INSENSITIVE
CONTROL
11
1.2.2.1
METABOLISM
INDUCED
CHANGES
IN
PH
CAN
AFFECT
SINGLE
FLUOROPHORE
SENSOR
IMAGING
12
1.2.2.2
INAP1
IS
A
NADPH
SF-SENSOR
13
1.2.2.3
HYPERRED
IS
A
H2O2
SF-SENSOR
14
1.2.2.4
SENSORS
NEED
TO
BE
CALIBRATED
FOR
EVERY
MODEL
14
1.3
AIMS
OF
THIS
STUDY
17
2.
MATERIALS
AND
METHODS
18
2.1
MATERIALS
18
2.1.1
FOOD
AND
MEDIA
18
2.1.2
KITS
22
2.1.3
OLIGONUCLEOTIDES
22
2.1.4
PLASMIDS
23
2.2
DROSOPHILA
METHODS
24
2.2.1
FLY
STRAINS,
HUSBANDRY,
AND
DROSOPHILA
METHODS
24
2.2.2
DROSOPHILA
PREPARATIONS
27
2.2.2.1
LARVAL
AND
ADULT
BRAIN
DISSECTIONS
FOR
STAINING
ANDLIVE
IMAGING
27
2.2.2.2
ANTIBODY
STAINING
28
2.2.2.3S
EMI-THIN-SECTIONS
28
2.2.2.4
GERMLINE
TRANSFORMATION
28
2.2.2.5
LIFE
SPAN
ANALYSIS
29
2.3
MOLECULAR
BIOLOGY
30
2.3.1
Q5
PCR
30
2.3.2
DNA
GEL
ELEKTROPHORESES
30
2.3.3
GEL
FRAGMENT
AND
PCR
PRODUCT
PURIFICATION
30
2.3.4
PLASMID
DNA
PREPARATION,
MEASUREMENTS,
AND
SEQUENCING
30
2.3.5
MOLECULAR
CLONING
31
2.3.6
RNA
EXTRACTION
AND
RTPCR
32
2.4
MICROSCOPY,
LIVE
IMAGING,
AND
ANALYSIS
32
2.4.1
GENERAL
MICROSCOPY
SETTINGS
32
2.4.2
LIVE
IMAGING
33
2.4.3
ANALYSIS
33
3.
RESULTS
35
3.1
THE
IMPORTANCE
OF
THE
PENTOSE-PHOSPHATE-PATHWAY
DIFFERS
BETWEEN
TISSUE
TYPES
35
3.1.1
GENERAL
DOWN-REGULATION
OF
THE
PENTOSE-PHOSPHATE-PATHWAY
THROUGHOUT
DEVELOPMENT
CAUSES
LETHALITY,
TISSUE-SPECIFIC
DOWN-REGULATING
DOES
NOT
35
3.1.2
LIFE
SPAN
ANALYSIS
IS
NOT
ENOUGH
TO
DISSECT
A
SPECIFIC
ROLE
OF
THE
PENTOSE
PHOSPHATE-PATHWAY
IN
AGING
FLIES
36
3.2
PAN-NEURONAL,
BUT
ALSO
PAN-GLIAL
KNOCK-DOWN
OF
THE
PENTOSE-PHOSPHATE-PATHWAY
CAN
CAUSE
PROGRESSIVE
NEURODEGENERATION
40
3.2.1
SEMI-THIN-SECTIONS
REVEAL
NEURONAL
CELL
LOSS
UPON
OXIDATIVE
PENTOSE
PHOSPHATE-PATHWAY
REDUCTION
40
3.2.2
ANTIBODY
STAINING
OF
ADULT
BRAINS
ALLOWED
TO
REVEAL
ACTIVITY-DEPENDENT
INCREASE
IN
NEURODEGENERATION
46
3.2.3
ANTIOXIDANT
TREATMENT
REVERSES
THE
DEGENERATIVE
PHENOTYPE
CAUSED
BY
PAN
NEURONAL
KNOCK
DOWN
OF
G6PDH
AND
6-PGD
IN
LIGHT/DARKNESS
CONDITIONS
48
3.3
METABOLIC
LIVE-IMAGING
ELUCIDATES
RELATIONSHIP
BETWEEN
ALTERED
NADPH
LEVELS
AND
INCREASING
H2O2
58
3.3.1
CHARACTERIZATION
OF
INAP1
AND
HYPERRED
SHOWS
TARGET
BINDING
58
AND
REVERSIBILITY
3.3.2
PH-MEASUREMENTS
CONFIRM
FEASIBILITY
OF
SINGLE
FLUOROPHORE
61
IMAGING
APPROACH
3.3.3
ALTERED
NADPH
LEVELS
FOLLOWING
DOWN-REGULATION
OF
OXPPP
CAN
BE
SEEN
AFTER
FLUORESCENCE
RECOVERY
64
3.3.4
H202-MEASUREMENTS
EXHIBIT
HIGHER
LEVELS
FOLLOWING
A
NADPH-REDUCTION
UPON
DOWN-REGULATION
OF
THE
OXIDATIVE
BRANCH
OF
THE
PENTOSE-PHOSPHATE
PATHWAY
67
3.3.5
SIMULTANEOUS
NADPH
AND
H202-MEASUREMENTS
IN
ADULT
DROSOPHILA
BRAINS
SHOW
PARTLY
DECREASED
NADPH
LEVELS
AND
A
WELL
BUFFERED
H2O2
SYSTEM
70
4.
DISCUSSION
72
4.1
TRANSIENT
METABOLIC
DISTURBANCES
LEAD
TO
SLIGHTLY
ALTERED
LIFE
EXPECTANCY,
BUT
HINT
ON
WELL
BALANCED
REDOX-CONTROL
70
4.2
SYSTEMIC
ACTION
OF
BROADLY
EXPRESSED
ROS
AND
NADPH
COMPARTMENTALIZATION
DETERMINE
THE
NEED
FOR
ROBUST
READOUT
METHODS
TO
UNRAVEL
SITE
SPECIFIC
EFFECTS
75
4.3
PROLONGED
REDUCTION
OF
NADPH
PRODUCING
ENZYMES
INCREASES
AGE
AND
ACTIVITY
DEPENDENT
NEURODEGENERATION
76
4.4
BASIC
FEATURES
OF
BIOSENSORS
DETERMINE
THE
NEED
FOR
ELABORATED
NORMALIZATION
PROTOCOLS
78
4.5
NADPH
LEVELS
DIFFER
AS
H2O2
LEVELS
INCREASE
UPON
OXPPP
REDUCTION
IN
DROSOPHILA
EYEDISCS
79
4.6
ADULT
BRAIN
MULTI-PARAMETRIC
LIVE
IMAGING
SHOWS
NADPH
REDUCTION
IN
A
WELL
BUFFERED
H2O2
BACKGROUND
81
5.
SUMMARY
83
6.
REFERENCES
84
7.
APPENDIX
107
7.1
LIST
OF
FIGURES
107
7.2
LIST
OF
ABBREVIATIONS
108
7.3
EXEMPLARY
R-CODE
110
7.3.1
LIFE
SPAN
ANALYSIS
110
7.3.2
NEURODEGENERATION
111
7.3.3
PHERRY
AND
INAP
ANALYSIS
113
7.3.4
HYPER
ANALYSIS
118
7.4
SUPPLEMENTARY
FIGURES
123
LEBENSLAUF
126
STATEMENT
OF
CONFIDENCE
127
ACKNOWLEDGMENTS
128
|
| adam_txt |
TABLE
OF
CONTENT
1.
INTRODUCTION
1
1.1
METABOLIC
PATHWAYS
ARE
DYNAMICALLY
REGULATED
AND
INTERCONNECTED
1
1.1.1
UNDERSTANDING
METABOLISM
DYNAMICS
1
1.1.2
REACTIVE
OXYGEN
SPECIES
AND
THEIR
PRESUMED
INFLUENCE
ON
GLUCOSE
METABOLISM
2
1.1.3
THE
PENTOSE-PHOSPHATE-PATHWAY
PLAYS
A
MAJOR
ROLE
IN
CLEARING
REACTIVE
OXYGEN
SPECIES
4
1.1.4
THE
DROSOPHILA
CENTRAL
NERVOUS
SYSTEM
7
1.1.5
GLUCOSE
TRANSPORT
AND
GLUCOSE
RE-ROUTING
UPON
ALTERNATIVE
ENERGY
DEMAND
8
1.1.6
ASTROCYTE-NEURON-LACATE-SHUTTLE
HYPOTHESIS
9
1.2
LIVE
IMAGING
TO
UNRAVEL
METABOLITE
LEVELS
AND
DYNAMICS
IN
LIVING
TISSUES
1.2.1
GENETICALLY
ENCODED
FLUORESCENT
METABOLITE
INDICATORS
10
1.2.2
USING
SF-SENSORS
WITH
AN
INSENSITIVE
CONTROL
11
1.2.2.1
METABOLISM
INDUCED
CHANGES
IN
PH
CAN
AFFECT
SINGLE
FLUOROPHORE
SENSOR
IMAGING
12
1.2.2.2
INAP1
IS
A
NADPH
SF-SENSOR
13
1.2.2.3
HYPERRED
IS
A
H2O2
SF-SENSOR
14
1.2.2.4
SENSORS
NEED
TO
BE
CALIBRATED
FOR
EVERY
MODEL
14
1.3
AIMS
OF
THIS
STUDY
17
2.
MATERIALS
AND
METHODS
18
2.1
MATERIALS
18
2.1.1
FOOD
AND
MEDIA
18
2.1.2
KITS
22
2.1.3
OLIGONUCLEOTIDES
22
2.1.4
PLASMIDS
23
2.2
DROSOPHILA
METHODS
24
2.2.1
FLY
STRAINS,
HUSBANDRY,
AND
DROSOPHILA
METHODS
24
2.2.2
DROSOPHILA
PREPARATIONS
27
2.2.2.1
LARVAL
AND
ADULT
BRAIN
DISSECTIONS
FOR
STAINING
ANDLIVE
IMAGING
27
2.2.2.2
ANTIBODY
STAINING
28
2.2.2.3S
EMI-THIN-SECTIONS
28
2.2.2.4
GERMLINE
TRANSFORMATION
28
2.2.2.5
LIFE
SPAN
ANALYSIS
29
2.3
MOLECULAR
BIOLOGY
30
2.3.1
Q5
PCR
30
2.3.2
DNA
GEL
ELEKTROPHORESES
30
2.3.3
GEL
FRAGMENT
AND
PCR
PRODUCT
PURIFICATION
30
2.3.4
PLASMID
DNA
PREPARATION,
MEASUREMENTS,
AND
SEQUENCING
30
2.3.5
MOLECULAR
CLONING
31
2.3.6
RNA
EXTRACTION
AND
RTPCR
32
2.4
MICROSCOPY,
LIVE
IMAGING,
AND
ANALYSIS
32
2.4.1
GENERAL
MICROSCOPY
SETTINGS
32
2.4.2
LIVE
IMAGING
33
2.4.3
ANALYSIS
33
3.
RESULTS
35
3.1
THE
IMPORTANCE
OF
THE
PENTOSE-PHOSPHATE-PATHWAY
DIFFERS
BETWEEN
TISSUE
TYPES
35
3.1.1
GENERAL
DOWN-REGULATION
OF
THE
PENTOSE-PHOSPHATE-PATHWAY
THROUGHOUT
DEVELOPMENT
CAUSES
LETHALITY,
TISSUE-SPECIFIC
DOWN-REGULATING
DOES
NOT
35
3.1.2
LIFE
SPAN
ANALYSIS
IS
NOT
ENOUGH
TO
DISSECT
A
SPECIFIC
ROLE
OF
THE
PENTOSE
PHOSPHATE-PATHWAY
IN
AGING
FLIES
36
3.2
PAN-NEURONAL,
BUT
ALSO
PAN-GLIAL
KNOCK-DOWN
OF
THE
PENTOSE-PHOSPHATE-PATHWAY
CAN
CAUSE
PROGRESSIVE
NEURODEGENERATION
40
3.2.1
SEMI-THIN-SECTIONS
REVEAL
NEURONAL
CELL
LOSS
UPON
OXIDATIVE
PENTOSE
PHOSPHATE-PATHWAY
REDUCTION
40
3.2.2
ANTIBODY
STAINING
OF
ADULT
BRAINS
ALLOWED
TO
REVEAL
ACTIVITY-DEPENDENT
INCREASE
IN
NEURODEGENERATION
46
3.2.3
ANTIOXIDANT
TREATMENT
REVERSES
THE
DEGENERATIVE
PHENOTYPE
CAUSED
BY
PAN
NEURONAL
KNOCK
DOWN
OF
G6PDH
AND
6-PGD
IN
LIGHT/DARKNESS
CONDITIONS
48
3.3
METABOLIC
LIVE-IMAGING
ELUCIDATES
RELATIONSHIP
BETWEEN
ALTERED
NADPH
LEVELS
AND
INCREASING
H2O2
58
3.3.1
CHARACTERIZATION
OF
INAP1
AND
HYPERRED
SHOWS
TARGET
BINDING
58
AND
REVERSIBILITY
3.3.2
PH-MEASUREMENTS
CONFIRM
FEASIBILITY
OF
SINGLE
FLUOROPHORE
61
IMAGING
APPROACH
3.3.3
ALTERED
NADPH
LEVELS
FOLLOWING
DOWN-REGULATION
OF
OXPPP
CAN
BE
SEEN
AFTER
FLUORESCENCE
RECOVERY
64
3.3.4
H202-MEASUREMENTS
EXHIBIT
HIGHER
LEVELS
FOLLOWING
A
NADPH-REDUCTION
UPON
DOWN-REGULATION
OF
THE
OXIDATIVE
BRANCH
OF
THE
PENTOSE-PHOSPHATE
PATHWAY
67
3.3.5
SIMULTANEOUS
NADPH
AND
H202-MEASUREMENTS
IN
ADULT
DROSOPHILA
BRAINS
SHOW
PARTLY
DECREASED
NADPH
LEVELS
AND
A
WELL
BUFFERED
H2O2
SYSTEM
70
4.
DISCUSSION
72
4.1
TRANSIENT
METABOLIC
DISTURBANCES
LEAD
TO
SLIGHTLY
ALTERED
LIFE
EXPECTANCY,
BUT
HINT
ON
WELL
BALANCED
REDOX-CONTROL
70
4.2
SYSTEMIC
ACTION
OF
BROADLY
EXPRESSED
ROS
AND
NADPH
COMPARTMENTALIZATION
DETERMINE
THE
NEED
FOR
ROBUST
READOUT
METHODS
TO
UNRAVEL
SITE
SPECIFIC
EFFECTS
75
4.3
PROLONGED
REDUCTION
OF
NADPH
PRODUCING
ENZYMES
INCREASES
AGE
AND
ACTIVITY
DEPENDENT
NEURODEGENERATION
76
4.4
BASIC
FEATURES
OF
BIOSENSORS
DETERMINE
THE
NEED
FOR
ELABORATED
NORMALIZATION
PROTOCOLS
78
4.5
NADPH
LEVELS
DIFFER
AS
H2O2
LEVELS
INCREASE
UPON
OXPPP
REDUCTION
IN
DROSOPHILA
EYEDISCS
79
4.6
ADULT
BRAIN
MULTI-PARAMETRIC
LIVE
IMAGING
SHOWS
NADPH
REDUCTION
IN
A
WELL
BUFFERED
H2O2
BACKGROUND
81
5.
SUMMARY
83
6.
REFERENCES
84
7.
APPENDIX
107
7.1
LIST
OF
FIGURES
107
7.2
LIST
OF
ABBREVIATIONS
108
7.3
EXEMPLARY
R-CODE
110
7.3.1
LIFE
SPAN
ANALYSIS
110
7.3.2
NEURODEGENERATION
111
7.3.3
PHERRY
AND
INAP
ANALYSIS
113
7.3.4
HYPER
ANALYSIS
118
7.4
SUPPLEMENTARY
FIGURES
123
LEBENSLAUF
126
STATEMENT
OF
CONFIDENCE
127
ACKNOWLEDGMENTS
128 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Müller, Stephan 1990- |
| author_GND | (DE-588)1287818552 |
| author_facet | Müller, Stephan 1990- |
| author_role | aut |
| author_sort | Müller, Stephan 1990- |
| author_variant | s m sm |
| building | Verbundindex |
| bvnumber | BV049577380 |
| ctrlnum | (OCoLC)1388272060 (DE-599)DNB1287695345 |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV049577380 |
| illustrated | Illustrated |
| index_date | 2024-07-03T23:31:35Z |
| indexdate | 2024-07-10T10:11:09Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034922387 |
| oclc_num | 1388272060 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 128 Blätter Illustrationen, Diagramme 31 cm |
| publishDate | 2022 |
| publishDateSearch | 2022 |
| publishDateSort | 2022 |
| record_format | marc |
| spelling | Müller, Stephan 1990- Verfasser (DE-588)1287818552 aut Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration vorgelegt von Stephan Müller Münster 2022 128 Blätter Illustrationen, Diagramme 31 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische Wilhelms-Universität Münster 2023 Beschränkt für den Austausch (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1287695345/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034922387&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p dnb 20230810 DE-101 https://d-nb.info/provenance/plan#dnb 2\p dnb 20230810 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Müller, Stephan 1990- Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |
| subject_GND | (DE-588)4113937-9 |
| title | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |
| title_auth | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |
| title_exact_search | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |
| title_exact_search_txtP | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |
| title_full | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration vorgelegt von Stephan Müller |
| title_fullStr | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration vorgelegt von Stephan Müller |
| title_full_unstemmed | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration vorgelegt von Stephan Müller |
| title_short | Alternative glucose metabolization prevents ROS-induced progressive neurodegeneration |
| title_sort | alternative glucose metabolization prevents ros induced progressive neurodegeneration |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1287695345/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034922387&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT mullerstephan alternativeglucosemetabolizationpreventsrosinducedprogressiveneurodegeneration |
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