Oligodendrocyte-derived exosome (continued)

{Dutta, 2021 #1542} PD & MSA, HC,

Plasma collection Plasma samples were obtained as described previously [2]. Briefly, peripheral blood was drawn by venipuncture and collected into EDTA tubes. → Within 30 min of collection, the plasma tubes underwent centrifugation at 4 °C for 15 min at 1500g. → The plasma then was aliquoted into 0.5 mL aliquots (control samples) or 1.0 mL aliquots (MSA samples) and stored at − 80 °C.

Postmortem serum was obtained as described previously [5]. Briefly, blood was drawn from the left ventricle by a transthoracic puncture using 30-mL, disposable, polyethylene syringes fitted with 8-cm long, 18-gauge needles. → Serum was separated from the blood using standard separator vacuum tubes (7 mL) → prior to 10 min centrifugation, → aliquoted into 0.5-mL polyethylene microcentrifuge tubes, and stored at − 80 °C.

Eye

• general good review: 2012 London, Sung

Fibroblast

• Gene
  • Maintain genetic background
• Origin
  • Mesenchyme, ie mesoderm
• Function
  • to secrete extracellular matrix, ( eg. collagen) which forms the connective tissue of an organ
  • Synthesis of extracellular matrix (ECM), epithelial differentiation, regulation of inflammation and wound healing (Tarin and Croft 1969). In addition, fibroblasts are also responsible for secretion of growth factors and work as scaffolds for several other cell types, acting as the main mediator cell for tissue fibrosis and scar formation (Parsonage et al. 2005; Tomasek et al. 2002).
• Structure
  • Cytoskeleton
    • Microtubule
    • Actin
    • Intermediate filiament
      • Vimentin
        • Major component of intermediate filament of fibroblast
• Mitochondria
  • Dynamic is a little different
  • Fission, complex I, basal mitochondrial membrane potential (Ψm)
  • similarities in mitochondrial dysfunction between fibroblasts and neurons^{2,3,4,5,6,7,8,9,10,11}
• Culture
  • In tissue culture, human fibroblasts can survive up to 50 generations (Hayflick and Moorhead, 1961).
  • The ensuing cell culture is a mixture of primary fibroblasts and keratinocytes at the beginning of the culturing process and a pure culture of fibroblasts is only achieved in the third passage. However, the fibroblast population consists most probably of a mixture of mitotic and postmitotic fibroblast [3], thus contributing to a heterogeneous cell population even at early passages
  • patient skin fibroblasts can be obtained repeatedly with ease and thus might be used even to monitor disease progression,
  • amenable to genetic manipulation via electroporation or lentiviral constructs, KD ( si RNA)
  • passage number
    • should be matched between assays, between group?
    • <10 (Cartelli et al. 2012, PMID 22666358)

Cell model system for CNS?

		pros	Cons
Maintain genetic background Cell physiology can be, mitophagy, autophagy Cell organelles, mitochondira, lysosome Express, PARKIN, pink1, GBA Recapitulates dysfunction of GBA, mitophagy			Hardly express aSyn 없지. neuron 아니므로 Axonal transport 못 보나 본데?
To identify markers that
Gene regulation /activation
Epigenetic modification
Histone ubiquitination
Mi RNA expression
Cell physiology	Mito metabolism
	Cytoskeleton: -	[cytoskeleton] the levels of α-tubulin, b-tubulin, actin, vimentin (by wb), (Cartelli et al. 2012, PMID 22666358) FIG 2 [microtubule stability] tubulin PTM: Tubulin acetylation/tyrosination/detyrosination: fig3 (WB) MT mass (?): the amount of α-tubulin associated with Triton-soluble, i.e. dimeric pool (Dim), and with Triton-insoluble fraction, polymerized MT fraction (MT). Altered in PD → corrected by taxol	Axon 은 없지. neuron 아니므로 Axonal transport 못 보나 본데?
	Lysosomal autophagy
Cell pathology

• Commercial source
  • Lifeline https://www.lifelinecelltech.com/fibroblasts-oh-the-places-you-will-go/
  • Cell Applications https://www.cellapplications.com/human-dermal-fibroblasts-hdf

GABA

BAMAN (?)	Efficacy modeling
		Sno Sensitive parameter Literature value (Source) value that used initially Value After optimization 1 Imax_GABA_StrD2_Gpe 64 fmol/mg (autoradiography in human cadavers) 0.12 umoles/Gpe 0.99 2 Imax_GABA_Gpe_STN 120 fmol/mg protein (autoradiography using flunazenam in monkey) 0.03 umoles/STN 0.9 3 Imax_GABA_StrD1_Gpi 43 fmol/mg (autoradiography in human cadavers) 0.03 umoles/Gpi 0.9 4 Smax_DA_SNc_StrD1 251.1 fmol/mg protein (autoradiography in human cadavers) 1.81 umoles/StrD1 0.68 5 Imax_DA_SNc_StrD2 216.8 fmol/mg protein (autoradiography in human cadavers) 1.18 moles/StrD2 0.4

Further readings

4. Lang AE, Lozano AM. Parkinson’s disease. First of two parts. N Engl J Med 1998;339:1044-53.
   
5. Lang AE, Lozano AM. Parkinson’s disease. Second of two parts. N Engl J Med 1998;339:1130-43.

GABA imaging

Flumaz {Kim, 2017 #1820}

[18F]flumazenil A novel tracer for PET imaging of human benzodiazepine receptors [18F]-flumazenil (FMZ) has been synthesized by various strategies involving complex purification by on-line high-performance liquid chromatography (HPLC) and also by solid-phase cartridges. [18F]-FMZ has been used for the assessment of the gamma-aminobutyric acid (GABA) receptors by positron emission tomography (PET).

{ng, 2019 #1821}

In MPTP monkey: ↑ GABA by HPLC, =GABA by MRS No }[18F]flumazenil PET in PD patients?

review {Magrinelli, 2016 #1063}

	early	Late
Mechanism	early PD, hypokinetic gait, which is characterized by reduced gait speed and amplitude with nearly normal cadence, is an expression of the bradykinesia	In later PD, different neurotransmitters are hypothesized to contribute to gait disturbance, including noradrenalin and serotonin systems, but converging evidence points to a key role of cholinergic dysfunction in the PPN (pedunculopontine nucleus) [21, 42, 43, 94]. Loss of cholinergic neurons in the PPN and NBM may contribute to posture and gait signs and falls through failure in the direct control of spinal circuitries and the deficits in the attentional processes required for these tasks [42, 43] and to cognitive impairment [7, 43].

Uncertain Spans

location	transcription	uncertainty
Cell model system / Hardly express aSyn / Axon	the right Cons cell shows both Hardly express aSyn and the bottom-row Korean note Axon 은 없지. neuron 아니므로 / Axonal transport 못 보나 본데?; alignment between rows is inferred.	matrix density.
GABA / BAMAN (?)	left column header reads BAMAN (sic) — possibly MAN or a mis-alignment of BA (left column) and MAN heading; preserved verbatim.	left edge clip.
GABA / Efficacy modeling / table 5 row Imax_DA_SNc_StrD2	initial value 1.18 moles/StrD2 is missing the u prefix that appears elsewhere as umoles; preserved verbatim.	source typo or OCR glitch — visible as moles not umoles.