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DTSTART:20241018T130000Z
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TRANSP:OPAQUE
LOCATION:Auditorium and Online (Teams)
SUMMARY:ICFO | JESSICA ANGULO CAPEL
CLASS:PUBLIC
DESCRIPTION:Intracellular trafficking\, particularly protein secretion\, fa
 ces numerous unresolved challenges. This thesis aims to provide and evalua
 te tools for quantitative investigation of these processes using fluoresce
 nt microscopy. Quantitative analysis offers two main benefits: detailed ch
 aracterization of molecular dynamics for mechanistic understanding and obj
 ective measurements for accurate comparisons across experiments.\nIn Chapt
 er 1\, we introduce the secretory pathway\, a cellular pathway responsible
  for the synthesis\, processing\, sorting and delivery of secretory protei
 ns to the extracellular environment. In Chapter 2\, we provide a thorough 
 description of the methodologies used in this thesis. They include various
  fluorescence microscopy techniques\, automated image analysis\, and biolo
 gical methods tailored to the secretory pathway. The tools were selected t
 o achieve high spatial and temporal resolution\, enable quantitative analy
 sis\, and allow live-cell characterization.\nIn Chapter 3\, we used fluore
 scence imaging to objectively evaluate results in four projects addressing
  protein secretion and intracellular trafficking. These included quantifyi
 ng colocalization and proximity of structures\, measuring fluorescent inte
 nsity differences\, and characterizing dynamics of particles like ERGIC-de
 rived nanotubules. Consistent sample preparation and image acquisition\, c
 oupled with computational analysis\, are crucial for accurate\, unbiased r
 esults.\nChapter 4 focuses on single-particle tracking (SPT) in the secret
 ory pathway\, proposing control experiments and parameter descriptors to m
 aximize data quality. We emphasized labeling strategies\, imaging\, and da
 ta analysis considerations for reliable results.\nChapter 5 applied these 
 methodologies to study protein sorting at the TGN\, examining the role of 
 ER-Golgi membrane contact sites (MCS) in TGN-derived carrier biogenesis. U
 sing super-resolution fluorescence microscopy\, we identified cargo accumu
 lation regions and conducted SPT experiments\, revealing confined\, slow m
 otion of cargo proteins near MCS. This effect was inhibited by the lipid t
 ransfer blocker 25-HC\, indicating upstream regulation of cargo localizati
 on preferences by MCS.\n&nbsp\;\nFriday October 10\, 15:00 h. ICFO Auditor
 ium \nThesis Director: Prof. Dr. Mar&iacute\;a Garc&iacute\;a-Parajo and D
 r. Felix Campelo Aubarell
DTSTAMP:20260407T073753Z
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