Abstract
Although copper has been reported to influence numerous proteins known to be important for angiogenesis, the enhanced sensitivity of this developmental process to copper bioavailability has remained an enigma, because copper metalloproteins are prevalent and essential throughout all cells. Recent developments in x-ray optics at third-generation synchrotron sources have provided a resource for highly sensitive visualization and quantitation of metalloproteins in biological samples. Here, we report the application of x-ray fluorescence microscopy (XFM) to in vitro models of angiogenesis and neurogenesis, revealing a surprisingly dramatic spatial relocalization specific to capillary formation of 80-90% of endogenous cellular copper stores from intracellular compartments to the tips of nascent endothelial cell filopodia and across the cell membrane. Although copper chelation had no effect on process formation, an almost complete ablation of network formation was observed. XFM of highly vascularized ductal carcinomas showed copper clustering in putative neoangiogenic areas. This use of XFM for the study of a dynamic developmental process not only sheds light on the copper requirement for endothelial tube formation but highlights the value of synchrotron-based facilities in biological research.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2247-2252 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 104 |
| Issue number | 7 |
| DOIs | |
| State | Published - Feb 13 2007 |
| Externally published | Yes |
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Keywords
- Copper chelation
- Human microvascular endothelial cells
- Infiltrating ductal breast carcinoma
ASJC Scopus subject areas
- Genetics
- General
Cite this
X-ray flourescence microscopy reveals large-scale relocalization and extracellular translocation of cellular copper during angiogenesis. / Finney, Lydia; Mandava, Suneeta; Ursos, Lyann; Zhang, Wen; Rodi, Diane; Vogt, Stefan; Legnini, Daniel; Maser, Jorg; Ikpatt, Offiong; Olopade, Olyfunmilayo I.; Glesne, David.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 7, 13.02.2007, p. 2247-2252.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - X-ray flourescence microscopy reveals large-scale relocalization and extracellular translocation of cellular copper during angiogenesis
AU - Finney, Lydia
AU - Mandava, Suneeta
AU - Ursos, Lyann
AU - Zhang, Wen
AU - Rodi, Diane
AU - Vogt, Stefan
AU - Legnini, Daniel
AU - Maser, Jorg
AU - Ikpatt, Offiong
AU - Olopade, Olyfunmilayo I.
AU - Glesne, David
PY - 2007/2/13
Y1 - 2007/2/13
N2 - Although copper has been reported to influence numerous proteins known to be important for angiogenesis, the enhanced sensitivity of this developmental process to copper bioavailability has remained an enigma, because copper metalloproteins are prevalent and essential throughout all cells. Recent developments in x-ray optics at third-generation synchrotron sources have provided a resource for highly sensitive visualization and quantitation of metalloproteins in biological samples. Here, we report the application of x-ray fluorescence microscopy (XFM) to in vitro models of angiogenesis and neurogenesis, revealing a surprisingly dramatic spatial relocalization specific to capillary formation of 80-90% of endogenous cellular copper stores from intracellular compartments to the tips of nascent endothelial cell filopodia and across the cell membrane. Although copper chelation had no effect on process formation, an almost complete ablation of network formation was observed. XFM of highly vascularized ductal carcinomas showed copper clustering in putative neoangiogenic areas. This use of XFM for the study of a dynamic developmental process not only sheds light on the copper requirement for endothelial tube formation but highlights the value of synchrotron-based facilities in biological research.
AB - Although copper has been reported to influence numerous proteins known to be important for angiogenesis, the enhanced sensitivity of this developmental process to copper bioavailability has remained an enigma, because copper metalloproteins are prevalent and essential throughout all cells. Recent developments in x-ray optics at third-generation synchrotron sources have provided a resource for highly sensitive visualization and quantitation of metalloproteins in biological samples. Here, we report the application of x-ray fluorescence microscopy (XFM) to in vitro models of angiogenesis and neurogenesis, revealing a surprisingly dramatic spatial relocalization specific to capillary formation of 80-90% of endogenous cellular copper stores from intracellular compartments to the tips of nascent endothelial cell filopodia and across the cell membrane. Although copper chelation had no effect on process formation, an almost complete ablation of network formation was observed. XFM of highly vascularized ductal carcinomas showed copper clustering in putative neoangiogenic areas. This use of XFM for the study of a dynamic developmental process not only sheds light on the copper requirement for endothelial tube formation but highlights the value of synchrotron-based facilities in biological research.
KW - Copper chelation
KW - Human microvascular endothelial cells
KW - Infiltrating ductal breast carcinoma
UR - http://www.scopus.com/inward/record.url?scp=33847775171&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33847775171&partnerID=8YFLogxK
U2 - 10.1073/pnas.0607238104
DO - 10.1073/pnas.0607238104
M3 - Article
C2 - 17283338
AN - SCOPUS:33847775171
VL - 104
SP - 2247
EP - 2252
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 7
ER -