The glucagon-like
peptide-1 receptor (GLP1R) is expressed in peripheral
tissues and the brain, where it exerts pleiotropic actions on metabolic
and inflammatory processes. Detection and visualization of GLP1R remains
challenging, partly due to a lack of validated reagents. Previously,
we generated
LUXendins
, antagonistic red and far-red
fluorescent probes for specific labeling of GLP1R in live and fixed
cells/tissues. We now extend this concept to the green and near-infrared
color ranges by synthesizing and testing
LUXendin492
,
LUXendin551
,
LUXendin615
, and
LUXendin762
. All four probes brightly and specifically label GLP1R in cells
and pancreatic islets. Further,
LUXendin551
acts as a
chemical beta cell reporter in preclinical rodent models, while
LUXendin762
allows noninvasive imaging, highlighting differentially
accessible GLP1R populations. We thus expand the color palette of
LUXendins
to seven different spectra, opening up a range of
experiments using wide-field microscopy available in most labs through
super-resolution imaging and whole animal imaging. With this, we expect
that
LUXendins
will continue to generate novel and specific
insights into GLP1R biology.
The glucagon-like peptide-1 receptor (GLP1R) is expressed in peripheral tissues and the brain, where it exerts pleiotropic actions on metabolic and inflammatory processes. Detection and visualization of GLP1R remains challenging, partly due to a lack of validated reagents. Previously, we generated LUXendins, antagonistic red and far-red fluorescent probes for specific labeling of GLP1R in live and fixed cells/tissue. We now extend this concept to the green and near-infrared color ranges by synthesizing and testing LUXendin492, LUXendin551, LUXendin615 and LUXendin762. All four probes brightly and specifically label GLP1R in cells and pancreatic islets. Further, LUXendin551 acts as chemical beta cell reporter in preclinical rodent models, while LUXendin762 allows non-invasive imaging, highlighting differentially-accessible GLP1R populations. We thus expand the color palette of LUXendins to seven different spectra, opening up a range of experiments using widefield microscopy available in most labs through super-resolution imaging and whole animal imaging. With this, we expect that LUXendins will continue to generate novel and specific insight into GLP1R biology.
Sulfonated rhodamines that endow xanthene dyes with cellular impermeability are presented. We fuse charged sulfonates to red and far-red dyes to obtain Sulfo549 and Sulfo646, respectively, and further link these to SNAP- and Halo-tag substrates for protein self-labelling. Cellular impermeability is validated in live cell imaging experiments in transfected HEK cells and neurons derived from induced pluripotent stem cells (iPSCs). Lastly, we show that Sulfo646 is amenable to STED nanoscopy by recording membranes of SNAP/Halo-surface-labelled human iPSC-derived neuronal axons. We therefore provide an avenue for rendering dyes impermeable for exclusive extracellular visualization via self-labelling protein tags.
To examine the influence of cellular immunity to spermatozoa on the fertility of inbred mice, leucocytes were transferred from other female mice that had been immunized with spermatozoa. Both syngeneic and allogeneic systems were investigatd. The cellular immune reaction was estimated using a footpad swelling test before and after the cell transfer. There was a significant increase of the paw thickness and a statistically significant reduction in fertility only for C3H mice immunized with C57BL spermatozoa.
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