Fluoresceinated peanut agglutinin (PNA) is a marker for live O(2) sensing glomus cells in rat carotid body.
Experiments using live dissociated carotid body (CB) cells for patch clamping, [Ca(++)](i) or other measurements require positive identification of the cell being recorded. At present, cell morphology is usually employed, but several cell types within the carotid body evidence similar morphologic characteristics. Therefore, we sought to develop a method utilizing a vital dye to identify glomus cells before and during experiments that require live cells, such as patch clamp studies. It was previously reported that the binding sites for peanut agglutinin (PNA) were highly expressed by all neuroendocrine-derivatives of the sympathoadrenal neural crest, including glomus cells, small, intensely fluorescent cells, PC-12 cells, and adrenal chromaffin cells in situ (katz et al. 1995). By utilizing the binding characteristics of galactose-specific lectin peanut agglutinin (PNA) on the outer cell membrane, we tested the possibility that the fluoresceinated PNA may preferentially bind to CB glomus cells. The results to date show: (1) Rhodamine tagged PNA (Rhod-PNA) binds to the live dissociated glomus cells in less than one hour incubation and can be visualized in superfused cells; (2) Rhod-PNA labeled cells are perfectly matched with tyrosine hydroxylase (TH) positive glomus cells; (3) Rhod-PNA did not interfere with Fura-2 for Ca(++) imaging; (4) Rhod-PNA bound to glomus cells in [Ca(++)](i) studies does not affect O(2) response of glomus cells. Thus fluoresceinated PNA may be a useful marker for live CB glomus studies, without adversely affecting their physiologic response.
Kim
I
I
Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA. kiminsook@uams.ed
Yang
D J
DJ
Donnelly
D F
DF
Carroll
J L
JL
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002344:Carotid Body / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D002470:Cell Survival; D019793:Fluorescein / Q000378:metabolism*; D066298:In Vitro Techniques; D010100:Oxygen / Q000378:metabolism*; D019887:Peanut Agglutinin / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling
D015415:Biomarkers; D019887:Peanut Agglutinin; D010100:Oxygen; D019793:Fluorescein