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Flow cytometry based techniques to study testicular acidophilic granulocytes from the protandrous fish gilthead seabream (Sparus aurata L.)
Biological Procedures Online volume 6, pages 129–136 (2004)
Abstract
The gilthead seabream is a protandrous seasonal breeding teleost that is an excellent model for studying the testicular regression process which occurs in both seasonal testicular involution and sex reversion. Little is known about the cell types and the molecular mechanisms involved in such processes, mainly because of the lack of appropriate methods for testis dissociation, and testicular cell isolation, culture and functional characterization. We have previously reported that gilthead seabream acidophilic granulocytes infiltrate the testis at post-spawning stage, settle close to the spermatogonia and accumulate intracellular interleukin-1β. In this paper, we report several flow cytometry based assays which allow to establish the role played by gilthead seabream testicular acidophilic granulocytes and permits their quantification.
Abbreviations
- BW:
-
body weight
- BSA:
-
bovine serum albumin
- SR-2:
-
steroid-free synthetic serum
- PBS:
-
phosphate-buffered saline
- FCS:
-
fetal calf serum
- FITC:
-
fluorescein isothiocyanate
- FSC:
-
forward scatter
- SSC:
-
side scatter
- TSB:
-
trypticase soy broth
- FC:
-
flow cytometry
- ANOVA:
-
one-way analysis of variance
- mAb:
-
monoclonal antibody
- IL-1β:
-
interleukin-1β
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Published: June 29, 2004.
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Chaves-Pozo, E., Mulero, V., Meseguer, J. et al. Flow cytometry based techniques to study testicular acidophilic granulocytes from the protandrous fish gilthead seabream (Sparus aurata L.). Biol. Proced. Online 6, 129–136 (2004). https://doi.org/10.1251/bpo81
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DOI: https://doi.org/10.1251/bpo81