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2015 ; 10
(10
): e0139938
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Transcriptomic Analysis of Trout Gill Ionocytes in Fresh Water and Sea Water
Using Laser Capture Microdissection Combined with Microarray Analysis
#MMPMID26439495
Leguen I
; Le Cam A
; Montfort J
; Peron S
; Fautrel A
PLoS One
2015[]; 10
(10
): e0139938
PMID26439495
show ga
Fish gills represent a complex organ composed of several cell types that perform
multiple physiological functions. Among these cells, ionocytes are implicated in
the maintenance of ion homeostasis. However, because the ionocyte represents only
a small percent of whole gill tissue, its specific transcriptome can be
overlooked among the numerous cell types included in the gill. The objective of
this study is to better understand ionocyte functions by comparing the RNA
expression of this cell type in freshwater and seawater acclimated rainbow trout.
To realize this objective, ionocytes were captured from gill cryosections using
laser capture microdissection after immunohistochemistry. Then, transcriptome
analyses were performed on an Agilent trout oligonucleotide microarray. Gene
expression analysis identified 108 unique annotated genes differentially
expressed between freshwater and seawater ionocytes, with a fold change higher
than 3. Most of these genes were up-regulated in freshwater cells. Interestingly,
several genes implicated in ion transport, extracellular matrix and structural
cellular proteins appeared up-regulated in freshwater ionocytes. Among them,
several ion transporters, such as CIC2, SLC26A6, and NBC, were validated by qPCR
and/or in situ hybridization. The latter technique allowed us to localize the
transcripts of these ion transporters in only ionocytes and more particularly in
the freshwater cells. Genes involved in metabolism and also several genes
implicated in transcriptional regulation, cell signaling and the cell cycle were
also enhanced in freshwater ionocytes. In conclusion, laser capture
microdissection combined with microarray analysis allowed for the determination
of the transcriptional signature of scarce cells in fish gills, such as
ionocytes, and aided characterization of the transcriptome of these cells in
freshwater and seawater acclimated trout.