Characterization of goldfish fin cells in culture: some evidence of an epithelial cell profile.
Comprehensive characterization of cultured cells in fish was little explored and cell origin is often deduced from morphological analogies with either epithelial of fibroblastic cells. This study aims to characterize cell origin in goldfish fin culture using morphological, immunochemical, and molecular approaches. Time lapse analysis revealed that cultured cell morphology changed within minutes. Therefore, cell morphology cannot predict whether cells are from fibroblastic or epithelial origin. The labeling pattern of heterologous anti-cytokeratin and anti-vimentin antibodies against goldfish epithelial cells and fibroblasts was first tested on skin sections and the corresponding labeling of the cultured cells was analyzed. No cell origin specificity could be obtained with the chosen antibodies. In the molecular approach, detection levels of three cytokeratin (CauK8-IIS, CauK49-IE and CauK50-Ie) and one vimentin transcripts were assessed on skin and fin samples. Specificity for epithelial cells of the most abundant mRNA, CauK49-Ie, was thereafter validated on skin sections by in situ hybridization. The selected markers were used afterwards to characterize fin cultures. CauK49-IE riboprobe labeled every cell in young cultures whereas no labeling was observed in older cultures. Accordingly, CauK49-IE transcript levels decreased after 15 days culture while CauK8-IIS ones increased. The use of homologous marker gave evidence that young cultured cells from goldfish fin are homogeneously of epithelial type and that cell characteristics may change over culture time.
Mauger
P-E
PE
Institut National de la Recherche Agronomique, UR 1037 SCRIBE, Campus de Beaulieu, Rennes cedex, France.
Labbé
C
C
Bobe
J
J
Cauty
C
C
Leguen
I
I
Baffet
G
G
Le Bail
P-Y
PY
D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D019070:Cell Lineage; D048430:Cell Shape; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology; D029941:Fish Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006054:Goldfish / Q000033:anatomy & histology*; D007150:Immunohistochemistry; D007633:Keratins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000166:cytology; D014158:Transcription, Genetic / Q000235:genetics; D014746:Vimentin / Q000235:genetics
D015415:Biomarkers; D029941:Fish Proteins; D012333:RNA, Messenger; D014746:Vimentin; D007633:Keratins