Expression of Crystallins, Pax6, Filensin, CP49, MIP, and MP20 in lens-derived cell lines.
OBJECTIVE
Cell lines are the systems of choice to analyze cellular functions related to the particular organ system. For lens research, three cell lines are widely used: N/N1003A (derived from rabbit lenses), alpha TN4, and NKR-11 (both of murine origin). The aim of the current study was to characterize these particular cell lines with respect to their expression of genes that are considered to be lens specific or expressed preferentially in the lens, such as crystallins, Pax6, Filensin, CP49, MIP, and MP20.


METHODS
alpha A- and alpha B-crystallin cDNA from rabbit lenses were sequenced. The expression of various genes was analyzed by reverse transcription-polymerase chain reaction using specific primers and mRNA from three lens-derived cell lines. For control, the expression of the selected genes was compared in nonlenticular tissues of mouse as well as in non-lens-derived murine cell lines (EF43, NIH-3T3, and L929).


RESULTS
None of the transcripts for beta B2-crystallin, gamma-crystallins, MIP, MP20, filensin, and CP49 could be detected in the lens-derived cell lines. Transcripts for alpha A-crystallin were amplified in alpha TN4, but not in N/N1003A and NKR-11 cells. Pax6, a master control gene of eye development, is expressed in all three lens-derived cell lines and, additionally, in cell lines of neuronal origin, but not in corneal endothelial cells and in the currently used control cell lines.


CONCLUSIONS
Three cell lines of lenticular origin were tested for expression of genes that were found abundantly in the lens. The observed expression of Pax6 in all lens-derived cell lines allows their use in the analysis of corresponding signal chains.
Krausz
E
E
Institut für Säugetiergenetik, GSF-Forschungzentrum für Umwelt und Gesundheit, Neuherberg, Germany.
Augusteyn
R C
RC
Quinlan
R A
RA
Reddan
J R
JR
Russell
P
P
Sax
C M
CM
Graw
J
J
D000595:Amino Acid Sequence; D000818:Animals; D020346:Aquaporins; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D003459:Crystallins / Q000096:biosynthesis* / Q000235:genetics; D004247:DNA / Q000032:analysis; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D005136:Eye Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D018398:Homeodomain Proteins; D007381:Intermediate Filament Proteins / Q000096:biosynthesis* / Q000235:genetics; D007908:Lens, Crystalline / Q000166:cytology / Q000378:metabolism*; D008562:Membrane Glycoproteins; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D016133:Polymerase Chain Reaction; D011817:Rabbits; D012097:Repressor Proteins; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
D020346:Aquaporins; D003459:Crystallins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005136:Eye Proteins; D018398:Homeodomain Proteins; D007381:Intermediate Filament Proteins; C489025:Lim2 protein, mouse; D008562:Membrane Glycoproteins; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D012097:Repressor Proteins; D014157:Transcription Factors; C030328:aquaporin 0; C079098:filensin; C084808:phakinin; D004247:DNA