Dual immunodetection and hybridisation in situ of opsin mRNA and rhodopsin protein in retinal sections.
We describe a simple method for combining in situ hybridisation and immunohistochemistry on the same retinal section. The technique was developed using a radiolabelled cDNA probe for opsin and an antibody (ROS1F4) against rhodopsin. This method retains the antigenic sites if immunocytochemistry is performed prior to in situ hybridisation. Opsin mRNA was found in the photoreceptor inner segment with rhodopsin immunolocalised to the photoreceptor outer segments. The technique should be applicable to numerous situations including analysis of the sequence of events in the expression and synthesis of the various opsins during retinal development and degeneration.
Foreman
D M
DM
University Department of Ophthalmology, Manchester Royal Eye Hospital, UK.
Jones
S
S
McKechnie
N M
NM
Williams
G
G
Boulton
M E
ME
D000911:Antibodies, Monoclonal; D015342:DNA Probes / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D012160:Retina / Q000737:chemistry*; D012243:Rhodopsin / Q000032:analysis*; D017299:Rod Opsins / Q000235:genetics*
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D012333:RNA, Messenger; D017299:Rod Opsins; D012243:Rhodopsin