Colocalization of mRNA and protein using in situ hybridization and immunohistochemistry in testicular tissue.
Using testes fixed by perfusion with Bouin's fluid and embedded in paraffin wax, this study has established methods for combining in situ hybridization and immunocytochemistry on the same section to colocalize mRNA and protein for transition protein-1 (TP-1) and sulfated glycoprotein-1 (SGP-1), respectively. It was found that SGP-1 could be detected in tissue sections subsequent to the detection of TP-1 mRNA in situ. The finding that 1) the tissue pretreatments required to permeabilize the section and to allow access to the probe, and 2) the hybridization conditions themselves, had no adverse effect on the detection of antigen, eases the performance of this technique. On this basis, important information could be obtained on the transcriptional and translational activity of spermatogenic cells, if related probes and antibodies are utilized.
Millar
M R
MR
M.R.C. Reproductive Biology Unit, Centre for Reproductive Biology, Edinburgh, U.K.
Sharpe
R M
RM
Maguire
S M
SM
Gaughan
J
J
Saunders
P T
PT
D000818:Animals; D001483:Base Sequence; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis* / Q000235:genetics; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D049231:Saposins; D013737:Testis / Q000737:chemistry*
D002868:Chromosomal Proteins, Non-Histone; D006023:Glycoproteins; C483347:Psap protein, rat; D012333:RNA, Messenger; D049231:Saposins; C053886:spermatid transition proteins