Localization of ZP3 mRNA in mouse ovary by non-radioactive in situ hybridization with digoxigenin-labelled cDNA.
In situ hybridization using digoxigenin-labelled cDNA has dual advantages of having high structural resolution and high sensitivity and avoiding problems associated with the handling of radioactive materials. We have used this technique to detect and locate mRNA for mouse ZP3 protein in paraffin sections of fixed ovaries of newborn and adult mice. Mouse ZP3 mRNA was restricted to the cytoplasm of oocytes. After day 16, ZP3 mRNA expression was increasingly restricted to the cytoplasm of the oocyte. Previous work on the distribution of ZP3 mRNA was confirmed and we were able to show that using non-isotopic in situ hybridization, sensitive and accurate cellular localization was feasible in paraffin embedded mouse ovary.
Kimura
J
J
Department of Veterinary Anatomy, College of Agriculture and Veterinary Medicine, Nihon University, Kanagawa, Japan.
Sato
K
K
Okano
M
M
Tsukise
A
A
D000469:Alkaline Phosphatase / Q000737:chemistry; D000818:Animals; D015342:DNA Probes; D004076:Digoxigenin; D004527:Egg Proteins / Q000235:genetics*; D005260:Female; D006023:Glycoproteins / Q000032:analysis; D006062:Gonadotropins / Q000494:pharmacology; D017403:In Situ Hybridization / Q000379:methods*; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000737:chemistry; D010053:Ovary / Q000737:chemistry* / Q000254:growth & development; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis*; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins
D015342:DNA Probes; D004527:Egg Proteins; D006023:Glycoproteins; D006062:Gonadotropins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse; D000469:Alkaline Phosphatase; D004076:Digoxigenin