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Autoradiographic detection of RNA and glycoprotein synthesis in epithelial cells of the oviduct in ovulated heifers

PIVKO J, GRAFENAU P, UHRIN V, KOPECNY V
Veterinarni Medicina 44, 1999, 41-47

Heifers (n = 9) of the Black-Pied Holstein-Friesian breed were slaughtered on the 3rd, Gth and 9th day of the synchronized cycle, respectively (heat = day 0). Tissue samples from the ampullar part of the oviduct were collected immediately after slaughter and treated for histoautoradiographic (ARG) analyses. They were cultured for 20 min on air in 0.25 ml medium Dulbecco PBS at 38 degrees C enriched with 100 mu Ci/ml of [5-H-3] uridine (specific activity 740 GBq/mM, UVVVR Prague) for detection of the RNA synthesis. For the ARG detection of glycoprotein synthesis the samples of oviductal tissue were cultured for 60 or 240 min on air, respectively, in 0.25 mi medium Dulbecco PBS at 38 degrees C to which 100 mu Ci/ml of L-[6-H-3] fucose (specific activity 0.55-1.1 TBq/mM, Amersham Int., Great Britain) was added. After incubation, the samples were fixed, dehydrated and embedded in Epon 812. Semithin (1 mu m) or ultrathin sections were coated with nuclear liquid emulsion K.5 or L.4, respectively. After one-month exposure they were developed using D 19 developer and after staining or contrasting they were observed using either light-microscopy or electron microscopy (using an electron microscope JEM 100 CX at 80 kV). Investigated autoradiograms revealed an intensive synthesis of RNA in both cilliated as well as in secretory cells of the oviduct epithelium. The intensity of RNA synthesis remains comparable during the early luteal phase from the 3rd to the 9th day of the cycle. The glycoproteins, synthesized first in the Golgi apparatus in the supranuclear region were seen to move to the apical region of the epithelial cells later on. The most intensive autoradiographic reaction of newly synthesized glycoproteins was detected on the 3rd day of the cycle without an evident change on the 6th day but decreasing on the 9th day


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