Ultrastructural localization of prolactin and chromogranin B messenger ribonucleic acids with biotinylated oligonucleotide probes in cultured pituitary cells.
A technique for the subcellular localization of prolactin and chromogranin B messenger RNAs (mRNA) in pituitary adenomas by in situ hybridization with biotinylated oligonucleotide probes is described. Ultrastructural examination revealed clusters and individual gold particles in the cytoplasm with this pre-embedding in situ hybridization method. Prolactinomas expressed both prolactin and chromogranin B mRNA, whereas the null cell adenoma expressed only chromogranin B mRNA. Sections of positively labeled cells contained up to 30 gold particles/cell. Treatment of cells with RNAse before hybridization reduced the number of gold particles to less than 1/cell. These results indicate that biotinylated oligonucleotide probes can be used to localize different mRNAs in cultured pituitary cells at the ultrastructural level to study the processing of these molecules within specific cells and for more precise correlation of molecular function with ultrastructural morphology.
Department of Pathology, University of Michigan Medical Center, Ann Arbor.
R V
RV
Lloyd
L
L
Jin
J
J
Song
D000236:Adenoma / Q000032:analysis* / Q000473:pathology; D000595:Amino Acid Sequence; D053378:Chromogranin B; D002864:Chromogranins / Q000235:genetics*; D006801:Humans; D008722:Methods; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D010911:Pituitary Neoplasms / Q000032:analysis* / Q000473:pathology; D011388:Prolactin / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D014407:Tumor Cells, Cultured
C506134:CHGB protein, human; D053378:Chromogranin B; D002864:Chromogranins; D009419:Nerve Tissue Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011388:Prolactin