Neural cell adhesion molecules in rat endocrine tissues and tumor cells: distribution and molecular analysis.
The adhesive properties of neural cell adhesion molecules (NCAMs) can be modified by alternative splicing of the primary transcript or posttranslational modifications. In the present study, we describe distinct forms of alternative splicing and posttranslational modification of the extracellular domain of NCAM of various endocrine tissues and derived tumor cells of the rat. Using an antiserum detecting the immunoglobulin-like domains of NCAM as well as a monoclonal antibody recognizing the NCAM-specific polysialic acid (PSA), we observed a similar staining pattern in adrenals, pituitary, and neoplastic endocrine cells. In endocrine tumor cells [pheochromocytoma (PC12), insulinoma (RINA2), and pituitary tumor cells (GH3)], NCAM immunoreactivity was most intense at contact sites between the cells. The immunocytochemical data were substantiated by results of in situ hybridization histochemistry. Specifically, higher levels of NCAM mRNA were detected in the adrenal cortex than in the medulla. In the pituitary, NCAM mRNA was more abundant in the anterior and intermediate lobes than in the neural lobe. The sequence of NCAM mRNAs in endocrine cells was analyzed by polymerase chain reaction and S1 nuclease protection assays. We found that major exons 4-13 of the NCAM mRNA in endocrine tissues and related tumor cell lines were homologous to those in the brain. However, PC12, RINA2, and GH3 tumor cells; normal rat pituitaries; and adrenals contained different amounts of NCAM mRNA with an alternative extra exon, termed VASE (also called pi in mouse) between constitutive exons 7 and 8. In addition, in pituitaries, we detected an alternative exon in splice site a between the constitutive exons 12 and 13, termed a15, with or without an AAG triplett. These sites are thought to be important for the adhesive properties of NCAM. Therefore, these results suggest that modifications of NCAM may be important for adhesive interactions in normal and neoplastic endocrine cells.
Lahr
G
G
Abteilung Anatomie und Zellbiologie, Universität Ulm, Germany.
Mayerhofer
A
A
Bucher
S
S
Barthels
D
D
Wille
W
W
Gratzl
M
M
D000311:Adrenal Glands / Q000502:physiology*; D017398:Alternative Splicing; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015816:Cell Adhesion Molecules, Neuronal / Q000032:analysis* / Q000235:genetics*; D002460:Cell Line; D015342:DNA Probes; D005091:Exons; D005260:Female; D015870:Gene Expression; D014644:Genetic Variation; D007106:Immune Sera; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016716:PC12 Cells; D010902:Pituitary Gland / Q000502:physiology*; D010911:Pituitary Neoplasms; D016133:Polymerase Chain Reaction; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015816:Cell Adhesion Molecules, Neuronal; D015342:DNA Probes; D007106:Immune Sera; D009838:Oligodeoxyribonucleotides; D015347:RNA Probes; D012333:RNA, Messenger