Expression of functional nerve growth factor receptors in pancreatic beta-cell lines and fetal rat islets in primary culture

Diabetes. 1993 Dec;42(12):1829-36. doi: 10.2337/diab.42.12.1829.

Abstract

Previous data demonstrated that one rat insulinoma cell line, RINm5F cells, which is a rat beta-cell line derived from a pancreatic tumor, express mRNA coding for both the low- and the high-affinity nerve growth factor receptors. Goals of this study were to extend our data to other beta-cell lines and fetal islets in primary culture and to study further the binding characteristics of nerve growth factor receptors on beta-cells. Northern blot analysis revealed that not only a panel of endocrine beta-cell lines (RINm5F, INS-1, beta-TC3) but also fetal rat islets in primary culture express mRNA coding for trk-A, which has been proposed to be the neuronal high-affinity nerve growth factor receptors. Reverse polymerase chain reaction followed by sequencing revealed that the sequence of trk-A receptor in RINm5F cells is identical to that of trk-A expressed in PC12 cells. The expression of the low-affinity nerve growth factor receptor was examined by Northern blot analysis that showed low-affinity nerve growth factor receptor to be expressed in RINm5F and INS-1 cell lines, in fetal rat islets in primary culture, but not in beta-TC3-cells. Binding experiments revealed the presence of low- and high-affinity nerve growth factor binding sites, identical to those described for PC12 cells, on RINm5F and INS-1 cells and only high-affinity binding sites on beta-TC3 cells. Exposure of all three beta-cell lines to nerve growth factor increased NGFI-A and c-fos mRNA steady-state levels, showing that these receptors are functional.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cells, Cultured
  • DNA Primers
  • Fetus
  • Gene Expression*
  • Genes, fos
  • Insulinoma
  • Islets of Langerhans / metabolism*
  • Kinetics
  • Molecular Sequence Data
  • Nerve Growth Factors / metabolism
  • PC12 Cells
  • Pancreatic Neoplasms
  • Polymerase Chain Reaction
  • Proto-Oncogene Proteins / biosynthesis*
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-fos / biosynthesis
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / metabolism*
  • Rats
  • Rats, Wistar
  • Receptor Protein-Tyrosine Kinases / biosynthesis*
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Receptor, trkA
  • Receptors, Nerve Growth Factor / biosynthesis*
  • Receptors, Nerve Growth Factor / metabolism

Substances

  • DNA Primers
  • Nerve Growth Factors
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-fos
  • RNA, Messenger
  • Receptors, Nerve Growth Factor
  • Receptor Protein-Tyrosine Kinases
  • Receptor, trkA