Rat testicular N-cadherin: its complementary deoxyribonucleic acid cloning and regulation

Endocrinology. 1998 Apr;139(4):1853-62. doi: 10.1210/endo.139.4.5958.

Abstract

Using primer sets specific for mouse N-cadherin and rat testicular RNA for RT-PCR, a full-length complementary DNA (cDNA) coding for rat testicular N-cadherin was isolated. The deduced amino acid sequence of rat N-cadherin yielded a 883-amino acid polypeptide that displayed a 98.6% identity with the mouse homolog. N-Cadherin was found to be expressed by Sertoli and germ cells in the rat testis by RT-PCR. Using Sertoli-germ cell cocultures, it was found that the N-cadherin expression increased with time in culture. To assess whether this is due to a soluble factor(s) released from germ cells that affects Sertoli cell N-cadherin expression, germ cell-conditioned media (GCCM) were fractionated by preparative anion-exchange HPLC, and the resulting fractions were divided into 14 pools. Pool 4 was found to contain a factor(s) that induced a dose-dependent stimulation on Sertoli cell N-cadherin expression with a maximal stimulation at 2 microg protein/dish/4.5 x 10(6) Sertoli cells. At higher doses between 12 and 32 microg protein/dish, this pool relinquished its effect on Sertoli cell N-cadherin expression suggestive of a biphasic effect. This biphasic effect was confirmed using increasing doses of crude GCCM on Sertoli cell cultures. Since nonviable germ cells failed to stimulate Sertoli cell N-cadherin expression, it illustrates the observed stimulatory effect by GCCM is likely to be mediated via a soluble factor(s) releasing from viable germ cells. These results reveal the presence of a stimulatory factor(s) in GCCM that can modulate Sertoli cell N-cadherin expression in vitro. Since N-cadherin plays a crucial role in facilitating invasive capacity of metastatic tumor cells, the observation of germ cell-released factor(s) in affecting Sertoli cell N-cadherin expression may suggest its possible role in facilitating germ cell migration during spermatogenesis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cadherins / chemistry
  • Cadherins / genetics*
  • Cells, Cultured
  • Cloning, Molecular*
  • Culture Media, Conditioned
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics*
  • Gene Expression
  • Male
  • Mice
  • Molecular Sequence Data
  • RNA, Messenger / analysis
  • Rats
  • Rats, Sprague-Dawley
  • Sequence Homology
  • Sertoli Cells / chemistry
  • Spermatozoa / chemistry
  • Spermatozoa / metabolism
  • Testis / chemistry*

Substances

  • Cadherins
  • Culture Media, Conditioned
  • DNA, Complementary
  • RNA, Messenger

Associated data

  • GENBANK/AF097593