title: Rat testicular N-cadherin: its complementary deoxyribonucleic acid cloning and regulation.
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.
authors.affiliation: Population Council, Center for Biomedical Research, New York, New York 10021, USA.
authors.forename: S S
authors.initials: SS
authors.lastname: Chung
authors.forename: M Y
authors.initials: MY
authors.lastname: Mo
authors.forename: B
authors.initials: B
authors.lastname: Silvestrini
authors.forename: W M
authors.initials: WM
authors.lastname: Lee
authors.forename: C Y
authors.initials: CY
authors.lastname: Cheng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015820:Cadherins / Q000737:chemistry / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017385:Sequence Homology; D012708:Sertoli Cells / Q000737:chemistry; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism; D013737:Testis / Q000737:chemistry*
chemical_list: D015820:Cadherins; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D012333:RNA, Messenger