Rat testicular N-cadherin: its complementary deoxyribonucleic acid cloning and regulation.
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.
Chung
S S
SS
Population Council, Center for Biomedical Research, New York, New York 10021, USA.
Mo
M Y
MY
Silvestrini
B
B
Lee
W M
WM
Cheng
C Y
CY
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*
D015820:Cadherins; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D012333:RNA, Messenger