Differential gene expression within the ovine corpus luteum: identification of secreted protein acidic and rich in cysteine as a major secretory product of small but not large luteal cells.
Limited information is available regarding secretory proteins of the corpus luteum (CL), and the potential local role these proteins may play in control of luteal function. An ovine small luteal cell complementary DNA library was immunologically screened with a polyclonal antiserum generated against small cell secretory proteins. A relatively abundant complementary DNA (approximately 2.1 kb) encoding a calcium binding glycoprotein Secreted Protein Acidic and Rich in Cysteine (SPARC) was isolated. Production of SPARC protein by ovine luteal cells was confirmed by immunoprecipitating it from labeled culture medium. Expression of SPARC messenger RNA (mRNA) was examined within CL collected on days 3, 7, 10, 13, and 16 post estrus (n = 4, 4, 4, 3, and 4 respectively), and within pools of purified small (n = 3) and large (n = 4) luteal cells by Northern and dot blot analysis. Amounts of SPARC mRNA increased during the early luteal phase, peaked by day 7 (P < 0.05) and subsequently declined on days 10 and 13 (P < 0.05). SPARC mRNA content was significantly higher in the small than in the large cells (P < 0.003). In situ hybridization showed that SPARC mRNA was localized to the thecal layer of Graafian follicles and to day 3 and day 10 CL. Within CL, immunohistochemistry indicated that SPARC protein was associated with small luteal cells (spindle shaped, avg = 17 microM in diameter) but not with large cells. This specific localization to small cells was confirmed by colocalization of SPARC with 3 beta-hydroxysteroid dehydrogenase. We conclude that SPARC is a major secretory product of small steroidogenic luteal cells of the ovine CL. As SPARC is known to modulate many aspects of tissue reorganization, expression by small luteal cells may play a role in regulation of CL maturation.
Smith
G W
GW
Department of Animal Sciences, University of Missouri, Columbia 65211, USA.
Gentry
P C
PC
Long
D K
DK
Smith
M F
MF
Roberts
R M
RM
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism* / Q000502:physiology*; D005260:Female; D015870:Gene Expression; D007150:Immunohistochemistry; D008183:Luteal Phase; D015676:Osteonectin / Q000235:genetics* / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012756:Sheep / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution
D015676:Osteonectin; D012333:RNA, Messenger; D015096:3-Hydroxysteroid Dehydrogenases