title: Cloning and characterization of a rat ortholog of MMP-23 (matrix metalloproteinase-23), a unique type of membrane-anchored matrix metalloproteinase and conditioned switching of its expression during the ovarian follicular development.
abstract: In our attempt to study the role of matrix metalloproteinases (MMPs) in the process of mammalian ovulation, we isolated a rat ortholog of the recently reported human MMP-23 from gonadotropin-primed immature rat ovaries. Transient expression of epitope-tagged rat and human MMP-23 in COS-1 cells revealed that they were synthesized as a membrane-anchored glycoprotein with type II topology. Indirect immunofluorescent analysis showed that subcellular localization of MMP-23 was predominantly in the perinuclear regions. The transfected human MMP-23 protein was processed endogenously to the soluble form in COS-1 cells. However, cotransfection of MMP-23 with the mouse furin cDNA did not enhance this processing, indicating that furin may not be involved in this event. Notably, in situ hybridization analysis revealed a dramatic switching of MMP-23 mRNA localization from granulosa cells to theca-externa/fibroblasts and ovarian surface epithelium during the follicular development. In serum-free primary culture of rat granulosa cells, a drastic diminution of MMP-23 mRNA expression was observed in response to FSH action between 24 h and 48 h of culture. The observed effect of FSH on MMP-23 expression was mimicked by treatment of granulosa cells with forskolin or 8-bromo (Br)-cAMP. In contrast, MMP-23 mRNA levels increased in theca-interstitial cells regardless of the presence of LH in the culture. However, treatment of theca-interstitial cells with forskolin or 8-Br-cAMP markedly reduced the expression of MMP-23 with a concomitant increase in progesterone production. These results indicate that the MMP-23 gene is spatially and temporally regulated in a cell type-specific manner in ovary via the cAMP signaling pathway.
authors.affiliation: Division of Biological, Graduate School of Science Hokkaido University Sapporo 060-0810, Japan. johnishi@sci.hokadai.ac.jp
authors.forename: J
authors.initials: J
authors.lastname: Ohnishi
authors.forename: E
authors.initials: E
authors.lastname: Ohnishi
authors.forename: M
authors.initials: M
authors.lastname: Jin
authors.forename: W
authors.initials: W
authors.lastname: Hirano
authors.forename: D
authors.initials: D
authors.lastname: Nakane
authors.forename: H
authors.initials: H
authors.lastname: Matsui
authors.forename: A
authors.initials: A
authors.lastname: Kimura
authors.forename: H
authors.initials: H
authors.lastname: Sawa
authors.forename: K
authors.initials: K
authors.lastname: Nakayama
authors.forename: H
authors.initials: H
authors.lastname: Shibuya
authors.forename: K
authors.initials: K
authors.lastname: Nagashima
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D005260:Female; D005640:Follicle Stimulating Hormone / Q000502:physiology; D045683:Furin; D018507:Gene Expression Regulation, Developmental; D006107:Granulosa Cells / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry; D007986:Luteinizing Hormone / Q000502:physiology; D020782:Matrix Metalloproteinases; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000201:enzymology / Q000502:physiology*; D010053:Ovary / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013381:Subtilisins / Q000494:pharmacology; D013799:Theca Cells / Q000201:enzymology
chemical_list: D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D013381:Subtilisins; D045683:Furin; C492180:MMP23A protein, human; C117705:MMP23B protein, human; D020782:Matrix Metalloproteinases; D008666:Metalloendopeptidases