2005 Zoological Society of Japan
ZOOLOGICAL SCIENCE 22: 237–245 (2005)
Changes in the Distribution of Tenascin and Fibronectin in the Mouse Ovary During Folliculogenesis, Atresia, Corpus Luteum Formation and Luteolysis Keiko Yasuda*, Emi Hagiwara, Akiko Takeuchi, Chinatsu Mukai†, Chiyuki Matsui‡, Atsushi Sakai and Satoshi Tamotsu Department of Biological Science, Faculty of Science, Nara Women’s University, Nara 630-8506, Japan
ABSTRACT—Tenascin and fibronectin are components of the extracellular matrices that oppose and promote adhesion, respectively. Using immunohistochemical techniques, we studied the distribution of tenascin and fibronectin in the mouse ovary, in which dynamic reconstruction and degeneration occur during folliculogenesis, atresia, ovulation, corpus luteum formation and luteolysis. In growing follicles, tenascin was only detected in the theca externa layer, while fibronectin was detected in the theca externa layer, theca interna layer and basement membrane. During follicular atresia, granulosa cells, which are surrounded by the basement membrane, began to die through apoptosis. In atretic follicles, tenascin was detected in the basement membrane and theca externa layer. Distribution of fibronectin in atretic follicles was similar to that in healthy growing follicles, except that granulosa cells were slightly immunopositive for fibronectin. In young corpus luteum, luteal cells exhibit high 3 β -hydroxysteroid dehydrogenase (3 β -HSD) activity, an enzyme indispensable for progesterone production. Tenascin was barely detected in young luteal cells. 3 β -HSD activity in luteal cells declines with corpus luteum age, and in older corpus luteum there is an increase in apoptotic death of luteal cells. Tenascin was intensely immunopositive in old luteal cells.In contrast, fibronectin immunostaining in luteal cells was relatively constant during corpus luteum formation and luteolysis. Our observations suggest that tenascin is critical in controlling the degenerative changes of tissues in mouse ovaries. Moreover, in all circumstances observed in this study, tenascin always co-localized with fibronectin, suggesting fibronectin is indispensable for the function of tenascin. Key words: tenascin, fibronectin, atresia, luteolysis, ovary
INTRODUCTION Extracellular matrices play various important roles in controlling cell behavior and function. They provide mechanical strength to the tissue, promote or inhibit cell adhesion, and guide movement of cells. Moreover, extracellular matrices bind various growth factors, thereby modulating their local concentrations and actions in tissue (Vigny et al., 1988; Vukicevic et al., 1992; Asem et al., 2000). Tenascin is a large, oligomeric extracellular matrix pro* Corresponding author. Phone: +81-742-20-3412; Fax : +81-742-20-3412; E-mail:
[email protected] † Present address: Department of Life Science, Graduate School of Art and Science, University of Tokyo, Komaba 153-8902, Japan ‡ Present address: KAN Research Institute, Science Center Build. Kyoto Research Park, 1 Chudoji-Awatacho, Shimogyo-ku, Kyoto 600-8815, Japan
tein with a six-armed hexa-brachion structure (Erickson et al., 1989; Chiquet, 1989; Spring et al, 1989). It shows both temporally and spatially restricted tissue distribution during development in kidneys (Aufderheide et al., 1987), nerves (Wehrle et al., 1990) and the cranial neural crest (BronnerFraser et al., 1988). Tenascin is also observed in tissues where regeneration and reconstruction occur, for example, in a healing skin wound (Mackie et al, 1988) and around a regenerating nerve (Danilof et al., 1989). Tenascin is thought to counteract adhesion and allow tissue to move apart by increasing the secretion of protease from cells (Werb et al., 1990) and resulting in disruption of the extracellular matrix network. Fibronectin is another kind of extracellular matrix molecule, the structure of which is similar to tenascin (Spring et al., 1989). One function of fibronectin is to serve as a general adhesive molecule, linking cells one another or to other substrates such as collagen and proteoglycans.
238
K. Yasuda et al.
Attachment of tenascin and fibronectin to cells is mediated by multiple integrins (Prieto et al., 1993; Schnapp et al., 1995; Koivisto et al., 2000; Sriramarao et al., 1993; Loike et al., 2001). Several integrins that mediate tenascin function can also recognize fibronectin (Schnapp et al., 1995; Doane et al., 2002). Moreover, tenascin can interfere with the function of fibronectin (Chiquet-Ehrismann et al., 1988; Pesheva et al., 1994; Deryugina et al., 1996; Probstmeier et al., 1999). When tenascin and fibronectin are simultaneously present in the same tissue, therefore, their relative amounts might regulate the strength of cell attachment and, consequently, cell functions. In ovaries, primordial follicles periodically start to develop; some of them ovulate and form corpora lutea, while others fail to ovulate and undergo atresia. Since reconstruction and degeneration always take place in ovaries, antiadhesion molecules, such as tenascin, might play important roles in regulating ovarian functions. A marked increase in the tenascin immunostaining was observed in the intercellular space of regressing corpora lutea during late luteal phase in human ovaries (Tamura et al., 1993), suggesting the involvement of tenascin in luteolysis. However, changes in the distribution of tenascin during other phases of an ovarian cycle (i.e., folliculogenesis, atresia, corpus luteum formation and luteolysis) have not been examined in detail. Moreover, we know quite little about the relationship between fibronectin and tenascin in the regulation of ovarian function, although there are some reports on the roles of fibronectin alone in ovarian functions (Aten et al., 1995; Huet et al., 1997, 1998, 2001). In the present study, we examined the distributions of tenascin and fibronectin in the mouse ovary during folliculogenesis, atresia, corpus luteum formation and luteolysis by immunohistochemistry. Our results suggest different roles for these two extracellular matrix proteins in the regulation of ovarian function.
MATERIALS AND METHODS Chemicals and Reagents Pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (hCG), and anti-mouse fibronectin polyclonal antibody were obtained from Biogenesis Ltd. (England, UK). Anti-mouse tenascin monoclonal antibody and bovine serum albumin (BSA) were from Sigma Chemical Co. (MO, USA). Dehydroepiandrostenedione was from Nacalai Tesque Inc. (Kyoto, Japan). Nicotinamide adenine dinucleotide, tetranitro blue tetrazolium and diaminobenzidine (DAB) were from Wako Pure Chemical Industries Ltd. (Osaka, Japan). Animals and hormonal treatment Female ICR mice were kept under controlled light condition (14h light, 10h dark), and given food and water ad libitum. Our investigations were conducted in accordance with the Guide for Care and Use of Laboratory Animals. Immature mice (3 weeks old) were injected subcutaneously (sc) with 5 IU of PMSG, a treatment to initiate and synchronize the development of follicles up to the preovulatory stages. At 48 hours after PMSG injection, the mice were injected sc with 5 IU of hCG.
Ovulation occurred about 10 hours after hCG treatment. The hormones used in the present study, PMSG and hCG, have similar functions to follicle-stimulating hormone (FSH) and luteinizing hormone (LH), respectively. Immunohistochemistry Unfixed ovaries were embedded in OCT compound (Sakura Finetechnical Co. Tokyo Japan) and frozen on dry ice immediately after collection. Cryostat sections were cut at 7 µm and stained with antibodies to mouse tenascin or fibronectin using the ABC method (Vectastain ABC system, Vector laboratories, CA USA). In short, sections were rinsed with PBS and incubated in 0.3% hydrogen peroxide in methanol for 30 min to block endogenous peroxidases. After washing in PBS, the sections were subsequently incubated in normal serum for 30 min to reduce non-specific staining, and incubated overnight with anti-mouse tenascin antibody at dilution of 1/ 500,000 or anti-mouse fibronectin antibody at dilution of 1/100,000 in 0.5% BSA/PBS, respectively, in a humidified chamber at room temperature. After a rinse in PBS, the sections were incubated for 30 min with biotinylated secondary antibody, rinsed again in PBS and incubated for 60 min with an avidin-biotin-peroxidase complex. The sections were washed again in PBS, and peroxidase activity was visualized with 0.05% DAB in PBS containing 0.03% hydrogen peroxide for 7 min. Finally, the sections were rinsed in distilled water, dehydrated and mounted. Immunohistochemical localization of tenascin or fibronectin was observed by differential-interferencecontrast microscopy (Leitz DMR). Negative control sections to assess non-specific staining were incubated in 0.5% BSA/PBS instead of primary antibody. No staining was evident in the absence of primary antibody. 3 β -Hydroxysteroid dehydrogenase (3 β -HSD) staining To detect 3 β -HSD activity in an ovary, the cryostat sections of ovaries were air dried, and incubated for 30 min at 37°C in the reaction mixture, which was composed of dehydroepiandrostenedione, nicotinamide adenine dinucleotide, and tetranitro blue tetrazolium, as described by Wattenberg (1958). Nuclei of the cells were then counter-stained with methyl green. Apoptosis detection Apoptosis in ovarian cells was detected by Tunel staining using the ApopTag In Situ Detection Kit (Intergen company, NY USA). In brief, frozen sections were fixed with 4% paraformaldehyde in PBS (pH 7.4) and incubated in 3.0% hydrogen peroxide for 5 min to quench endogenous peroxidase. Digoxygenin labeled nucleotides were added to the free 3'OH DNA termini by terminal deoxynucleotidyl transferase for 1 hour at 37°C. The sections were then incubated with anti-digoxigenin antibody conjugated to a peroxidase reporter protein at room temperature for 30 min. After wash in PBS, peroxidase activity was visualized with DAB.
RESULTS Growing follicles During folliculogenesis, tenascin and fibronectin exhibited different localization patterns. As shown in Fig. 1, in a preantral follicle, the oocyte is surrounded by several layers of granulosa cells, which are further surrounded by a basement membrane (a thin mat composed of several extracellular matrices), two or three layers of theca interna cells and one or two layers of theca externa cells, in this order. Tenascin was detected in the theca externa layer (Fig. 1-a), but was not detected in the theca interna or basement membrane. In contrast, fibronectin was detected not only in theca
Tenascin and Fibronectin in Ovary
239
Fig. 1. Immunolocalization of tenascin (a) and fibronectin (b) in a growing mouse follicle. a), b) show neighboring sections of the same follicle. Arrows indicate the primary follicle that contains no theca layer. Arrowheads indicate the position of the follicular basement membrane. BM, basement membrane; G, granulosa cell layers; O, oocyte; TI, theca interna; TE, theca externa. Bar=100 µm.
externa but also in theca interna and basement membrane (Fig. 1-b). In immature follicles in which theca cell layers have not yet formed (indicated by arrows in Fig. 1-a, 1-b), tenascin and fibronectin were detected on the borderline between follicles and ovarian interstitial cells surrounding them. The distribution of tenascin and fibronectin in the follicles at later developmental stages (antral and preovulatory follicles) was similar to that in the preantral follicles (data not shown). Although the interstitial cell surface was usually immunostained with both anti-tenascin and anti-fibronectin antibodies, the intensities of immunostaining observed there were relatively low. Oocytes and granulosa cells was not immunostained with antibodies to either of the extracellular matrices throughout folliculogenesis (Fig. 1-a and 1-b).
Fig. 2. Localization of apoptotic cells (a), tenascin (b) and fibronectin (c) in an atretic mouse follicle. a), b) and c) show neighboring sections of the same follicle. Note the localization of tenascin to the basement membrane in atretic follicle. Oocyte is not included in b). BM: basement membrane; G, granulosa cell layer; O, oocyte; TE, theca externa; TI, theca interna. Bar=100 µm
240
K. Yasuda et al.
Atretic follicles Follicles that fail to ovulate enter the atretic pathway. During this process, the theca interna cells increase in size and become cuboid (hypertrophy). Within the basement membrane the granulosa cell layers become irregular and apoptosis of cells in the granulosa cell layer is observed (Fig. 2-a). In atretic follicles, tenascin was detected not only in the theca externa (as in the healthy growing follicles shown in Fig. 1-a) but also in the basement membrane (Fig. 2-b). In contrast, the theca interna, granulosa cells and the oocyte showed no signal for tenascin (Oocyte was not shown in Fig. 2-b). In the atretic follicles, fibronectin was detected in the theca externa, theca interna, and basement membrane, as in the growing follicles, although the signal intensity in the basement membrane was weak and discontinuous (Fig. 2-c), as compared with that of healthy growing
follicles (Fig. 1-b). In the atretic follicles, fibronectin was also detected in the intercellular spaces of the granulosa cells but was not detected in oocytes. Ovulation and corpus luteum formation Ten hours after hCG injection, follicles begin ovulating, and subsequently corpus luteum formation begins. Before ovulation, granulosa cell layers within the basement membrane are not vascularized with capillaries. During corpus luteum formation, the basement membrane breaks down and capillary sprouts arising from venules in the theca interna invade the cavity of the ruptured follicle. The granulosa cells and theca interna cells undergo luteinization, while the theca externa, which represents the outermost boundary of the corpus luteum, remains unchanged. During the process of luteinization, granulosa cells and theca interna cells
Fig. 3. Detection of 3 β -hydroxysteroid dehydrogenase (a), tenascin (b, d), and fibronectin (c, e) in follicles immediately after ovulation. b) c), and d) e) show neighboring sections in the same follicle, respectively. Arrows indicate the sprout of venule from theca interna cell layer. Bars=100 µm.
Tenascin and Fibronectin in Ovary
become hypertrophic and begin to secrete a great deal of progesterone. Three β -HSD is the enzyme that catalyses conversion of pregnenolone to progesterone and increases markedly at luteinization. Therefore, luteal cells were identified on the basis of steroidogenic function, using 3 β -HSD activity as a marker. Fig. 3 shows the distribution patterns of 3 β -HSD activity (Fig. 3-a), tenascin (Fig. 3-b and 3-d) and fibronectin (Fig.
241
3-c and Fig. 3-e) in the mouse ovary 14 hours after hCG injection (4 hours after ovulation). Judging from the distribution of 3 β -HSD within the follicle (Fig. 3-a), luteinization progressed from the periphery towards the center of the follicle. Capillary sprouts invading the cavity of ruptured follicle were present within the 3 β -HSD positive area (observed as negative images in Fig. 3-a), but had not yet reached the central region of the ruptured follicle, where 3 β -HSD activ-
Fig. 4. Detection of apoptosis (a, e, i), 3 β -hydroxysteroid dehydrogenase activity (b, f, j), tenascin (c, g, k) and fibronectin (d, h, l) in corpora lutea one day (a–d), two days (e–h), and three days (i–l) after hCG injection. Four photographs in the same day show neighboring sections of the same corpus luteum. Each corpus luteum is bordered on the right by a follicle in order to compare the signal intensities between luteal cells (CL) and granulosa cell layer (G) in the same photograph. Arrows in c) and g) show the capillaries in the corpus luteum. CL, corpus luteum; G, granulosa cell layer of follicle; Tc, theca externa of corpus luteum; Tf, theca cell layers of follicle. Bar=100 µm.
242
K. Yasuda et al.
ity was still low. These results suggest a close relationship between capillary extension and luteal differentiation (high 3 β -HSD activity). Observations at higher magnification (Fig. 3-b and Fig. 3-c) revealed that the newly formed sprout was always accompanied by tenascin and fibronectin, suggesting that both tenascin and fibronectin are involved in capillary extension in the initial phase of corpus luteum formation. The tenascin (Fig. 3-d) and fibronectin (Fig. 3-e) positive region subsequently expanded from the periphery towards the central region of the ruptured follicle, in conjunction with capillary sprout extension. Tenascin positive areas coincided with fibronectin positive areas and areas of capillary extension (compare Fig. 3-d and Fig. 3-e). Moreover, tenascin immunostaining was especially strong around the capillary (Fig. 3-d; Capillaries are indicated by arrows in Fig. 3-d and Fig. 3-e), but relatively weak in newly luteinized cells around them, while fibronectin immunostaining was of uniform intensity both around the capillaries and in newly luteinized cells (Fig. 3-e). The detection of tenascin in newly luteinized cells rapidly disappeared, while fibronectin was able to be detected for a longer period. Corpus luteum and luteolysis We then examined alterations in tenascin and fibronectin localization at various stages of corpora lutea maturation (Fig. 4; For comparison, parts of neighboring corpora lutea and follicles are shown in Fig4-a to 4-i). One day after hCG injection (Day1; that is, 14 hours after ovulation), the corpus luteum had already formed. The newly formed corpus luteum contained no apoptotic cells (Fig. 4-a) and showed high 3 β -HSD activity (Fig. 4-b). A few apoptotic cells appeared in corpus luteum on Day 2 (Fig. 4-e), when 3 β -HSD activity of luteal cells still remained high (Fig. 4-f). The number of apoptotic cells increased on Day 3 (Fig. 4-i), in association with an abrupt decline in 3 β -HSD activity (Fig. 4-j). These results indicate that regression of corpus luteum begins 3 days after hCG injection. On Day1 (Fig. 4-c) and Day2 (Fig. 4-g) post hCG injection, luteal cells weakly stained with anti-tenascin antibody or were negative for tenascin, but immunostaining signals for tenascin were exceptionally strong around the capillary (indicated by arrows in Fig. 4-c and 4-g). Although tenascin immunostaining was observed in newly luteinized cells immediately after ovulation (Fig. 3-d), it was not again observed in luteal cells until Day 3 (at a start of luteolysis), at which point tenascin staining in luteal cells became intense (Fig. 4-k). The theca externa, the outermost boundary of corpus luteum, exhibited relatively constant and strong signals for tenascin throughout the process from corpus luteum formation to luteolysis (Fig. 4-c, g and k). Fibronectin was present in the luteal cells and theca externa on Day1 (Fig. 4-d), and its staining intensity in luteal cells did not change much thereafter (Fig. 4-d, h and l). Thus, the ratio of tenascin to fibronectin appeared to change during luteal regression. These results suggest a correlation between increased amounts of tenascin and
regression of luteal cells. DISCUSSION Hormones and local regulators, such as growth factors and cytokines, have been considered to control ovarian function. In addition, extracellular matrices are also candidates as important regulators of ovarian function, as they might regulate proliferation and differentiation of granulosa cells (Amsterdam et al., 1989; Richardson et al., 1992; Luck, 1994; Aharoni et al., 1996; Sites et al., 1996). In the present study, we demonstrate that tenascin shows stage-specific localization to distinct tissues in the mouse ovary, suggesting its importance in the regulation of ovarian functions. In contrast, fibronectin distribution is wider and more constant than tenascin, suggesting its role as a rather basic extracellular matrix. In follicles While fibronectin is thought to be an adhesive molecule (Rouslahti and Pierschbacher, 1986), tenascin seems to be an anti-adhesive molecule (Chiquet-Ehrismann et al., 1988; Spring et al., 1989). In growing follicles, the theca externa layer, which represents the outermost surface of follicles, was observed to contain both tenascin and fibronectin, whereas theca interna and basement membrane expressed fibronectin only (Fig. 1). Theca cells attach to each other to enclose granulosa cells and an oocyte, suggesting that fibronectin should play a central role in the architecture of theca cell layers and maintenance of follicle integrity during folliculogenesis. On the other hand, tenascin in the theca externa might give a follicle the ability to migrate within an ovary, by detaching from the interstitial cells around it. The ontogeny of theca cells is still obscure. Although theca cells seemed to be derived from ovarian interstitial cells (Hirshfield, 1991; Hisaw, 1947; Byskov, 1980; Guraya, 1985; Erickson et al., 1985), experimental evidence to support this hypothesis is lacking. Theca cells form two layers (theca interna layer and theca externa layer), but the process of the differentiation of the two theca cell layers is also unclear. At luteinization, theca interna cells are transformed into luteal cells, while theca externa cells are not. During folliculogenesis, however, the distinction between the theca interna and theca externa becomes less clear. Markers to distinguish theca interna cells from theca externa cells are indispensable to investigate the differentiation of theca cells, yet there have been few reports on such cellular markers. Our study reveals that theca externa layers contain both tenascin and fibronectin, whereas theca interna layers contain only fibronectin. Such differences in the distribution of tenascin and fibronectin were consistently observed from young follicles to preovulatory follicle. Therefore, absence of tenascin may be a marker to distinguish theca interna cells from theca externa cells. Tenascin was detected as a component of the basement membrane of an atretic follicle, whereas it was not
Tenascin and Fibronectin in Ovary
detected in the basement membrane of a growing follicle. Although presence of tenascin in the ‘follicular wall’ (a region containing granulosa cell layers, basement membrane, and theca cell layers) of human atretic follicle has been reported (Tamura et al., 1993), the localization of tenascin at the cellular level has not been defined. Thus, this is the first report of the presence of tenascin in the basement membrane of the atretic follicle. The basement membranes in growing follicles are thought to support proliferation of granulosa cells and prevent the cells from undergoing apoptosis (Aharoni et al., 1996). The role of the basement membrane in suppression of programmed cell death has also been reported in mammary epithelium (Pullan et al., 1996). In atretic follicles, granulosa cells detach from the basement membranes, stop proliferating, and show apoptotic features, as tenascin appears in the basement membrane. Because tenascin is thought to exhibit anti-adhesion activity by inducing the secretion of metalloproteinase from the cells in contact with it (Danilof et al., 1989), it is suggested that tenascin in the basement membrane of atretic follicles might accelerate the disruption of the attachment of granulosa cells to the basement membrane, resulting in the enhanced apoptotic degeneration of granulosa cell layers. In contrast to tenascin, fibronectin shows rather constant immunolocalization in the basement membrane of both atretric follicles and growing follicles. It is, therefore, suggested that fibronectin might not be related to the atretic pathway in the mouse ovary. There are some conflicting data on the role of fibronectin in induction and/or repression of apoptosis. Sugahara et al. (1994) suggested that fibronectin could induce apoptosis in human hematopoietic cell lines, whereas Zhang et al. (1995) demonstrated that fibronectin supported cell survival in Chinese hamster ovary cell line. However, in rat granulosa cells isolated from preovulatory follicles, fibronectin failed to protect granulosa cells from apoptosis (Aharoni et al., 1996). Thus, the effects of fibronectin on apoptosis remain unclear at present. In corpus luteum Ovulation is a drastic process accompanied by tissue re-organization. Just after ovulation, remaining granulosa and theca interna cells differentiate into luteal cells. The first step of the luteinization process begins with break down of the basement membrane. Capillaries from the theca interna layer invade the cavity of the ruptured follicle. In bovine corpus luteum, fibronectin was detected in early capillary sprouts from venules in the theca interna layer (Amselgruber et al., 1999). We found that not only fibronectin but also tenascin around the capillary sprouts in the mouse luteinizing follicle. Considering the proposed role for tenascin in opposing adhesion, tenascin might be involved in cell movement during capillary sprout formation and extension. During corpus luteum formation, the changes in tenascin localization differed from that of fibronectin localization. Although distribution of tenascin (Fig. 3-d) and fibronectin (Fig. 3-e) expanded from the periphery towards the central
243
region during corpus luteum formation, only fibronection was present in luteal cells of the newly established corpus luteum (Fig. 4-c, 4-d). In early and mature luteal cells (1 day or 2 days after hCG injection), intense 3 β -HSD activity was detected, but tenascin was hardly detected except for around capillaries. Three days after hCG injection, when the corpus luteum began to regress and 3 β -HSD activity abruptly declined, there was a marked increase in tenascin expression in luteal cells. Our results are in good agreement with data from the human corpus luteum, which show a more intense signal for tenascin in the late luteal phase as compared with the midluteal phase (Tamura et al., 1993). Our results suggest that tenascin might not be necessary for 3 β -HSD activity of luteal cells; it may instead play a role in repressing progesterone production in regressing corpus luteum. In contrast with tenascin, fibronectin was continuously and widely present in luteal cells of mouse ovaries from corpus luteum formation till luteolysis. In bovine corpora lutea, fibronectin was also observed from early luteal phase to late luteal phase. (Zhao and Luck, 1995; Silvester and Luck, 1999; De Candia and Rodgers, 1999). Fibronectin, therefore, might play an important basic role in corpus luteum function but not in its formation or regression. Relationship between tenascin and fibronectin In our results, tenascin always appeared in association with fibronectin, while fibronectin could be present without tenascin. The co-distribution of tenascin with fibronectin has also been reported in cultured fibroblasts (Uysal and Hemming, 1999) and in the amphibian embryo (Riou et al., 1988). Tenascin has anti-adhesive properties for various cell types attaching to fibronectin (Chiquet-Ehrismann et al., 1988; Pesheva et al., 1994; Deryugina et al., 1996; Probstmeier and Pesheva, 1999). In rabbit synovial fibroblasts, tenascin induced the secretion of collagenase (a metalloproteinase), only when fibronectin was simultaneously present (Tremble et al., 1994). These results suggest that tenascin might require the presence of fibronectin to exert its antiadhesive effects. However, the relation between tenascin and fibronectin in ovary is still obscure. What kind of factors regulates the expression of tenascin and fibronectin in the ovary? In the mouse ovary, tenascin showed the stage-specific, spatially distinct distribution in relation to ovarian cycle stage, and although fibronectin showed relatively wide and constant distribution, its distribution also changed at luteinization. The hormonal milieu in an ovary drastically changes around luteinization. Potential candidates for regulators of fibronection and tenascin are, therefore, gonadotropins (especially luteinizing hormone), and steroids (such as progesterone and 17 β -estradiol). In chicken granulosa cells, luteinizing hormone stimulates fibronectin production via progesterone (Asem and Conkright, 1995). On the other hand, local regulators, such as growth factors and cytokines, are other candidates. Rattig et al. (1994) reported that tenascin synthesis in fibroblasts
244
K. Yasuda et al.
was induced by inteleukin-1, tumor necrosis factor- α and interleukin-4. In mouse ovaries, distribution pattern of tenascin was different from that of fibronectin, suggesting that the synthesis of tenascin and fibronectin was regulated by different factors in the mouse ovary. Ovarian function is controlled by a network of local regulators as well as endocrine regulators, and the present data reveal that the extracellular matrix molecules, tenascin and fibronectin, are important members of this network. We are currently investigating the interactions between tenascin and fibronectin in the mouse ovary using an in vitro luteinized granulosa cell culture system (manuscript in preparation). However, means by which tenascin and fibronectin associate with other regulators to influence ovarian functions must be further investigated.
REFERENCES Aharoni D, Meiri I, Atzmon R, Vlodavsky I, Amsterdam A (1996) Differential effect of components of the extracellular matrix on differentiation and apoptosis. Curr Biol 7: 43–51 Amselgruber WM, Schafer M, Sinowatz F (1999) Angiogenesis in the bovine corpus luteum: An Immunocytochemistry and ultrastructural study. Anat Histol Embryol 28: 157–166 Amsterdam A, Rotmensch S, Furman A, Venter EA, Vlodavsky I (1989) Synergistic effect of human chorionic gonadotropin and extracellular matrix on in vitro differentiation of human granulosa cells: progesterone production and gap junction formation. Endocrinology 124: 1956–1964 Asem EK, Conkright MD (1995) Role of progesterone in luteinizing hormone-induced fibronectin production and deposition by chicken granulosa cells in vitro. Comp Biochem Physiol 112c: 247–255 Asem EK, Stingley-Salazar SR, Robinson J.P, Turek JJ (2000) Identification of some components of basal lamina of avian ovarian follicle. Poult Sci 79: 589–601 Aten RF, Kolodecik TR, Behrman HR (1995) A cell adhesion receptor antiserum abolishes, whereas laminin and fibronectin glycoprotein components of extracellular matrix promote, luteinization of cultured rat granulosa cells. Endocrinology 136: 1753–1758 Aufderheide E, Chiquet-Ehrismann R, Ekblom P (1987) Epithelialmesenchymal interactions in the developing kidney lead to expression of tenascin in the mesenchyme. J Cell Biol 105: 599–608 Byskov AG (1980) Sexual differentiation of the mammalian ovary. In “Biology of the Ovary” Ed by PM Motta, ESE Hafez, Martinus Nijhoff, The Hague, pp 3–15 Bronner-Fraser M (1988) Distribution and function of tenascin during cranial neural crest development in the chick. J Neurosci Res 21: 135–147 Chiquet M (1989) Tenascin/J1/cytotactin: the potential function of hexabrachion proteins in neural development. Dev Neurosci 11: 266–275 Chiquet-Ehrismann R, Kalla P, Pearson CA, Beck K, Chiquet M (1988) Tenascin interferes with fibronectin action. Cell 53: 383390 Danilof JK, Crossin KL, Pinçon-Raymond M, Murawsky M, Rieger F, Edelman GM (1989) Expression of cytotactin in the normal and regenerating neuromuscular system. J Cell Biol 108: 625–635 De Candia LM, Rodgers RJ (1999) Characterization of the expression of the alternative splicing of the ED-A, ED-B and V regions of fibronectin mRNA in bovine ovarian follicles and corpora
lutea. Reprod Fertil Dev 11: 367–377 Deryugina EI, Bourdon MA (1996) Tenascin mediates human glioma cell migration and modulates cell migration on fibronectin. J Cell Sci 109: 643–652 Doane KJ, Bhattacharya R, Marchant J (2002) Perturbation of 1 integrin function using anti-sense or function-blocking antibodies on corneal cells grown on fibronectin and tenascin. Cell Biol Int 26: 131–144 Erickson, GF, Magoffin DA, Dyer CA, Hofeditz C (1985) The ovarian androgen producing cells: a review of structure/function relationships. Endocr Rev 6: 371–399 Erickson HP, Bourdon, MP (1989) Tenascin: an extracellular matrix protein prominent in specialized embryonic tissues and tumors. Annu Rev Cell Biol 5: 71–92 Guraya SS (1985) Biology of ovarian follicles. Springer-Verlag, New York, pp 79–85 Hirshfield AN (1991) Theca cells may be present at the outset of follicular growth. Biol Reprod 44: 1157–1162 Hisaw FL (1947) Development of the Graafian follicle and ovulation. Physiol Rev 27: 95–119 Huet C, Monget P, Pisselet C, Monniaux D (1997) Changes in extracellular matrix components and steroidogenic enzymes during growth and atresia of antral ovarian follicles in the sheep. Biol Reprod 56: 1025–1034 Huet C, Monget P, Pisselet C, Hennequet C, Locatelli A, Monniaux D (1998) Chronology of events accompanying follicular atresia in hypophysectomized ewes. Changes in levels of steroidogenic enzymes, connexin 43, insulin-like growth factor II/ mannose 6 phosphate receptor, extracellular matrix components, and matrix metalloproteinases. Biol Reprod 58: 175–185 Huet C, Pisselet C, Mandon-Pepin B, Monget P, Monniaux D (2001) Extracellular matrix regulates ovine granulosa cell survival, proliferation and steroidogenesis: relationships between cell shape and function. J Endocrinol 169: 347–360 Koivisto L, Grenman R, Heino J, Larjava H (2000) Integrins α5β1, αVβ1, and αVβ6 collaborate in squamous carcinoma cell spreading and migration on fibronectin. Exp Cell Res 255: 10– 17 Loike JD, Cao L, Budhu S, Hoffman S, Silverstein SC (2001) Blockage of α5β1 integrins reverses the inhibitory effect of tenascin on chemotaxis of human monocytes and polymorphonuclear leukocytes through three-dimensional gels of extracellular matrix proteins. J Immunol 166: 7534–7542 Luck MR (1994) The gonadal extracellular matrix. Oxf Rev Reprod Biol 16: 33–85 Mackie EJ, Halfter W, Liverani D (1988) Induction of tenascin in healing wounds. J Cell Biol 107: 2757–2767 Pesheva P, Probstmeier R, Skubitz APN, McCarthy JB, Furcht LK, Schachner M (1994) Tenascin-R (J1 160/180) inhibits fibronectin-mediated cell adhesion-functional relatedness to tenascinC. J Cell Sci 107: 2323–2333 Prieto AL, Edelman GM, Crossin KL (1993) Multiple integrins mediate cell attachement to cytotactin/tenascin. Proc Natl Acad Sci USA 90: 10154–10158 Probstmeier R, Pesheva P (1999) Tenascin-C inhibits β1 integrindependent cell adhesion and neurite outgrowth on fibronectin by a disialoganglioside-mediated signaling mechanism. Glycobiology 9: 101–114 Pullan S, Wilson J, Metcalfe A, Edwards GM, Goberdhan N, Tilly J, Hickman JA, Dive C, Streuli CH (1996) Requirement of basement membrane for the suppression of programmed cell death in mammary epithlium. J Cell Sci 109: 631–642 Rettig WJ, Erickson HP, Albino AP, Garin-Chesa P (1994) Induction of human tenascin (neuronectin) by growth factors and cytokines: cell type-specific signals and signaling pathways. J Cell Sci 107: 487– 497 Richardson MC, Davies DW, Watson RH, Dunsford ML, Inman CB,
Tenascin and Fibronectin in Ovary Masson GM (1992) Cultured human granulosa cells as a model for corpus luteum function: relative roles of gonadotrophin and low density lipoprotein studied under defined culture conditions. Hum Reprod 7: 12–18 Riou JF, Shi DL, Chiquet M, Boucant JC (1988) Expression of tenascin in response to neural induction in amphibian embryos. Development 104: 511–524 Ruoslahti E, Pierschbacher MD (1986) Arg-gly-asp: A versatile cell recognition signal. Cell 44: 517–518 Schnapp LM, Hatch N, Ramos DM, Klimanskaya IV, Sheppard D, Pytela R (1995) The human integrin α8β1 functions as a receptor for tenascin, fibronectin, and viteronectin. J Biol Chem 270: 23196–23202 Silvester LM, Luck MR (1999) Distribution of extracellular matrix components in the developing ruminant corpus luteum: a wound repair hypothesis for luteinization. J Reprod Fertil 116:187–198 Sites CK, Kessel B, LaBarbera R (1996) Adhesion proteins increase cellular attachment, follicle-stimulating hormone receptors, and progesterone production in cultured porcine granulosa cells. Proc Soc Exp Biol Med 212: 78–83 Spring J, Beck K, Chiquet-Ehrismann R (1989) Two contrary functions of tenascin: Dissection of the active sites by recombinant tenascin fragments. Cell 59: 325–334 Sriramarao P, Mendler M, Bourdon MA (1993) Endotherial cell attachment and spreading on human tenascin is mediated by α2β1 and αVβ3 integrins. J Cell Sci 105: 1001–1012 Sugahara H, Kanakura Y, Furitsu T, Ishihara K, Oritani K, Ikeda H, Kitayama H, Ishikawa J, Hashimoto K, Kanayama Y, Matsuzawa Y (1994) Induction of programmed cell death in human hematopoietic cell lines by fibronectin via its interaction with very late antigen 5. J Exp Med 179: 1757–1766 Tamura M, Sasano H, Suzuki T, Fukaya T, Watanabe T, Kusakabe M, Sakakura T, Yajima A (1993) Distribution of tenascin in normal cycling human ovary. Hum Reprod 8: 2015–2018
245
Tremble P, Chiquet-Ehrismann R, Werb Z (1994) The extracellular matrix ligands fibronectin and tenascin collaborate in regulating collagenase gene expression in fibroblasts. Mol Biol Cell 5: 439– 453 Uysal H, Hemming FW (1999) Changes in the expression and distribution of fibronectin, laminin and tenascin by cultured fibroblasts from skin lesion of patients with tuberous sclerosis. Br J Dermatol 141: 658–666 Vigny M, Ollier-Hartmann MP, Lavigne M, Fayein N, Jeanny JC, Laurent M, Courtois Y (1988) Specific binding of basic fibroblast growth factor to basement membrane-like structures and to purified heparan sulfate proteoglycan of the EHS tumor. J Cell Physiol 137: 321–328 Vukicevic S, Kleinman HK, Luyten FP, Roberts AB, Roche NS, Reddi AH (1992) Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components. Exp Cell Res 202: 1–8 Wattenberg LW (1958) Microscopic histochemical demonstration of steroid-3- β ol dehydrogenase in tissue sections. J Histochem Cytochem 6: 225–232 Wehrle B, Chiquet M (1990) Tenascin is accumulated along developing peripheral nerves and allows neurite outgrowth in vitro. Development 110: 401– 415 Werb Z, Tremble P, Damsky CH (1990) Regulation of extracellular matrix degradation by cell-extracellular matrix interactions. Cell Differ Dev 32: 299–306 Zhang Z, Vouri K, Reed JC, Ruoslahti E (1995) The α5β1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression. Proc Natl Acad Sci USA 92: 6161–6165 Zao Y, Luck MR (1995) Gene expression and protein distribution of collagen, fibronectin and laminin in bovine follicles and corpora lutea. J Reprod Fertil 104: 115–123 (Received August 24, 2004 / Accepted November 16, 2004)
