Effects of basic fibroblast growth factor and nerve growth factor on lactate production, gamma-glutamyl transpeptidase and aromatase activities in cultured Sertoli cells

Schteingart, Helena Fedora; Meroni, Silvina Beatriz; Canepa, DF; Pellizzari, Eliana Herminia; Cigorraga, SB

doi:10.1530/eje.0.1410539

Cited by 25 publications

(19 citation statements)

References 40 publications

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“…Sertoli cells from 20-day-old Sprague-Dawley rats were isolated as previously described (Meroni et al 1999). Briefly, decapsulated testes were digested with 0·1% collagenase and 0·006% soybean trypsin inhibitor in Hanks' balanced salt solution for 5 min at room temperature.…”

Section: Sertoli Cell Isolation and Culturementioning

confidence: 99%

“…Germ cells are a potential source of this peptide although other cells in the testis also express it (Mullaney & Skinner 1992, Han et al 1993. bFGF receptors are constitutively expressed in Sertoli cell cultures (Le Magueresse-Battistoni et al 1994) and it has been shown that bFGF modulates, among other responses, the number of follicle-stimulating hormone (FSH) receptors (Jaillard et al 1987), transferrin, estradiol and lactate secretion (Han et al 1993, Schteingart et al 1999, glucose uptake, lactate dehydrogenase (LDH) activity and the levels of glucose transporter 1 and LDH A mRNA .…”

Section: Introductionmentioning

confidence: 99%

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Assessment of the roles of mitogen-activated protein kinase and phosphatidyl inositol 3-kinase/protein kinase B pathways in the basic fibroblast growth factor regulation of Sertoli cell function

Riera

Meroni²,

Pellizzari³

et al. 2003

Journal of Molecular Endocrinology

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Basic fibroblast growth factor (bFGF) belongs to the large set of intratesticular regulators that provide the fine tuning of cellular processes implicated in the maintenance of spermatogenesis. The aim of the present study was to determine the participation of mitogen-activated protein kinase (MAPK) and phosphatidyl inositol 3-kinase/protein kinase B (PI3K/PKB) pathways in bFGF regulation of Sertoli cell function. Twenty-day-old rat Sertoli cell cultures were used. Stimulation of the cultures with bFGF showed a time-dependent increment in phosphorylated MAPK and PKB levels that reached maximal values in 5-min incubations. MAPK kinase inhibitors U0126 (U) and PD98059 (PD) and a PI3K inhibitor wortmannin (W) were able to block the stimulatory effects of bFGF on phosphorylated MAPK and PKB levels respectively. The participation of MAPK-and PI3K/PKB-signaling pathways in the regulation by bFGF of two well-known Sertoli cell-differentiated functions, lactate and transferrin production, was next explored. As for lactate production, PD and W did not modify the ability of bFGF to stimulate lactate production. However, a combination of PD and W partially impaired the increase in lactate production elicited by bFGF. The participation of MAPK-and PI3K/PKB-signaling pathways in the regulation by bFGF of glucose uptake and lactate dehydrogenase (LDH) activity was also analysed. In this respect, it was observed that W markedly decreased basal and bFGF-stimulated glucose uptake and that U and PD did not modify it. On the other hand, U and PD decreased the stimulation of LDH activity by bFGF whereas W did not modify it. As for transferrin production, while both MAPK kinase inhibitors partially decreased the ability of bFGF to stimulate transferrin secretion, the PI3K inhibitor did not modify it. In summary, the results demonstrated that bFGF stimulates MAPK-and PI3K/PKB-dependent pathways in rat Sertoli cells. Moreover, these results showed that while bFGF utilizes the MAPK pathway to regulate transferrin production and LDH activity, it uses the PI3K/PKB pathway to regulate glucose transport into the cell.

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Section: Sertoli Cell Isolation and Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Assessment of the roles of mitogen-activated protein kinase and phosphatidyl inositol 3-kinase/protein kinase B pathways in the basic fibroblast growth factor regulation of Sertoli cell function

Riera

Meroni²,

Pellizzari³

et al. 2003

Journal of Molecular Endocrinology

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“…Figure 1 shows the dose-response curve to bFGF (0·1-50 ng/ml) for rat Sertoli cell lactate production. Significant increments in lactate production at doses equal to or higher than 10 ng/ml were observed as previously described (Schteingart et al 1999). To ascertain the possible effect of residual contaminating germ cells on bFGF regulation of lactate production, Sertoli cell monolayers were hypotonically treated (HT) and compared with untreated controls in their ability to produce lactate.…”

Section: Discussionmentioning

confidence: 73%

“…While the functional role of bFGF in the testis is still under study, it is known to have several modulatory effects in Sertoli cell cultures, where bFGF receptors are constitutively expressed (Le Magueresse-Battistoni et al 1994). Here, the growth factor modulates DNA synthesis (Jaillard et al 1987), release of transferrin (Han et al 1993), estradiol production, -glutamyl transpeptidase activity and lactate secretion (Schteingart et al 1999). Additionally, bFGF modulates expression of FSH receptors (Smith et al 1989), plasminogen activator inhibitor-1 (Le MagueresseBattistoni et al 1998), cfos, jun B (Smith et al 1989) and Sertolin (Mruk & Cheng 1999).…”

Section: Introductionmentioning

confidence: 99%

“…As for the LDH isoenzyme system, increments in lactate production in Sertoli cells have been correlated with an increase in the LDH5 isoenzyme -containing four subunits A. Glucose transport through the plasma membrane and LDH A mRNA levels are regulated in a distinct manner by FSH (Riera et al 2001), interleukin 1 (IL1) (Nehar et al 1998), tumor necrosis factor-(TNF ) (Nehar et al 1997) and epidermal growth factor (EGF) (Boussouar & Benahmed 1999) -factors that modify Sertoli cell lactate production. No data are available on the mechanisms involved in the recently described effect of bFGF on lactate production (Schteingart et al 1999).…”

Section: Introductionmentioning

confidence: 99%

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Regulation of lactate production and glucose transport as well as of glucose transporter 1 and lactate dehydrogenase A mRNA levels by basic fibroblast growth factor in rat Sertoli cells

Riera

Meroni²,

Schteingart³

et al. 2002

Journal of Endocrinology

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By using cultured rat Sertoli cells as a model, both the action of basic fibroblast growth factor (bFGF) on lactate production and the site of this action were studied. bFGF stimulated Sertoli cell lactate production in a dosedependent manner (basal: 7·3 0·5; 0·1 ng/ml bFGF: 7·5 0·5; 1 ng/ml bFGF: 7·5 0·6; 10 ng/ml bFGF: 10·3 1·0; 30 ng/ml bFGF: 15·2 1·5; 50 ng/ml bFGF: 15·4 1·6 µg/µg DNA). Two major sites for the action of this growth factor were identified. First, bFGF was shown to exert short-and long-term stimulatory effects on glucose transport (basal: 1170 102; 30 ng/ml bFGF for 120 min: 1718 152 and basal: 718 64; 30 ng/ml bFGF for 48 h: 1069 69 d.p.m./µg DNA respectively). Short-term bFGF stimulation of glucose transport was not inhibited by the protein synthesis inhibitor cycloheximide. These results indicate that short-term bFGF stimulation of glucose uptake does not involve an increase in the number of glucose transporters. On the other hand, stimulation with bFGF for periods of time longer than 12 h increased glucose transporter 1 (GLUT1) mRNA levels. These increased mRNA levels were probably ultimately responsible for the increments in glucose uptake that are observed in long-term treated cultures. Secondly, bFGF increased lactate dehydrogenase (LDH) activity (basal: 31·0 1·4; 30 ng/ml bFGF: 45·7 2·4 mIU/µg DNA). The principal subunit component of those LDH isozymes that favors the transformation of pyruvate to lactate is subunit A. bFGF increased LDH A mRNA levels in a dose-and time-dependent manner. In summary, the results presented herein show that glucose transport, LDH activity and GLUT1 and LDH A mRNA levels are regulated by bFGF to achieve an increase in lactate production. These observed regulatory actions provide unequivocal evidence of the participation of bFGF in Sertoli cell lactate production which may be related to normal germ cell development.

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β‐nerve growth factor participates in an auto/paracrine pathway of regulation of the meiotic differentiation of rat spermatocytes

Perrard

Vigier

Damestoy

et al. 2006

Journal Cellular Physiology

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NGF appears to be involved in spermatogenesis. However, mice lacking NGF or TrkA genes do not survive more than a few days whereas p75 NTR knockout mice are viable and fertile. Therefore, we addressed the effect of bNGF on spermatogenesis by using the systems of rat germ cell culture we established previously. bNGF did not modify the number of Sertoli cells, pachytene spermatocytes, secondary spermatocytes nor the half-life of round spermatids, but increased the number of secondary meiotic metaphases and decreased the number of round spermatids formed in vitro. These effects of bNGF were reversible and maximal at about 4 Â 10 À11 M. Conversely, K252a, a Trk-specific kinase inhibitor, enhanced the number of round spermatids above that of control cultures. The presence of bNGF and its receptors TrkA and p75 NTR was investigated in testis sections, in Sertoli cell and germ cell fractions, and in germ cell and Sertoli cell co-cultures. bNGF was detected only in germ cells from pachytene spermatocytes of stages VII up to spermatids of stages IX-X. TrkA and p75 NTR were detected in Sertoli cells and in these germ cells. Taken together, these results indicate that bNGF should participate in an auto/paracrine pathway of regulation of the second meiotic division of rat spermatocytes in vivo.

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Effects of basic fibroblast growth factor and nerve growth factor on lactate production, gamma-glutamyl transpeptidase and aromatase activities in cultured Sertoli cells

Cited by 25 publications

References 40 publications

Assessment of the roles of mitogen-activated protein kinase and phosphatidyl inositol 3-kinase/protein kinase B pathways in the basic fibroblast growth factor regulation of Sertoli cell function

Assessment of the roles of mitogen-activated protein kinase and phosphatidyl inositol 3-kinase/protein kinase B pathways in the basic fibroblast growth factor regulation of Sertoli cell function

Regulation of lactate production and glucose transport as well as of glucose transporter 1 and lactate dehydrogenase A mRNA levels by basic fibroblast growth factor in rat Sertoli cells

β‐nerve growth factor participates in an auto/paracrine pathway of regulation of the meiotic differentiation of rat spermatocytes

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