Lactate Dehydrogenase and Malate Dehydrogenase of Sertoli Cells in Rats

Santiemma, V.; Salfi, V.; Casasanta, N.; Fabbrini, A

doi:10.3109/01485018708986800

Cited by 18 publications

(7 citation statements)

References 23 publications

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“…After sequential enzymatic digestion with 0.25% trypsin and 0.1% collagenase (Boehringer Mannheim, Mannheim, Germany) as previously described [17], purified peritubular myoid cells were obtained by a Percoll discontinuous gradient technique (Tung and Fritz [2]). Briefly, serial dilutions of Percoll (Pharmacia, Uppsala, Sweden) from 70% to 30% were prepared.…”

Section: Tpmc Isolation and Culturementioning

confidence: 99%

Endothelin-1 Stimulates Deoxyribonucleic Acid Synthesis and Contraction in Testicular Peritubular Myoid Cells1

Santiemma¹,

Beligotti²,

Magnanti³

et al. 1996

Biology of Reproduction

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In the testis, endothelin-1 (ET-1) is produced by Sertoli cells, and it has been proposed to be a paracrine factor participating in the regulation of tubular and interstitial function. The response of purified testicular peritubular myoid cells (TPMC) to ET-1 was investigated in the present study. TPMC expressed a single class of high-affinity receptors that were shown by competitive binding experiments with sarafotoxin-6c to belong to the ETA subtype. The binding of ET-1 to TPMC was followed by rapid internalization of the receptor-ligand complex. ET-1 induced a prompt rise in intracellular Ca2+ concentration that was blunted in Ca(2+)-free medium and in the presence of Mn2+ or of voltage-operated-calcium-channel (VOC) blockers, indicating that both Ca2+ mobilization from intracellular stores and extracellular Ca2+ influx were involved. Thymidine uptake was promoted by ET-1 in a time-dependent manner, and the use of cyclo[D-Asp-L-Pro-D-Val-L-Leu-D-Trp] (BQ123) reduced the incorporation of thymidine. Protein kinase C (PKC) inhibition (100 nM calphostin C) abolished the ET-1 mitogenic effect. ET-1 also promoted TPMC contraction, as evaluated in collagen lattices, in a dose-related manner, with the half-maximal response observed at 1 nM. As in the case of mitogenesis, BQ123 blunted ET-1-induced contraction. PKC inhibition abolished ET-1-induced contraction. These findings indicate that ET-1 promotes DNA synthesis and contraction of TPMC and that both effects are mediated by PKC; they suggest as well that ET-1 may have a physiological role in the interaction between Sertoli cells and TPMC.

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Section: Tpmc Isolation and Culturementioning

confidence: 99%

Endothelin-1 Stimulates Deoxyribonucleic Acid Synthesis and Contraction in Testicular Peritubular Myoid Cells1

Santiemma¹,

Beligotti²,

Magnanti³

et al. 1996

Biology of Reproduction

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“…LDH is a well characterized isozyme that is involved in the inter‐conversion of pyruvate into lactate (Santiemma et al ., ; Li, ). Hormone‐induced augmentation of LDH activity in 18d Sc as compared with that in 60d Sc may explain the reason of failure of FSH to augment lactate production by 60d Sc.…”

Section: Discussionmentioning

confidence: 99%

Hormone responsiveness of cultured Sertoli cells obtained from adult rats after their rapid isolation under less harsh conditions

et al. 2016

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SUMMARYDuring adulthood, testicular Sertoli cells (Sc) coordinate all stages of germ cell (Gc) development involved in sperm production. However, our understanding about the functions of adult Sc is limited because of the difficulties involved in the process of isolating these cells from the adult testis, mainly because of the presence of large number of advanced Gc which interfere with Sc isolation at this age. Most of our knowledge about Sc function are derived from studies which used pre-pubertal rat Sc (18 AE 2-day old) as it is easy to isolate and culture Sc at this age. To this end, we established a less time consuming and less harsh procedure of isolating Sc from adult (60 days of age) rat testis for facilitating research on Sc-mediated regulation of spermatogenesis during adulthood. The cells were isolated using collagenase digestion at higher temperature, reducing the exposure time of cells to the enzyme.Step-wise digestion with intermittent removal of small clusters of tissue helped in increasing the yield of Sc. Isolated Sc were cultured and treated with FSH and testosterone (T) to evaluate their hormone responsiveness in terms of lactate, E 2 , cAMP production. Adult Sc were found to be active and produced high amounts of lactate in a FSH-independent manner. FSH-mediated augmentation of cAMP and E 2 production by adult Sc was less as compared with that by pre-pubertal Sc obtained from 18-day-old rats. Androgen-binding ability of adult Sc was significantly higher than pre-pubertal Sc. Although T treatment remarkably augmented expression of Claudin 11, it failed to augment lactate production by adult Sc. This efficient and rapid procedure for isolation and culture of functionally viable adult rat Sertoli cells may pave the way for determining their role in regulation and maintenance of spermatogenesis.

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“…The latter gives rise to LDH C4 isozyme present only in the mature testis, specifically in spermatozoa (Markert et al 1975). The combinations of the other two gene products result in five tetrameric LDH isozymes, LDH-1 (B4), LDH-2 (A1B3), LDH-3 (A2B2), LDH-4 (A3B1) and LDH-5 (A4) that are present in variable proportions in different somatic tissues including rat Sertoli cells (Santiemma et al 1987). LDH-1 (heart) has a lower K m for pyruvate but it is inhibited by high concentrations -like those existing in Sertoli cells -of this substrate.…”

Section: Discussionmentioning

confidence: 99%

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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Lactate Dehydrogenase and Malate Dehydrogenase of Sertoli Cells in Rats

Cited by 18 publications

References 23 publications

Endothelin-1 Stimulates Deoxyribonucleic Acid Synthesis and Contraction in Testicular Peritubular Myoid Cells1

Endothelin-1 Stimulates Deoxyribonucleic Acid Synthesis and Contraction in Testicular Peritubular Myoid Cells1

Hormone responsiveness of cultured Sertoli cells obtained from adult rats after their rapid isolation under less harsh conditions

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

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