Hormonal regulation of pituitary FSH sialylation in male rats

Ambao, Verónica; Rulli, Susana B.; Carino, Mónica Herminia; Cónsole, Gloria M.; Ulloa‐Aguirre, Alfredo; Calandra, Ricardo S.; Campo, Stella

doi:10.1016/j.mce.2009.05.002

Cited by 12 publications

(18 citation statements)

References 46 publications

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“…In contrast, mammalian cells are able to express glycoproteins with complex sialylated oligosaccharides [42]. Functionally, GTH isoforms containing more sialylated oligosaccharides are less biopotent, as has been shown for rat FSH [41] and LH [40]. This information supports our findings that Sf9-Fsh is more bioactive than CHO-Fsh.…”

Section: Discussionsupporting

confidence: 90%

“…When we analyzed the potency and B:I ratio, Sf9-Fsh was more bioactive than CHO-Fsh but oddly less than CHO-scFsh. These different bioactivities of the recombinant GTHs could be due to a different degree and type of glycosylation according to the expression system used [39][40][41]. Insect cells produce glycoproteins containing high mannose-type oligosaccharides.…”

Section: Discussionmentioning

confidence: 99%

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Receptor Specificity and Functional Comparison of Recombinant Sea Bass (Dicentrarchus labrax) Gonadotropins (Fsh and Lh) Produced in Different Host Systems1

Molés

Zanuy

Muñoz

et al. 2011

Biology of Reproduction

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Different yields, biopotency, and in vivo pharmacokinetics are obtained for recombinant sea bass gonadoltropins depending on the production system and DNA construct, but they show specific activation of their corresponding receptors. Gonadotropins (GTHs) are glycoprotein hormones that play a major role in the regulation of gonadal functions. Recently, we succeeded in isolating the native sea bass Fsh from sea bass pituitaries, but to ensure the availability of bioactive GTHs and no cross-contamination with other related glycoproteins, recombinant sea bass GTHs were produced using two expression systems-insect and mammalian cells-and different constructs that yielded tethered or noncovalently bound dimers. Their production levels, binding specificity to their homologous cognate receptors, and bioactivity were investigated and compared. Both expression systems were successful in the generation of bioactive recombinant GTHs, but insect Sf9 cells yielded higher amounts of recombinant proteins than mammalian Chinese Hamster Ovary (CHO) stable clones. All recombinant GTHs activated their cognate receptors without cross-ligand binding and were able to stimulate sea bass gonadal steroidogenesis in vitro, although with different biopotencies. To assess their use for in vivo applications, their half-life in sea bass plasma was evaluated. Sf9-GTHs had a lower in vivo stability compared with CHO-GTHs due to their rapid clearance from the blood circulation. Cell-dependent glycosylation could be contributing to the final in vivo stability and biopotency of these recombinant glycoproteins. In conclusion, both insect and mammalian expression systems produced bioactive sea bass recombinant gonadotropins, although with particular features useful for different proposes (e.g., antibody production or in vivo studies, respectively).

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Receptor Specificity and Functional Comparison of Recombinant Sea Bass (Dicentrarchus labrax) Gonadotropins (Fsh and Lh) Produced in Different Host Systems1

Molés

Zanuy

Muñoz

et al. 2011

Biology of Reproduction

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show abstract

“…The data obtained showed that the insect-derived Sf9-Fsh was more biopotent than CHO-Fsh, but less so than the single-chain CHO-scFsh (Molés et al, 2011a). These differences could be due to a different type and degree of glycosylation depending on the expression system used (Ambao et al, 2009;Grossmann et al, 1997;Olivares et al, 2009). It is known that insect cells, unlike mammalian cells, are not able to add sialic acid residues, producing instead glycoproteins that contain high mannose-type oligosaccharides (Kost et al, 2005).…”

Section: Biopotency Of the Recombinant Gonadotropinsmentioning

confidence: 98%

“…The results showed that CHO-produced single-chain gonadotropins are more stable in plasma than Sf9-derived gonadotropins (Molés et al, 2011a). This behavior could be due to glycosylation differences, since the content in terminal sialic acid residues determines the rate at which glycoproteins are cleared from circulation (Ambao et al, 2009;Olivares et al, 2009;Ulloa-Aguirre et al, 2001). Moreover, the fusion of the two subunits and the presence of O-linked oligosaccharides in CTP may additionally contribute to prolonging the circulating half-life of single-chain gonadotropins (Ben-Menahem and Boime, 1996;Fares et al, 1992;Klein et al, 2003).…”

Section: In Vivo Stabilitymentioning

confidence: 99%

“…More specific studies in mammals have demonstrated that changes in the content of sialic acid affect the bioactivity of Fsh isoforms. In humans and rats, Fsh variants with more acidic/sialylated glycosylation exhibit a longer plasma half-life but lower receptor binding activity and in vitro biological potency than their less acidic counterparts (Ambao et al, 2009;Zambrano et al, 1996). In addition, in mammals, it has been observed that the molecular microheterogeneity of intrapituitary FSH may change depending on the age, sexual development, and/or the steroidogenic milieu, provoking a range of biological responses (Ambao et al, 2009;Rulli et al, 1999).…”

Section: Tools To Measure Gonadotropin Subunitsmentioning

confidence: 99%

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Gonadotropins in European sea bass: Endocrine roles and biotechnological applications

Mazón

Molés

Rocha

et al. 2015

General and Comparative Endocrinology

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Synthesis and secretion of gonadotropins including structure-function correlates

Bousfield

Dias

2011

Rev Endocr Metab Disord

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The synthesis and secretion of the gonadotropic hormones involves coordination of signal transduction, gene expression, protein translation, post-translational folding and modification and finally secretion. The production of biologically active gonadotropin thus requires appropriately folded and glycosylated subunits that assemble to form the heterodimeric hormone. Here we overview recent literature on regulation of gonadotropin subunit gene expression and current understanding of the assembly and secretion of biologically active gonadotropic hormones. Finally, we discuss the therapeutic potential of understanding glycosylation function towards designing new forms of gonadotropins based on observations of physiologically relevant parameters such as age related glycosylation changes.

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Hormonal regulation of pituitary FSH sialylation in male rats

Cited by 12 publications

References 46 publications

Receptor Specificity and Functional Comparison of Recombinant Sea Bass (Dicentrarchus labrax) Gonadotropins (Fsh and Lh) Produced in Different Host Systems1

Receptor Specificity and Functional Comparison of Recombinant Sea Bass (Dicentrarchus labrax) Gonadotropins (Fsh and Lh) Produced in Different Host Systems1

Gonadotropins in European sea bass: Endocrine roles and biotechnological applications

Synthesis and secretion of gonadotropins including structure-function correlates

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