Differential effects of follicle-stimulating hormone glycoforms on the transcriptome profile of cultured rat granulosa cells as disclosed by RNA-seq
Zariñán, Teresa Espinal-Enriquez, Jesús De Anda-Jáuregui, Guillermo Lira-Albarrán, Saúl Hernández-Montes, Georgina Gutiérrez-Sagal, Rubén Rebollar-Vega, Rosa G. Bousfield, George R. Butnev, Viktor Y. Hernández-Lemus, Enrique
Zariñán, Teresa
Espinal-Enriquez, Jesús
De Anda-Jáuregui, Guillermo
Lira-Albarrán, Saúl
Hernández-Montes, Georgina
Gutiérrez-Sagal, Rubén
Rebollar-Vega, Rosa G.
Bousfield, George R.
Butnev, Viktor Y.
Hernández-Lemus, Enrique
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2024
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Follitropin,Transcriptome,Animal,Cell culture,CHO cell line,Cricetulus,Drug effect,Female,Glycosylation,Granulosa cell,Human,Metabolism,Procedures,Rat,RNA sequencing,Animals,Cells,Cultured,CHO Cells,Cricetulus,Female,Follicle Stimulating Hormone,Glycosylation,Granulosa Cells,Humans,Rats,RNA-Seq,Transcriptome
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Zariñán, T., Espinal-Enriquez, J., De Anda-Jáuregui, G., Lira-Albarrán, S., Hernández-Montes, G., Gutiérrez-Sagal, R., Rebollar-Vega, R.G., Bousfield, G.R., Butnev, V.Y., Hernández-Lemus, E., Ulloa-Aguirre, A. Differential effects of follicle-stimulating hormone glycoforms on the transcriptome profile of cultured rat granulosa cells as disclosed by RNA-seq. (2024). PLoS ONE, 19 (6.0), art. no. e0293688. DOI: 10.1371/journal.pone.0293688
Abstract
It has been documented that variations in glycosylation on glycoprotein hormones, confer distinctly different biological features to the corresponding glycoforms when multiple in vitro biochemical readings are analyzed. We here applied next generation RNA sequencing to explore changes in the transcriptome of rat granulosa cells exposed for 0, 6, and 12 h to 100 ng/ml of four highly purified follicle-stimulating hormone (FSH) glycoforms, each exhibiting different glycosylation patterns: a. human pituitary FSH$^{18/21}$ (hypo-glycosylated); b. human pituitary FSH$^{24}$(fully glycosylated); c. Equine FSH (eqFSH) (hypo-glycosylated); and d. Chinese-hamster ovary cell-derived human recombinant FSH (recFSH) (fully-glycosylated). Total RNA from triplicate incubations was prepared from FSH glycoform-exposed cultured granulosa cells obtained from DES-pretreated immature female rats, and RNA libraries were sequenced in a HighSeq 2500 sequencer (2 x 125 bp paired-end format, 10-15 x 10$^6$ reads/sample). The computational workflow focused on investigating differences among the four FSH glycoforms at three levels: gene expression, enriched biological processes, and perturbed pathways. Among the top 200 differentially expressed genes, only 4 (0.6%) were shared by all 4 glycoforms at 6 h, whereas 118 genes (40%) were shared at 12 h. Follicle-stimulating hormone glycocoforms stimulated different patterns of exclusive and associated up regulated biological processes in a glycoform and time-dependent fashion with more shared biological processes after 12 h of exposure and fewer treatment-specific ones, except for recFSH, which exhibited stronger responses with more specifically associated processes at this time. Similar results were found for down-regulated processes, with a greater number of processes at 6 h or 12 h, depending on the particular glycoform. In general, there were fewer downregulated than upregulated processes at both 6 h and 12 h, with FSH$^{18/21}$ exhibiting the largest number of down-regulated associated processes at 6 h while eqFSH exhibited the greatest number at 12 h. Signaling cascades, largely linked to cAMP-PKA, MAPK, and PI3/AKT pathways were detected as differentially activated by the glycoforms, with each glycoform exhibiting its own molecular signature. These data extend previous observations demonstrating glycosylation-dependent distinctly different regulation of gene expression and intracellular signaling pathways triggered by FSH in granulosa cells. The results also suggest the importance of individual FSH glycoform glycosylation for the conformation of the ligand-receptor complex and induced signalling pathways. © 2024 Zariñán et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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©2024 Zariñán et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Public Library of Science
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PLoS ONE
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1932-6203