Abstract

Because of the deep involvement of granulosa cells in the processes surrounding the cycles of menstruation and reproduction, there is a great need for a deeper understanding of the ways in which they function during the various stages of those cycles. One of the main ways in which the granulosa cells influence the numerous sex associated processes is hormonal interaction. Expression of steroid sex hormones influences a range of both primary and secondary sexual characteristics, as well as regulate the processes of oogenesis, folliculogenesis, ovulation, and pregnancy. Understanding of the exact molecular mechanisms underlying those processes could not only provide us with deep insight into the regulation of the reproductive cycle, but also create new clinical advantages in detection and treatment of various diseases associated with sex hormone abnormalities. We have used the microarray approach validated by RT-qPCR, to analyze the patterns of gene expression in primary cultures of human granulosa cells at days 1, 7, 15, and 30 of said cultures. We have especially focused on genes belonging to ontology groups associated with steroid biosynthesis and metabolism, namely “Regulation of steroid biosynthesis process” and “Regulation of steroid metabolic process”. Eleven genes have been chosen, as they exhibited major change under a culture condition. Out of those, ten genes, namely STAR, SCAP, POR, SREBF1, GFI1, SEC14L2, STARD4, INSIG1, DHCR7, and IL1B, belong to both groups. Patterns of expression of those genes were analyzed, along with brief description of their functions. That analysis helped us achieve a better understanding of the exact molecular processes underlying steroid biosynthesis and metabolism in human granulosa cells.

Highlights

  • The increase of knowledge about the processes underlining the development of human gametes have brought into light the unique interactions between the tissues involved in that process

  • The genes we aim to identify might later serve for markers for the processes, both normal and pathophysiological, occurring through the whole reproductive cycle, while the expression patterns that they exhibit, together with their mutual relations, might serve as factors to better understand the ways in which granulosa cells behave in vitro and possibly in vivo

  • By Affymetrix® Human HgU 219 Array (Affymetrix, Santa Clara, CA, USA), we examined the expression of 22,480 transcripts

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Summary

Introduction

The increase of knowledge about the processes underlining the development of human gametes have brought into light the unique interactions between the tissues involved in that process. Granulosa cells, being a part of almost every stage of folliculo and oogenesis, are deeply involved in storage and maturation of the oocytes, expressing a range of reciprocal interactions with the female gamete [1,2,3]. They play a major role in synthesis, expression, and metabolism of a range of hormones, that function in maintenance of gamete maturation, regulation of ovulation, and sustenance of pregnancy, and perform a wide range of secondary functions defining almost every aspect of physiology associated with reproduction [4]. While the functional analyses of those processes have been long performed and provided us with wide understanding of the ways in which granulosa cells function both by themselves and in relation to the developing oocytes, the knowledge about their exact molecular basis is relatively small [9]

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