Transcriptomic Profile of New Gene Markers Encoding Proteins Responsible for Structure of Porcine Ovarian Granulosa Cells.

Jakub Kulus,Maciej Zdun,Wiesława Kranc,Paul Mozdziak,Jędrzej M Jaśkowski,Hanna Piotrowska-Kempisty,Bartosz Kempisty,Magdalena Kulus,Karol Jopek,Małgorzata Józkowiak,Dorota Bukowska,Paweł Antosik

doi:10.3390/biology10111214

Abstract

The extracellular matrix (ECM) in granulosa cells is functionally very important, and it is involved in many processes related to ovarian follicle growth and ovulation. The aim of this study was to describe the expression profile of genes within granulosa cells that are associated with extracellular matrix formation, intercellular signaling, and cell-cell fusion. The material for this study was ovaries of sexually mature pigs obtained from a commercial slaughterhouse. Laboratory-derived granulosa cells (GCs) from ovarian follicles were cultured in a primary in vitro culture model. The extracted genetic material (0, 48, 96, and 144 h) were subjected to microarray expression analysis. Among 81 genes, 66 showed increased expression and only 15 showed decreased expression were assigned to 7 gene ontology groups "extracellular matrix binding", "extracellular matrix structural constituent", "binding, bridging", "cadherin binding", "cell adhesion molecule binding", "collagen binding" and "cadherin binding involved in cell-cell adhesion". The 10 genes with the highest expression (POSTN, ITGA2, FN1, LAMB1, ITGB3, CHI3L1, PCOLCE2, CAV1, DCN, COL14A1) and 10 of the most down-regulated (SPP1, IRS1, CNTLN, TMPO, PAICS, ANK2, ADAM23, ABI3BP, DNAJB1, IGF1) were selected for further analysis. The results were validated by RT-qPCR. The current results may serve as preliminary data for further analyses using in vitro granulosa cell cultures in assisted reproduction technologies, studies of pathological processes in the ovary as well as in the use of the stemness potential of GCs.

Highlights

The extracellular matrix (ECM) is an extremely important structure present in all tissues of the animal organism
For the microarray gene expression analysis of porcine granulosa cells allows the study of the gene expression of 27,558 transcripts at 0, 48, 96, and 144 h of in vitro granulosa cell culture
This paper focused on the genes involved in cadherin and collagen binding and structuralization of extracellular matrix components

Summary

Introduction

The extracellular matrix (ECM) is an extremely important structure present in all tissues of the animal organism. The fact that an extracellular matrix is composed of a variety of components results in a vast array of functions [4] It is a unique microenvironment for cells (including the delivery of osmotic forces), a mechanical support or pathway for the passage of nutrients, hormones and extracellular signals to the target cells as well as regulation of gene expression and cytokine release [1,5]. Heparan sulfate, and others are connected [6,18,19,20] These components may be present in different conforms, composed of various chains, affecting the uniqueness of a specific basal lamina, which in turn affects its functionality [6,21]

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