Cell-specific transcriptional profiling reveals candidate mechanisms regulating development and function of uterine epithelia in mice

Abstract

All mammalian uteri have luminal (LE) and glandular epithelia (GE) in their endometrium. The LE mediates uterine receptivity and blastocyst attachment for implantation, and the GE synthesize and secrete or transport bioactive substances involved in blastocyst implantation, uterine receptivity, and stromal cell decidualization. However, the mechanisms governing uterine epithelial development after birth and their function in the adult are not fully understood. Here, comprehensive microarray analysis was conducted on LE and GE isolated by laser capture microdissection from uteri on Postnatal Day 10 (PD 10) and day of pseudopregnancy (DOPP) 2.5 and 3.5. This data was integrated with analysis of uteri from gland-containing control and aglandular progesterone-induced uterine gland knockout mice from PD 10 and DOPP 3.5. Many genes were expressed in both epithelia, but there was greater expression of genes in the LE than in the GE. In the neonate, GE-expressed genes were enriched for morphogenesis, development, migration, and retinoic acid signaling. In the adult, LE-expressed genes were enriched for metabolic processes and steroid biosynthesis, whereas retinoid signaling, tight junction, extracellular matrix, and regulation of kinase activity were enriched in the GE. The transcriptome differences in the epithelia support the idea that each cell type has a distinct and complementary function in the uterus. The candidate genes and regulatory networks identified here provide a framework to discover new mechanisms regulating development of epithelia in the postnatal uterus and their functions in early pregnancy.

Keywords: endometrium; gene expression; guinea pigs; implantation; mice; rodents (rats; uterus; voles).

Fig. 1

Quantitative PCR validation of selected genes identified by LCM and microarray analysis. The mRNA levels of the indicated genes were measured in microdissected luminal epithelial (LE) and glandular epithelial (GE) cells of the uteri from PD 10, DOPP 2.5, and DOPP 3.5 mice by semiquantitative RT-PCR analysis (n = 4 mice/cell type/day). Data are presented as fold change of target mRNA levels in GE as compared to LE.

Fig. 2

Luminal epithelial (LE) genes expressed in the neonatal and adult mouse uteri.A) Heat map of the most LE-enriched genes in PD 10, DOPP 2.5, and DOPP 3.5. Normalized probe intensity values are presented for the top 30 genes significantly enriched in LE.B) Venn diagram comparing LE-enriched genes between neonatal (PD 10) and adult uteri (DOPP 2.5 and 3.5).

Fig. 3

Glandular epithelial (GE) genes expressed in neonatal and adult mouse uteri.A) Heat map of the most GE-enriched genes in PD 10, DOPP 2.5, and DOPP 3.5. Normalized probe intensity values are presented for the top 30 genes significantly enriched in GE.B) Venn diagram comparing GE-enriched genes between neonatal (PD 10) and adult uteri (DOPP 2.5 and 3.5).

Fig. 4

Genes enriched in the uterine glands of neonatal mice.A) Venn diagram is presented showing intersection between GE-enriched genes and genes increased in the uteri of control as compared to PUGKO mice on PD 10 as determined by microarray analysis. Genes marked in red were validated by RT-PCR analysis.B) Validation of selected GE-enriched genes by semiquantitative RT-PCR analysis. Relative mRNA levels of the indicated genes were measured in the uteri of control and PUGKO mice on PD 10 (n = 4 mice/day/treatment). Real-time PCR data are presented as fold change relative to the mRNA level in uteri from control mice.

Fig. 5

Genes enriched in the uterine glands of adult pseudopregnant mice.A) Venn diagram is presented showing intersection between GE-enriched genes and genes increased in the uteri of control as compared to PUGKO mice on DOPP 3.5 as determined by microarray gene expression analysis. Genes marked in red were validated by RT-PCR analysis.B) Validation of selected GE-enriched genes by semiquantitative RT-PCR analysis. Relative mRNA levels of the indicated genes were measured in the uteri of control and PUGKO mice on DOPP 3.5 (n = 4 mice/day/treatment). Real-time PCR data are presented as fold change relative to the mRNA level in uteri from control mice.