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Assessment and Application of Laser Microdissection for Analysis of Gene Expression in the Rhesus Monkey Endometrium¹

^a Division of Endocrinology, ^b Department of Obstetrics and Gynecology, ^c Department of Physiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655

We investigated the use of laser capture microdissection (LCM) to identify differences in gene expression between cell types or regions within the rhesus monkey endometrium. Different cell types were harvested from the two major regions of the endometrium during midsecretory phases (Days 21–23) of adequate artificial menstrual cycles: glandular epithelia (G) or stroma (S) from the functionalis (F) or the basalis (B). Amplification of the cDNA populations (primer-specific adaptors) was used to increase the amount of nucleic acid for further analysis. This single amplification step allowed us to detect the housekeeping genes (glyceraldehyde-3-phosphate dehydrogenase and 18S rRNA) and the cDNA smears in the samples. Using differential display reverse transcription polymerase chain reaction (DDRT-PCR), six fragments were selected, cloned, and sequenced based on their regional and cell type localization. Primer-specific PCR analysis subsequently confirmed the localization of three fragments: F1, highly expressed in the functionalis but not the basalis, was homologous (93% identical) to the human leukotriene B4 receptor BLT2; FS-1, highly expressed only in the stroma of the functionalis, had a 94% homology with an as yet uncharacterized gene (FLJ124360); and BG-1, primarily expressed only in the glandular epithelia of the basalis, showed a 98% homology with an uncharacterized bacterial artificial chromosome clone sequence. These LCM-generated cDNA populations coupled with DDRT-PCR can provide an important avenue for the identification of new or novel gene fragments that display cell type- or region-specific gene expression in the rhesus monkey endometrium.

¹ This work was supported in part by grants from the NICHD (HD 31620) and the Worcester Foundation for Biomedical Research. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the NICHD.

² Correspondence: William C. Okulicz, Department of OB/GYN and Physiology, UMass Medical School, 55 Lake Ave. N, Worcester, MA 01655. FAX: 508 856 5933; william.okulicz{at}umassmed.edu

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