Methods for evaluating histocompatibility antigen gene expression in the baboon.
A wide variety of techniques has been developed for qualitative and quantitative analysis of gene expression in human cells and tissues. Two commonly used methods are reverse-transcription (RT)-polymerase chain reaction (PCR) to analyze the transcribed messenger RNAs (mRNA) and immunohistochemistry to detect the translated proteins. These techniques can be modified and adapted for use in analyzing gene expression in animal models. In particular, as a result of the close phylogenetic relationship between humans and nonhuman primates, human reagents, especially antibodies, cross-react with nonhuman primate tissues. However, the results are not always satisfactory as some antibodies may cross-react with irrelevant antigens in these tissues. In this chapter, we describe the use of RT-PCR and immunohistochemical techniques to analyze expression of Paan-AG, a novel class lb major histocompatibility complex antigen in the olive baboon (Papio anubis) placenta. We used Paan-AG-specific primers to amplify Paan-AG transcripts from baboon placenta, and generated Paan-AG isoform-specific polyclonal antibodies for use in immunohistochemistry.
Langat
Daudi K
DK
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, USA.
Fazleabas
Asgerally T
AT
Hunt
Joan S
JS
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000917:Antibody Formation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015870:Gene Expression; D015395:Histocompatibility Antigens Class I / Q000032:analysis*; D007150:Immunohistochemistry / Q000379:methods*; D023421:Models, Animal; D048530:Papio anubis / Q000276:immunology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods
D000906:Antibodies; D015395:Histocompatibility Antigens Class I; D012333:RNA, Messenger