Housekeeping gene expression during fetal brain development in the rat-validation by semi-quantitative RT-PCR.
Mammalian gene expression is usually carried out at the level of mRNA where the amount of mRNA of interest is measured under different conditions such as growth and development. It is therefore important to use a "housekeeping gene", that does not change in relative abundance during the experimental conditions, as a standard or internal control. However, recent data suggest that expression of some housekeeping genes may vary with the extent of cell proliferation, differentiation and under various experimental conditions. In this study, the expression of various housekeeping genes (18S rRNA [18S], glyceraldehydes-3-phosphate dehydrogenase [G3PDH], beta-glucuronidase [BGLU], histone H4 [HH4], ribosomal protein L19 [RPL19] and cyclophilin [CY]) was investigated during fetal rat brain development using semi-quantitative RT-PCR at 16, 19 and 21 days gestation. It was found that all genes studied, with exception to G3PDH, did not show any change in their expression levels during development. G3PDH, on the other hand, showed increased expression with development. These results suggest that the choice of a housekeeping gene is critical to the interpretation of experimental results and should be modified according to the nature of the study.
Al-Bader
Maie Dawoud
MD
Department of Physiology, Faculty of Medicine, Kuwait University, P.O. Box 24923, Safat, Zip Code 13110, Kuwait. albader@hsc.edu.kw
Al-Sarraf
Hameed Ali
HA
D000818:Animals; D015152:Blotting, Northern; D001921:Brain / Q000196:embryology / Q000378:metabolism*; D021983:Cyclophilins; D004622:Embryo, Mammalian; D005260:Female; D015870:Gene Expression / Q000502:physiology; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005966:Glucuronidase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D006657:Histones; D008297:Male; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism*; D012337:RNA, Ribosomal, 18S / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012269:Ribosomal Proteins
D006657:Histones; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D012269:Ribosomal Proteins; C050482:Rpl19 protein, rat; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005966:Glucuronidase; D021983:Cyclophilins