RNAlater® Tissue Collection
RNAlater® Tissue Collection: RNA Stabilization Solution is an aqueous tissue storage reagent that stabilizes and protects cellular RNA in intact, unfrozen tissue samples. RNAlater® Solution eliminates the need to immediately process tissue samples or to freeze samples in liquid nitrogen for later processing. Thus, you are left with more time for sample collection without jeopardizing RNA expression profiles in your samples. Here we describe how three different research groups used RNAlater® Solution to preserve samples for microarray and RT-PCR studies.
Perfusion-fixation of Central and Peripheral Nervous System Samples
Storage of Placental and Embryonic Samples
Aspartyl-(asparaginyl) β-hydroxylase (AAH) is a transmembrane protein that hydroxylates epidermal growth factor-like domains of proteins that have a functional role in cell motility and invasion. Dr. Fusun Gundogan and colleagues (Brown Medical School, Providence, RI) characterized the potential role of AAH in implantation using human placentas from terminated pregnancies, normal term deliveries, spontaneous abortuses, and small-for-gestational-age term deliveries . Snap-frozen or RNAlater solution-preserved specimens were used for RNA analysis of AAH expression by RT-PCR and protein analysis by Western blotting. Immunohistochemical staining and RNA analysis suggested that AAH may serve as a biomarker of impaired implantation.
The process of testicular descent depends on the gubernaculum, which enlarges and migrates into the scrotum, creating a space into which the testis descends. Development of the fetal gubernaculum requires the expression of leucine rich G protein coupled receptor and its ligand, insulin-like 3. Dr. Julia Barthold and colleagues (A I DuPont Hospital for Children, Wilmington, DE) compared expression levels of these two genes in Long-Evans orl and wild type rats during perinatal development—failure of testicular descent occurs in ~65% of spontaneously cryptorchid Long-Evans orl rats . Males from both strains obtained at gestational days 18–21 and day of birth were preserved in RNAlater Solution for at least 24 hours. The size and position of the testes, kidneys, and gubernacula were determined by microdissection and image analysis. Leucine-rich G protein coupled receptor and insulin-like 3 mRNA expression were analyzed by RT-PCR. Data suggest that the impaired shortening of the Long-Evans orl gubernaculum may interfere with the timing and quality of its inversion at birth, leading to failure of testicular descent in some newborn males.
- LeDoux MS, Xu L, Xiao J, Ferrell B, Menkes DL, Homayouni R (2006) Murine central and peripheral nervous system transcriptomes: comparative gene expression. Brain Res 1107:24–41.
- Gundogan F, Elwood G, Greco D, Rubin LP, Pinar H, Carlson RI, Wands JR, de la Monte SM (2007) Role of aspartyl-(asparaginyl) beta-hydroxylase in placental implantation: relevance to early pregnancy loss. Hum Pathol 38:50–59.
- Barthold JS, Si X, Stabley D, Sol-Church K, Campion L, McCahan SM (2006) Failure of shortening and inversion of the perinatal gubernaculum in the cryptorchid long-evans orl rat. J Urol 176:1612–1617