Assessment of stress gene mRNAs (HSP-27, 60 and 70) in obstructed rabbit urinary bladder using a semi-quantitative RT-PCR method.
Stress proteins (HSPs) participate in the cellular response to various stresses including hyperthermia, hypoxia and injury. A previous work using northern blot analysis demonstrated increased expression of stress protein 70 (HSP-70) in rabbit bladder tissue subjected to partial outlet obstruction. In order to determine if the increased expression was specific for HSP-70 or, alternatively, indicated a generalized stress protein response, a modified quantitative RT-PCR technique was used to quantitate HSP mRNAs (HSP-27, 60, and 70) in normal and obstructed rabbit urinary bladder tissues. The results show the following: 1) The modified semi-quantitative RT-PCR is a sensitive and reproducible technique for detecting mRNA in bladder tissue. 2) Constitutive levels of HSP-27, HSP-60, and HSP-70 mRNAs were detected in control bladder tissues; the relative signal intensity was highest for HSP-70 and lowest for HSP-27. 3) A transient increase in HSP mRNAs was observed after obstruction; the mRNAs of HSP-27, 60 and 70 increased 4.3-, 5.6-, and 2.4-fold, respectively, at 24 h following obstruction, then gradually returned to control levels by the end of one week post-obstruction and remained stable up to 14 days post-obstruction. These data indicate that the modified quantitative RT-PCR is a useful technique for detecting mRNA in bladder tissue; the stress response which occurs in rabbit urinary bladder tissue following partial outlet obstruction is a general phenomenon.
D000818:Animals; D018834:Chaperonin 60 / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006984:Hypertrophy; D008026:Ligation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D001743:Urinary Bladder / Q000378:metabolism / Q000473:pathology; D001748:Urinary Bladder Neck Obstruction / Q000235:genetics* / Q000378:metabolism
D018834:Chaperonin 60; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D012333:RNA, Messenger