Assessment of stress gene mRNAs (HSP-27, 60 and 70) in obstructed rabbit urinary bladder using a semi-quantitative RT-PCR method

Mol Cell Biochem. 1995 Jul 5;148(1):1-7. doi: 10.1007/BF00929496.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Chaperonin 60 / biosynthesis*
  • Chaperonin 60 / genetics
  • Gene Expression Regulation
  • HSP70 Heat-Shock Proteins / biosynthesis*
  • HSP70 Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / biosynthesis*
  • Heat-Shock Proteins / genetics
  • Hypertrophy
  • Ligation
  • Polymerase Chain Reaction*
  • RNA, Messenger / biosynthesis*
  • RNA, Messenger / genetics
  • Rabbits
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Urinary Bladder / metabolism
  • Urinary Bladder / pathology
  • Urinary Bladder Neck Obstruction / genetics*
  • Urinary Bladder Neck Obstruction / metabolism

Substances

  • Chaperonin 60
  • HSP70 Heat-Shock Proteins
  • Heat-Shock Proteins
  • RNA, Messenger