Analysis of segmental renal gene expression by laser capture microdissection

Y. Kohda, H. Murakami, O. W. Moe, R. A. Star

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

Background. The study of normal renal physiology has been greatly aided by microdissection techniques that have delineated the exceptional functional and cellular heterogeneity both along the nephron and between different nephron populations. These techniques are not widely used to study renal injury as microdissection is difficult because of tissue necrosis or fibrosis. We developed a procedure to detect specific gene expression in specific locations of the kidney in histologic sections. Methods. The anatomic specificity of laser capture microdissection (LCM) was employed with the sensitivity of reverse transcriptase-polymerase chain reaction (RT-PCR). Results. LCM/RT-PCR detected mRNA for podoplanin in 2% of a single glomerulus, rat basic amino acid transporter in 6% of a single cross-section of proximal straight tubule, and renin in eight proximal convoluted tubule cross-sections. LCM/RT-PCR could isolate pure populations of proximal convoluted tubules, proximal straight tubules, and thick ascending limbs from renal histologic sections, although pure collecting ducts could not be isolated. LCM/RT-PCR localized ischemia-reperfusion-induced induction of KC/interleukin-8 primarily to the medullary thick ascending limb, and detected transforming growth factor-β (TGF-β) mRNA in glomeruli of a patient with membranous glomerulonephropathy. Conclusions. When used with an appropriate laser spot size, LCM/RT-PCR can measure gene expression in glomeruli or specific parts of the nephron and can study alterations in steady-state mRNA levels in animal models of renal disease. The applications, limitations, and refinements of this approach are discussed.

Original languageEnglish (US)
Pages (from-to)321-331
Number of pages11
JournalKidney International
Volume57
Issue number1
DOIs
StatePublished - 2000

Fingerprint

Laser Capture Microdissection
Reverse Transcriptase Polymerase Chain Reaction
Nephrons
Kidney
Gene Expression
Microdissection
Messenger RNA
Extremities
Amino Acid Transport Systems
Animal Disease Models
Basic Amino Acids
Membranous Glomerulonephritis
Transforming Growth Factors
Interleukin-8
Renin
Population
Reperfusion
Lasers
Fibrosis
Necrosis

Keywords

  • Fibrosis
  • Necrosis
  • Nephron
  • Renal injury
  • Tissue microdissection

ASJC Scopus subject areas

  • Nephrology

Cite this

Analysis of segmental renal gene expression by laser capture microdissection. / Kohda, Y.; Murakami, H.; Moe, O. W.; Star, R. A.

In: Kidney International, Vol. 57, No. 1, 2000, p. 321-331.

Research output: Contribution to journalArticle

Kohda, Y. ; Murakami, H. ; Moe, O. W. ; Star, R. A. / Analysis of segmental renal gene expression by laser capture microdissection. In: Kidney International. 2000 ; Vol. 57, No. 1. pp. 321-331.
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