title: New insights on flow-independent mechanisms of 99mTc-HMPAO retention in nervous tissue: in vitro study.
abstract: SPECT using 99mTc-hexamethyl propyleneamine oxime (HMPAO) mainly reflects regional cerebral blood flow, however metabolic abnormalities also affect the retention of 99mTc-HMPAO.
  
  
  METHODS
  To rule out any flow factor, a test-tube model was used to evaluate the effects of metabolic alterations both on intracellular trapping of 99mTc-HMPAO and on extracellular glutamate and lactate dehydrogenase (LDH) outflow from rat brain slices.
  
  
  RESULTS
  Under control conditions, slices took up 7.0%+/-1.4% of 99mTc-HMPAO contained in the medium, whereas prelabeled slices released 10.8%+/-2.6% of their radioactive content; glutamate and LDH outflow were 49.1+/-21.6 pmol/mg protein/ min and 4.8+/-0.9 U/L/mg protein/min, respectively. The control medium was altered by adding a metabolic poison (5 mmol/L azide), removing glucose and replacing O2 with N2 to mimic ischemia (in vitro ischemia) and replacing Krebs solution with hypotonic medium to evoke cell lysis. Both azide and in vitro ischemia induced a significant increase in 99mTc-HMPAO release (15.8%+/-3.3% and 18.3%+/-6.2%, respectively), without any modification in LDH efflux. However, only azide reduced the uptake of the tracer. Conversely, glutamate outflow was massive during in vitro ischemia and was far lower during azide treatment. Under hypotonic medium conditions, the release of 99mTc-HMPAO, glutamate and LDH were dramatically increased. Surprisingly, a two-fold increase of 99mTc-HMPAO uptake was also found. When 1 mmol/L glutathione was added to the medium, to convert native lipophilic 99mTc-HMPAO into hydrophilic derivatives, tracer uptake was inhibited both under control and hypotonic medium conditions.
  
  
  CONCLUSIONS
  This study provides evidence that not only poisoning of the tissue but also in vitro ischemia induced a reduction of 99mTc-HMPAO retention. Moreover, we demonstrated that injuries causing cell membrane disruption led to hyperfixation of 99mTc-HMPAO.
authors.affiliation: Department of Experimental and Clinical Medicine, University of Ferrara, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Colamussi
authors.forename: G
authors.initials: G
authors.lastname: Calò
authors.forename: S
authors.initials: S
authors.lastname: Sbrenna
authors.forename: L
authors.initials: L
authors.lastname: Uccelli
authors.forename: C
authors.initials: C
authors.lastname: Bianchi
authors.forename: C
authors.initials: C
authors.lastname: Cittanti
authors.forename: A
authors.initials: A
authors.lastname: Siniscalchi
authors.forename: M
authors.initials: M
authors.lastname: Giganti
authors.forename: R
authors.initials: R
authors.lastname: Roveri
authors.forename: A
authors.initials: A
authors.lastname: Piffanelli
mesh_terms: D000818:Animals; D001386:Azides / Q000494:pharmacology; D001921:Brain / Q000378:metabolism*; D002545:Brain Ischemia / Q000378:metabolism; D002560:Cerebrovascular Circulation; D018698:Glutamic Acid / Q000378:metabolism; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019690:Technetium Tc 99m Exametazime / Q000493:pharmacokinetics*
chemical_list: D001386:Azides; D019275:Radiopharmaceuticals; D019690:Technetium Tc 99m Exametazime; D018698:Glutamic Acid; D007770:L-Lactate Dehydrogenase