PMID- 2312356
OWN - NLM
STAT- MEDLINE
DCOM- 19900423
LR  - 20190828
IS  - 0301-5564 (Print)
IS  - 0301-5564 (Linking)
VI  - 93
IP  - 3
DP  - 1990
TI  - Microphotometric determination of enzymes in brain sections. I. Hexokinase.
PG  - 295-8
AB  - A histochemical procedure was established for the microphotometric determination 
      of hexokinase (HK) in sections of the rat hippocampus, which served as an 
      exemplary brain region. For this quantitative procedure, slides were coated with 
      glucose 6-phosphate dehydrogenase (G6PDH) as an auxiliary enzyme and sections 
      were mounted onto this enzyme film. The sections were then incubated with the 
      following adapted incubation medium: 5 mM D-glucose, 1.5 mM NADP, 7.5 mM ATP, 4 
      mM nitroblue tetrazolium chloride, 10 mM NaN3, 10 mM MgCl2, 0.25 mM phenazine 
      methosulfate, 1 U/ml G6PDH, 22% polyvinyl alcohol in 0.05 M Hepes buffer; the 
      final pH was 7.5. A linear response of the reaction was observed in the initial 
      10 min of reaction (kinetic and end-point measurements). The relationship between 
      HK activity and section thickness was linear up to 5 microns. The need for such 
      thin sections is discussed in relation to the limited penetration of the 
      auxiliary enzyme into the section. It is concluded that the quantitative 
      demonstration of HK in brain sections could be a valuable tool for studying the 
      local metabolic entrance of glucose in the glycolytic pathway.
FAU - Kugler, P
AU  - Kugler P
AD  - Department of Anatomy, University of Wurzburg, Federal Republic of Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Histochemistry
JT  - Histochemistry
JID - 0411300
RN  - 0 (Indicators and Reagents)
RN  - EC 1.1.1.49 (Glucosephosphate Dehydrogenase)
RN  - EC 2.7.1.1 (Hexokinase)
SB  - IM
MH  - Animals
MH  - Glucosephosphate Dehydrogenase
MH  - Hexokinase/*analysis
MH  - Hippocampus/*enzymology
MH  - Histocytochemistry/methods
MH  - Indicators and Reagents
MH  - Male
MH  - Rats
MH  - Rats, Inbred Strains
EDAT- 1990/01/01 00:00
MHDA- 1990/01/01 00:01
CRDT- 1990/01/01 00:00
PHST- 1990/01/01 00:00 [pubmed]
PHST- 1990/01/01 00:01 [medline]
PHST- 1990/01/01 00:00 [entrez]
AID - 10.1007/BF00266391 [doi]
PST - ppublish
SO  - Histochemistry. 1990;93(3):295-8. doi: 10.1007/BF00266391.