Alkaline phosphatase activity in the brain of the American cockroach Periplaneta americana L.
The supra- and suboesophageal ganglia of the American cockroach contain material which catalyses the alkaline hydrolysis (pH 9.5) of 5-bromo-4-chloro-3-indolyl phosphate in the presence of Nitro blue tetrazolium. Histochemical studies on unfixed cryostat sections indicate that this type of alkaline phosphatase is restricted to discrete regions in the cockroach brain. Highest enzyme activity is encountered in the mushroom bodies, central body, antennal glomeruli and specific parts of some distinct neural connections including the optic nerve, antennal nerve, circumoesophageal connectives and nerves leaving the suboesophageal ganglion. Tissue fixation by use of formaldehyde-type fixatives, as well as routine paraffin-embedding, completely destroy all histochemically detectable enzyme activity. Native polyacrylamide gradient electrophoresis suggests that the alkaline phosphatase activity is present as multiple isozymic forms, which show up in the 120-130 kD range of standard proteins. Enzyme activity becomes undetectable after fixation (trichloroacetic acid, formaldehyde containing fixatives) of electrophoretically separated native proteins, as well as after electrophoresis in denaturing conditions (SDS and beta-mercapto-ethanol, boiling). However, the enzyme activity remains virtually unaffected after storage of the sample for prolonged periods at -20 to -80 degrees C.
Verhaert
P D
PD
Department of Biology, University of Waterloo, Ontario, Canada.
Walgraeve
H R
HR
Downer
R G
RG
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000033:anatomy & histology / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D008297:Male; D009420:Nervous System / Q000201:enzymology; D009490:Neurosecretory Systems / Q000201:enzymology; D010232:Paraffin; D010526:Periplaneta / Q000201:enzymology*; D013997:Time Factors; D014018:Tissue Distribution
D010232:Paraffin; D000469:Alkaline Phosphatase