Evidence for two isoforms of carbonic anhydrase II in the leech (Hirudo medicinalis) central nervous system

1. We dissected, homogenized and prepared ganglia and connectives from the central nervous system of medicinal leeches for SDS gel electrophoresis. The isolated proteins were transferred to nitrocellulose and incubated with affinity column-purified antibodies. 2. The immunoblots showed a strong positive reaction of a bovine carbonic anhydrase standard at a molecular weight of 29 kDa, and a distinct double-bond at the same molecular weight in the analyzed material. 3. We demonstrated with rat antibodies that carbonic anhydrase II is detectable in the leech central nervous system as the main isoenzyme. 4. The biochemical knowledge of carbonic anhydrase reported here agrees well with the immunocytochemical locations, thus affirming the validity of specific staining.