Differential subcellular localization of neural isoforms of the catalytic subunit of calmodulin-dependent protein phosphatase (calcineurin) in central nervous system neurons: immunohistochemistry on formalin-fixed paraffin sections employing antigen retrieval by microwave irradiation.
We examined the immunohistochemical distribution of the two mammalian isoforms of calcineurin catalyic subunits, A alpha and A beta, in central nervous system (CNS) tissues of cows, rats, and humans. Cryostat sections and paraffin sections of parformaldehyde-fixed tissues were stained with antipeptide antibodies for each isoform. The same localization pattern was observed in both cryostat and paraffin sections. In the latter, the intensity of the staining was dramatically enhanced by microwave irradiation. Calcineurin isoforms were localized in a variety of nerve cells but not in neuroglial cells. Their differential expression as the A alpha isoform in the nucleus and the A beta isoform in the cytoplasm was present in a variety of CNS nerve cells, most distinctively in Purkinje cells of the cerebellum and pyramidal cells of the cerebrum, irrespective of species. These results suggest that each isform has distinct intracellular sites of action in CNS neurons and that the phenomenon has been conserved during mammalian evolution.
Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, Japan.
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N
Usuda
H
H
Arai
H
H
Sasaki
T
T
Hanai
T
T
Nagata
T
T
Muramatsu
R L
RL
Kincaid
S
S
Higuchi
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000032:analysis; D019703:Calcineurin; D002148:Calmodulin-Binding Proteins / Q000032:analysis*; D002490:Central Nervous System / Q000737:chemistry*; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008872:Microwaves; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D010749:Phosphoprotein Phosphatases / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D016707:Tissue Fixation
D000941:Antigens; D002148:Calmodulin-Binding Proteins; D005557:Formaldehyde; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases