Expression of carbonic anhydrase II gene in early brain cells as revealed by in situ hybridization and immunohistochemistry.
A mouse carbonic anhydrase (CA II) complementary(c) DNA probe was used for in situ hybridization on mouse brain cultured cells in order to follow CA II gene expression during brain development. An improved method was established using biotinated probes that resulted in a high sensitivity and an absence of background; this method could be combined with immunohistochemistry. Hypothalamic cells of embryonic day (ED) 12-14 mice were cultured for various periods. Chronologic appearance of CA II messenger(m)RNA and protein was studied. The CA II gene transcripts are detectable as early as ED 12-13, although the protein they encode is not detectable until ED 17-18. Gene expression is restricted to 0.1% of the total population. Northern blot analysis confirmed the presence of CA II transcripts in embryonic hypothalamus. At postnatal stage, the majority of glial cells express both the CA II mRNA and the protein. Our results favour the early appearance of a glial lineage in a precise area of the developing CNS. The precocity of CA II gene transcription makes in situ hybridization an invaluable approach in defining the onset of nerve cell lineages during embryonic development.
De Vitry
F
F
Collège de France, Paris.
Gomes
D
D
Rataboul
P
P
Dumas
S
S
Hillion
J
J
Catelon
J
J
Delaunoy
J P
JP
Tixier-Vidal
A
A
Dupouey
P
P
D000818:Animals; D015152:Blotting, Northern; D002256:Carbonic Anhydrases / Q000235:genetics*; D002478:Cells, Cultured; D005796:Genes; D007031:Hypothalamus / Q000196:embryology / Q000201:enzymology; D007527:Isoenzymes / Q000235:genetics*; D051379:Mice; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D014158:Transcription, Genetic
D007527:Isoenzymes; D012333:RNA, Messenger; D002256:Carbonic Anhydrases