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Journal of Histochemistry and Cytochemistry, Vol. 45, 1147-1156, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

Visualization of ß-Galactosidase by Enzyme and Immunohistochemistry in the Olfactory Bulb of Transgenic Mice Carrying the LacZ Transgene

Gabriela Sekerková1,a,a,c, Zoya Katarovaa, Ferenc Joó2,b, Joachim R. Wolffd, Simona Prodan3,a, and Gábor Szabó
a Institutes of Biochemistry, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
b Biophysics, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
c Neurobiological Institute, Slovakian Academy of Sciences, Kosice, Slovak Republic
d Department of Anatomy, University of Göttingen, Göttingen, Germany

Correspondence to: Gábor Szabó, Lab. of Molecular Neurogenetics, Inst. of Biochemistry, Biological Research Center, Hungarian Academy of Sciences, PO Box 521, Temesvári krt. 62, Szeged, Hungary.

In the olfactory bulb (OB) of a transgenic mouse line that carries the bacterial LacZ gene under the control of the 5'-regulatory region of the GAD67 gene, expression of the ß-galactosidase was confined almost exclusively to the non-GABAergic mitral and tufted cells. By light microscopy, enzyme histochemistry showed strong staining in the cell bodies and faint diffuse staining in the axons and dendrites. With immunohistochemistry for ß-galactosidase the entire cytoplasm, including the axons and dendrites, was strongly stained. By electron microscopy, ß-galactosidase enzyme histochemistry resulted in a submicroscopic reaction product that was diffusely distributed in the cytoplasm of neurons. In addition, large deposits of the reaction product were also seen attached to the cytoplasmic side of the membranes. In contrast, when the intracellular localization of ß-galactosidase was determined by immunohistochemistry, homogeneous cytoplasmic staining was obtained that filled the entire cytoplasm including the terminal dendrites and fine axons. Therefore, synaptic contacts of the ß-galactosidase-positive output neurons with other ß-galactosidase-negative neuronal cells were readily recognized in the OB. As we demonstrated, transgenic mouse lines expressing the LacZ reporter gene in a well-defined neuronal subpopulation can be used to follow ß-galactosidase-positive neurons and to directly identify their synaptic connections. (J Histochem Cytochem 45:1147-1155, 1997)

Key Words: olfactory system, output neurons, neuronal labeling, GAD-LacZ fusion protein, transgenic expression


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A. N. van den Pol and P. K. Ghosh
Selective Neuronal Expression of Green Fluorescent Protein with Cytomegalovirus Promoter Reveals Entire Neuronal Arbor in Transgenic Mice
J. Neurosci., December 15, 1998; 18(24): 10640 - 10651.
[Abstract] [Full Text] [PDF]



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