negative inference; confirmed by additional inferences
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EXPRESSION: negative (inference)
ANIMAL: rat
PROTOCOL: immunohistochemistry
Page location: p1820, left mid
Linking PMID: 730852 Linking Quote: Citation included in evidentiary quote.
"Here we examined whether interneurons were among the hilar neurons that express CGRP [calcitonin gene-related peptide], or this neuropeptide is present exclusively in mossy cells. The mirror technique of Kosaka et al. (1985) was applied to hippocampal sections from three colchicine-treated animals. A total of 60, 62, 77 and 75 CGRP-positive neurons with somata cut in half on the surface of the sections were identified, and their corresponding other soma-halves located in the adjacent sections immunostained for PV [parvalbumin], CR [calretinin], CCK [cholecystokinin], or SOM [somatostatin] respectively. None of the examined CGRP-containing neurons was immunoreactive for any calcium binding proteins or neuropeptides (Fig. 8). In a recent study (Leranth et al., 1996) GluR2/3 [glutamate receptor 2/3] immunoreactivity was shown to be absent from interneurons in the dentate gyrus, but present in numerous hilar cells, which were suggested to correspond to mossy cells. In order to test this hypothesis, and also to have a positive finding in our colocalization study, we examined the coexistence of CGRP and GluR2/3 using the same protocol as above (Fig. 9). Out of 105 CGRP-immunoreactive neurons (from four animals) 101 were unequivocally positive for GluR2/3, and only four were found to be negative. ... [p1820, right middle] The light microscopic features of CGRP-positive cells, e.g. the shape and size of somata, the prominent proximal dendrites covered by thorny excrescences, and the limitation of the dendritic tree to the hilar region, all closely resemble the characteristics of mossy cells described in earlier Golgi or intracellular labelling studies (Amaral, 1978; Ribak et al., 1985; Scharfman et al., 1990; Frotscher et al., 1991; Buckmaster et al., 1992, 1996; Soltesz et al., 1993)."
Interpretation notes: Glutamatergic and GABAergic CR+ types in DG Smo_Smi_SG_H are probably CGRP- by contrapositive logic; Clopper-Pearson confidence interval lower bound = 94.2% (n = 62/62)
EXPRESSION: negative (inference)
ANIMAL: rat
PROTOCOL: immunohistochemistry
Page location: p1820, left mid
Linking PMID: 730852 Linking Quote: Citation included in evidentiary quote.
"Here we examined whether interneurons were among the hilar neurons that express CGRP [calcitonin gene-related peptide], or this neuropeptide is present exclusively in mossy cells. The mirror technique of Kosaka et al. (1985) was applied to hippocampal sections from three colchicine-treated animals. A total of 60, 62, 77 and 75 CGRP-positive neurons with somata cut in half on the surface of the sections were identified, and their corresponding other soma-halves located in the adjacent sections immunostained for PV [parvalbumin], CR [calretinin], CCK [cholecystokinin], or SOM [somatostatin] respectively. None of the examined CGRP-containing neurons was immunoreactive for any calcium binding proteins or neuropeptides (Fig. 8). In a recent study (Leranth et al., 1996) GluR2/3 [glutamate receptor 2/3] immunoreactivity was shown to be absent from interneurons in the dentate gyrus, but present in numerous hilar cells, which were suggested to correspond to mossy cells. In order to test this hypothesis, and also to have a positive finding in our colocalization study, we examined the coexistence of CGRP and GluR2/3 using the same protocol as above (Fig. 9). Out of 105 CGRP-immunoreactive neurons (from four animals) 101 were unequivocally positive for GluR2/3, and only four were found to be negative. ... [p1820, right middle] The light microscopic features of CGRP-positive cells, e.g. the shape and size of somata, the prominent proximal dendrites covered by thorny excrescences, and the limitation of the dendritic tree to the hilar region, all closely resemble the characteristics of mossy cells described in earlier Golgi or intracellular labelling studies (Amaral, 1978; Ribak et al., 1985; Scharfman et al., 1990; Frotscher et al., 1991; Buckmaster et al., 1992, 1996; Soltesz et al., 1993)."
Interpretation notes: Glutamatergic and GABAergic SOM+ types in DG Smo_Smi_SG_H are probably CGRP- by contrapositive logic; Clopper-Pearson confidence interval lower bound = 95.2% (n = 75/75)
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negative inference; confirmed by additional inferences