Utility of Confocal Laser Scanning Microscopy (CLSM): With Reference to Interpretation in Immunostaining

Ogane, Naoki; Yasuda, Masanori; Hayashi, Hiroyuki; Kameda, Yoichi; Minematsu, Takeo; Itoh, Johbu; Osamura, R. Yoshiyuki

doi:10.1267/ahc.38.267

Cited by 1 publication

(1 citation statement)

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“…PBST was used to dilute the antibodies and wash the sections between each step. The free-floating sections were mounted on gelatin-coated glass slides and examined under a confocal laser-scanning microscope (Bio-Rad, Hercules, CA, USA) as described previously [7, 16]. …”

Section: Methodsmentioning

confidence: 99%

Comparison of FGF1 (aFGF) Expression between the Dorsal Motor Nucleus of Vagus and the Hypoglossal Nucleus of Rat

Toyoda

Okano

Bamba

et al. 2006

Acta Histochem. Cytochem

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Neurons in the dorsal motor nucleus of the vagus (DMNV) are more severely affected by axonal injury than most other nerves, such as those of the hypoglossal nucleus. However, the mechanism underlying such a response remains unclear. In this study, we compared the expression of fibroblast growth factor 1 (FGF1), a neurotrophic factor, between the DMNV and the hypoglossal nucleus by RT-PCR and immunohistochemical analyses. RT-PCR showed that the level of FGF1 mRNA expression in the DMNV was lower than that in the hypoglossal nucleus (P0.01). Immunohistochemistry revealed that FGF1 was localized to neurons. FGF1-positive neurons in large numbers were evenly distributed in the hypoglossal nucleus, whereas FGF1-positive neurons were located in the lateral part of the DMNV. Double immunostaining for FGF1 and choline acetyltransferase demonstrated that 22.7% and 78% of cholinergic neurons were positive for FGF1 in the DMNV and hypoglossal nucleus, respectively. A tracing study with cholera toxin B subunit (CTb) demonstrated that cholinergic neurons sending their axons from the DMNV to the superior laryngeal nerve were FGF1-negative. The results suggest that the low expression of FGF1 in the DMNV is due to severe damage of neurons in the DMNV.

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Section: Methodsmentioning

confidence: 99%