Meyer DL et al. (1997), Differential labelling of primary olfactory sys...

XB-ART-16262

Dev Biol 1997 Jul 11;7621-2:275-80.

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Differential labelling of primary olfactory system subcomponents by SBA (lectin) and NADPH-d histochemistry in the frog Pipa.

Meyer DL , Fackler IR , Jadhao AG , D'Aniello B , Kicliter E .

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SBA and NADPH-d histochemistries allow identification of functionally distinct components of the amphibian primary olfactory system. In Pipa, a secondarily aquatic frog, combination of both methodologies, using alternate sets of histological sections, reveals that, apart from Jacobson's organ, this species has a "water-nose" and an "air-nose". The epithelia occupy separate chambers of the olfactory organ and give rise to olfactory nerve fiber bundles that are identified by the dual staining procedure.

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Fig. 1. Peripheral olfactory system of Pipa. A: transverse section through the head showing different components of the olfactory organ after NADPH-d histochemistry. Medial diÕerticulum Ž . Ž . Ž. md; ‘‘air-nose’’ , lateral diÕerticulum ld; ‘‘water-nose’’ , and Jacobson’s organ Jo are visible. Note: large glands Ž . Ž. high magnification is given in A are present exclusively in md; nerve fibers with strong reactivity depicted by arrowheads-asterisks emerge only from 1 ld; only in ld high magnification is given in A staining of the mucosa is massive in all layers. Rhombs label accessory olfactory tract; d, dorsal; l, Ž . 2 lateral. B: adjacent section to A processed with soybean agglutinin histochemistry. Fibers from ld asterisk and Jo rhombs are stained. Similarity of Ž. Ž . Ž . mucosal staining in ld B and in Jo B is shown at high magnification. Calibration bars in A; also applies to B Ž. Ž. Ž . Ž s500 mm; in A ; also applies to A , 1 2 1 2 B , and B .s50 mm.

Fig. 2. A: paramedial sagittal section of the head of Pipa stained with HRP-labelled SBA. To the right, the medial diÕerticulum Ž‘‘air-nose’’; md, bold ‘‘x’’ is visible. The peripheral portion of fibers emerging from md is located in more lateral planes of sectioning. Half way to the olfactory bulb, the . practically unstained md projections depicted by ‘‘ Ž . x’’s are positioned more medial and, thus, are visible in the photomicrograph. They terminate in the dorsal olfactory bulb. By contrast, fibers from the lateral diÕerticulum Ži.e. the ‘‘water nose’’; located further rostral than the area covered by this photomicrograph are reactive in SBA histochemistry. Where present in this section, they are marked by asterisks. They innervate the rostro-ventral portion . of the olfactory bulb left large arrow . The darkest staining is displayed by fibers of the accessory olfactory system Jacobson’s organ is not shown . Ž . Ž . Where visible, these fibers are depicted by rhombs. Tel, telencephalon. B: SBA reacted transverse section of the left olfactory nerve d, dorsal; m, medial . Ž . Two stained and one unstained component can be discerned. The unstained bundle Ž. Ž . x originates in md. The larger one asterisk of the reactive bundles contains fibers from lateral diÕerticulum, the smaller and more lateral one rhomb is the accessory olfactory nerve. Its staining is particularly dark. C: Ž . NADPH-d reacted transverse section of the left olfactory nerve; adjacent section to B ; d, dorsal; m, medial . Two unstained and one stained component Ž.Ž . can be discerned. The massively stained bundle asterisk emerges from the Ž . Ž. lateral diÕerticulum. All fibers from Jacobson’s organ rhomb are devoid of reactivity. Most fibers of the third bundle from md; Ž . x are unstained. However, a small percentage of the fibers located at the lateral edge of this bundle Ž . arrow is reactive in NADPH-d histochemistry. Possibly, this subcomponent is also stained in the SBA material but it cannot be discerned with certainty due to reactivity of the meninges; compare B . The region of interest is given at higher magnification in C ; two arrows. D: SBA reacted transverse Ž. Ž .1 section of the left olfactory and accessory olfactory bulbs. The accessory olfactory bulb rhomb-arrow as well as the terminal fields of projections from Ž . lateral diÕerticulum Ž . asterisk-arrow are labelled. E: NADPH-d reacted transverse section of the right olfactory and accessory olfactory bulbs; adjacent section to D . The accessory olfactory bulb rhomb-arrow is weakly stained whereas the terminal fields of projections from Ž. Ž . lateral diÕerticulum Ž . Ž. Ž . Ž . Ž . asterisk-arrow are strongly labelled. Calibration bars in A s1 mm; in C; also applies to B s75 mm; in C1 s50 mm; in D; also applies to E s300 mm.