Lázár GY et al. (1991), Distribution of galanin-like immunoreactivity i...

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J Comp Neurol 1991 Aug 01;3101:45-67. doi: 10.1002/cne.903100106.

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Distribution of galanin-like immunoreactivity in the brain of Rana esculenta and Xenopus laevis.

Lázár GY , Liposits ZS , Tóth P , Trasti SL , Maderdrut JL , Merchenthaler I .

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The immunocytochemical distribution of galanin-containing perikarya and nerve terminals in the brain of Rana esculenta and Xenopus laevis was determined with antisera directed toward either porcine or rat galanin. The pattern of galanin-like immunoreactivity appeared to be identical with antisera directed toward either target antigen. The distribution of galanin-like immunoreactivity was similar in Rana esculenta and Xenopus laevis except for the absence of a distinct laminar distribution of immunoreactivity in the optic tectum of Xenopus laevis. Galanin-containing perikarya were located in all major subdivisions of the brain except the metencephalon. In the telencephalon, immunoreactive perikarya were detected in the pars medialis of the amygdala and the preoptic area. In the diencephalon, immunoreactive perikarya were detected in the caudal half of the suprachiasmatic nucleus, the nucleus of the periventricular organ, the ventral hypothalamus, and the median eminence. In the mesencephalon, immunoreactive perikarya were detected near the midline of the rostroventral tegmentum, in the torus semicircularis and, occasionally, in lamina A and layer 6 of the optic tectum. In the myelencephalon, labelled perikarya were detected only in the caudal half of the nucleus of the solitary tract. Immunoreactive nerve fibers of varying density were observed in all subdivisions of the brain with the densest accumulations of fibers occurring in the pars lateralis of the amygdala and the preoptic area. Dense accumulations of nerve fibers were also found in the lateral septum, the medial forebrain bundle, the periventricular region of the diencephalon, the ventral hypothalamus, the median eminence, the mesencephalic central gray, the laminar nucleus of the torus semicircularis, several laminae of the optic tectum, the interpeduncular nucleus, the isthmic nucleus, the central gray of the rhombencephalon, and the dorsolateral caudal medulla. The extensive system of galanin-containing perikarya and nerve fibers in the brain of representatives of two families of anurans showed many similarities to the distribution of galanin-containing perikarya and nerve fibers previously described for the mammalian brain.

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Species referenced: Xenopus laevis
Genes referenced: gal.1 gal.2 mhc2-dab

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	Fig. 1. Distribution of galanin-like immunoreactivity in the brain of Rana esculenta. The drawing below E shows the transverse planes chosen for the illustrations. A, B, and C are from the telencephalon, whileD and E are from the diencephalon. The pre tectum is represented by F, while the mesencephalon is represented by G and H. I, J, K, and L are from the rhombencephalon. The location of immunoreactive nerve fibers, terminals, and perikarya is represented in the left half of each drawing, while the neuroanatomical parcellations are designated in the right half. The darkest pattern indicates the sites where the most intense immunoreactivity was detected. Dense patterns of small dots indicate areas where a considerable number of labelled axon terminals were detected. Crosses represent immunoreactive fibers and scattered terminals. Filled triangles represent intensely stained perikarya, while empty triangles indicate faintly stained cells. The three layers of immunoreactive nerve terminals in the optic tectum illustrated in F were characteristic ofthe rest of the tectum.
	Fig. 1. Distribution of galanin-like immunoreactivity in the brain of Rana esculenta. The drawing below E shows the transverse planes chosen for the illustrations. A, B, and C are from the telencephalon, whileD and E are from the diencephalon. The pre tectum is represented by F, while the mesencephalon is represented by G and H. I, J, K, and L are from the rhombencephalon. The location of immunoreactive nerve fibers, terminals, and perikarya is represented in the left half of each drawing, while the neuroanatomical parcellations are designated in the right half. The darkest pattern indicates the sites where the most intense immunoreactivity was detected. Dense patterns of small dots indicate areas where a considerable number of labelled axon terminals were detected. Crosses represent immunoreactive fibers and scattered terminals. Filled triangles represent intensely stained perikarya, while empty triangles indicate faintly stained cells. The three layers of immunoreactive nerve terminals in the optic tectum illustrated in F were characteristic ofthe rest of the tectum.
	Fig. 1. Distribution of galanin-like immunoreactivity in the brain of Rana esculenta. The drawing below E shows the transverse planes chosen for the illustrations. A, B, and C are from the telencephalon, whileD and E are from the diencephalon. The pre tectum is represented by F, while the mesencephalon is represented by G and H. I, J, K, and L are from the rhombencephalon. The location of immunoreactive nerve fibers, terminals, and perikarya is represented in the left half of each drawing, while the neuroanatomical parcellations are designated in the right half. The darkest pattern indicates the sites where the most intense immunoreactivity was detected. Dense patterns of small dots indicate areas where a considerable number of labelled axon terminals were detected. Crosses represent immunoreactive fibers and scattered terminals. Filled triangles represent intensely stained perikarya, while empty triangles indicate faintly stained cells. The three layers of immunoreactive nerve terminals in the optic tectum illustrated in F were characteristic ofthe rest of the tectum.
	Fig. 2. Galanin-like immunoreactivity in the medial amygdala. Immunoreactive perikarya are embedded in a dense network of immunoreactive fibers, most of which are probably terminals. Transverse section. Silver-DAB method. Scale bar: 50 um.
	Fig. 3. Galanin-like immunoreactivity in the preoptic area. Immunoreactive neurons form 3-4 rows around the preoptic recess, while immunostained fibers occupy the lateral part of the area. Transverse section. Silver-DAB method. Scale bar: 100 um.
	Fig. 4. Galanin-like immunoreactivity in the telencephalon at the level of the anterior commissure (a), rostral part of the preoptic recess of! and Just behind the accessory olfactory bulb (c). Note the abscence p nnmunoreactivity in the dorsal and lateral pallium and the greater part of the medial pallium. The area indicated by an arrow in (c) is shown under higher power in Figure 5. Transverse section. Silver-DAB method. Scale bar: 500 um.
	Fig. 5. Galanin-like immunoreactivity in the rostral part of the telencephalon. Note the dense accumulation of terminals in the periventriCular region and immunopositive fibers in Riithig's prominentia !ateralis. Transverse sections. Silver-DAB method. Scale bar is 50 um in each figure of the plate.
	Fig. 6. Galanin-like immunoreactivity in the lateral forebrain bundle at the level of the lateral amygdala. The thickest fiber-like structures may be dendrites of preoptic cells; however, it was not possible to find connections between perikarya and these structures. Transverse section. Nickel-DAB method.
	Fig. 7. Galanin-like immunoreactivity in the suprachiasmatic nucleus. Note the Golgi-like staining of neurons. Transverse section. Silver- DAB method. Asterisk labels the third ventricle.
	Fig. 8. Galanin-like immunoreactivity in the infundibulum. Arrows point axons of immunoreactive neurons, while open arrow points to median eminence. Transverse section. Nickel-DAB method.
	Fig. 9. Galanin-like immunoreactivity in the periventricular part of the infundibulum. Arrowheads point to immunoreactive cell processes which may contact the cerebrospinal fluid. Parasagittal section. Rostral is Upward; dorsal is to the right. Nickel-DAB method. Scale bar: 25 um.
	Fig. 10 Galanin-like immunoreactivity in the subependymal layer of the infundibular recess. The three cells indicated by arrows send their processes. parallel with the wall of the infundibular recess. Transverse section. Nickel-DAB method. Scale bar: 25 um.
	Fig.11. Galanin-like immunoreactivity in the rostral part of the diencephalon. Note the dense plexus of immunoreactive fibers around the neuropil of Bellonci. Transverse section. Silver-DAB method. Scale bar: 250 um.
	Fig. 12. Galanin-like immunoreactivity in the suprachiasmatic region (a) and the caudal one third of the diencephalon (b). Note the absence of immunoreactivity in the lateral forebrain bundle, the optic chiasm and the telencephalon (a), and the larger part of the thalamus and the optic tract (b). In (b) the arrow points to the dorsal fiber bundle which can be followed to the optic tectum. Star indicates the infundibulum. Transverse section. Silver-DAB method. Scale bar: 500 um.
	F!g. 13. Ultrastructural distribution of galanin-like immunoreactivIty m the infundibulum (a, b) and the deep layers of the optic tectum (c,d,e) of Rana esculenta. a: Galanin-containing neuron (arrows) located in the vicinity of two nonlabelled infundibular cells (Nand N1). The cytoplasm of the neuron contains labelled neurosecretory granules (arrowheads). Nu, nucleus. x 8,000. b: Galanin-containing axon (arrows) is wedged between two neurons (N and N1). The labelled fiber makes an rum-somatic synapse (arrowheads) with neuron N. x 38,000. c: Galanin-immunoreactive axon (large arrows) in the optic tectum exhibits labelled dense-core (arrowheads) and electron-lucent vesicles. A nonlabelled axon (A) terminates (small arrows) on a dendritic process (D). x 19,300. d and e: Galanin immunoreactive axons (arrows) make synaptic contacts (arrowheads) with dendrites (D) within layer 4 of the optic tectum. x 23,700 and x 30,400.
	Fig. 14. Galanin-like immunoreactivity in the rostral part of the tegmentum mesencephali. In addition to fibers, the perikarya of several neurons were stained. The periaqueductal plexus of immunoreactive fibers continues ventrally, to the surface of the brain. Transverse section. Silver-DAB method. Scale bar: 100um.
	Fig. 15. Galanin-like immunoreactivity in the rostral part of the mesencephalon. The dorsal immunoreactive fiber bundle (arrow in a) splits mto a ventral branch (open arrow) which intermingles with the periaqueductal plexus and a lateral branch which is continuous with lamina G (arrows in b) of the optic tectum. Transverse section. Silver-DAB method. Scale bar: 250 um.
	Fig. 16. Galanin-like immunoreactivity in the middle one third of the mesencephalon. Note the intense immunoreactivity in the interpeduncular nucleus (arrow in the left lower corner) and the mesenencephalic tegmentum, except the nucleus profundus and part of the torus semicircularis where only scattered immunoreactive fibers occurred. transverse section. Silver-DAB method. Scale bar: 500 um.
	Fig. 17. Laminar distribution of galanin-like immunoreactivity in the optic tectum. In the superficial layers (a), three immunoreactive bands can be distinguished (laminae B, D, and G+ part of layer 8). In the deep layers (b), layers 3 and 5 contain numerous immunoreactive fibers, while only scattered fibers occur in layer 6. Capital letters indicate tectal laminae; arabic numerals label tecta! layers. Transverse section. Silver-DAB method. Scale bar: 50 um.
	Fig. 18. Galanin-like immunoreactivity in the caudal-most part of the mesencephalon. Note the intense immunostaining in the isthmic nucleus. Transverse section. Silver-DAB method. The scale bar is 250 um in each figure of the plate.
	Fig. 19. Galanin-like immunoreactivity in the obex region. lmmunostained cells are located in the nucleus of the solitary tract. 'rhe 4th ventricle is indicated by an asterisk. Transverse section. Silver-DAB method.
	Fig. 20. Galanin-like immunoreactivity in the closed part of the medulla oblongata. Only fibers are stained. Transverse section. Silver DAB method.
	Fig. 21. Galanin-like immunoreactivity in the nucleus of the solitary tract and the dorsolateral fiber bundle. Horizontal section. Silver-DAB method.
	Fig. 22. Galanin-like immunoreactivity in the medulla oblongata. Note the heavily labelled fiber bundles. Nickel-DAB method. Sagittal section.
	Fig. 23. Galanin-like immunoreactivity in the preoptic area. Galanin containing cell bodies in the medial periventricular portion of the preoptic area and immunoreactive fibers in the lateral part of the preoptic area (white asterisk). The 3rd ventricle is indicated by a solid black asterisk. Xenopus laevis. Transverse section. Scale bar: 200 um.
	Fig. 24. Galanin-immunoreactive neurons in the periventricular part of the infundibulum. Some of the processes ofthe immunolabelled neurons appear to penetrate the wall of the 3rd ventricle (arrowheads). The main process of the immunoreactive cells runs lateroventrally. The 3rd ventricle is indicated by asterisks. Xenopus laevis. Transverse section. Scale bar: 40 um.
	Fig. 25. Galanin-immunoreactive fibers and nerve terminals in the central gray of the medulla. The asterisk indicates the ependymal wall of the 4th ventricle. Xenopus laevis. Transverse section. Scale bar: 200 um.
	Fig. 26. Galanin immunoreactive nerve fibers in the cerebellar nucleus (open star) and below the granule cell layer of the cerebellum (arrows). The 4th ventricle is indicated by an asterisk. Xenopus laevis. Transverse section. Scale bar: 200 um.
	Fig. 27. High-power photomicrograph of the optic tectum. Galanin immunoreactive nerve fibers are scattered throughout the tectum without any obvious laminar organization. The optic ventricle is indicated by asterisks. Xenopus laeuis. Transverse section. Scale bar: 40 um.
	Fig. 28. Galanin immunoreactive fibers in the nucleus of the solitary tract. The fourth ventricle is indicated by asterisks. Xenopus laevis. Transverse section. Scale bar: 200 um.
	Fig. 29. Galanin immunoreactive fibers around the central canal (asterisk) of the spinal cord. Xenopus laeuis. Transverse section. Scale bar: 200 um.