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Introduction: The distribution of the immunoreactive cell bodies and fibers containing neurotensin in the alpaca diencephalon was determined by an immunohistochemical technique., Material and Methods: The study was carried out in four male alpacas that lived at sea level. Brains of deeply anesthetized animals were fixed by perfusion with 4% paraformaldehyde. Cryostat sections were stained by a standard immunohistochemical method., Results: Cell bodies containing neurotensin were observed in the zona incerta and hypothalamus. A low/moderate density of these cell bodies was observed in the lateral hypothalamic area, anterior and dorsal hypothalamic areas, suprachiasmatic nucleus, periventricular region of the hypothalamus and in the ventromedial hypothalamic nucleus. In both thalamus and hypothalamus, immunoreactive fibers showed a widespread distribution. In the thalamus, a high density of these fibers was mainly found in the midline nuclei, whereas in the hypothalamus a high density was in general observed in the whole structure., Conclusions: In comparison with other mammals, the thalamus of the alpaca showed the most widespread distribution of neurotensin-immunoreactive fibers. The widespread distribution of neurotensin through the alpaca diencephalon suggests that the peptide can be involved in many physiological actions.

Using an immunohistochemical technique, we mapped the immunoreactive structures containing methionine-enkephalin-Arg 6 -Gly 7 -Leu 8 (Met-8) (a marker for the pro-enkephalin system) in the human diencephalon. Compared with previous studies, we observed a more widespread distribution of Met-8 in the human diencephalon. Met-8-immunoreactive cell bodies and fibers exhibited a more widespread distribution in the hypothalamus than in the thalamus. We observed six populations of Met-8-immunoreactive cell bodies. These perikarya were observed in the paratenial thalamic nucleus, ventromedial and dorsomedial hypothalamic nuclei, lateral hypothalamic area, pallidohypothalamic nucleus and in the paraventricular hypothalamic nucleus (posterior part). In the thalamus, Met-8-immunoreactive fibers were primarily observed in the midline region, whereas in the hypothalamus, these fibers were widely distributed. In general, a moderate/low density of Met-8-immunoreactive fibers was observed in the diencephalic nuclei. A moderate density was observed in the paraventricular thalamic nucleus, reuniens thalamic nucleus, lateral and medial geniculate nuclei, dorsomedial hypothalamic nucleus, paraventricular hypothalamic nucleus (posterior part) and ventromedial hypothalamic nucleus. The present study is the first to demonstrate the presence of clusters of Met-8-immunoreactive cell bodies in the human thalamus and hypothalamus, the distribution of fibers containing neuropeptides in the hypothalamus and the presence of these fibers in several thalamic nuclei. This neuroanatomical study will serve to elucidate the physiological roles of Met-8 in future studies of the human diencephalon., (Copyright © 2016 IBRO. Published by Elsevier Ltd. All rights reserved.)

Using an indirect immunoperoxidase technique, we studied the distribution of cell bodies and fibres containing non-opioid peptides (adrenocorticotropin hormone (ACTH), alpha-melanocyte-stimulating hormone) and opioid peptides (beta-endorphin (1-27), alpha-neo-endorphin, leucine-enkephalin) in the alpaca diencephalon. No immunoreactive cell bodies containing ACTH were found. Perikarya containing the other four peptides were observed exclusively in the hypothalamus and their distribution was restricted. Perikarya containing alpha-melanocyte-stimulating hormone or alpha-neo-endorphin showed a more widespread distribution than those containing leucine-enkephalin or beta-endorphin (1-27). Cell bodies containing pro-opiomelanocortin-derived peptides were observed in the arcuate nucleus, anterior and lateral hypothalamic areas and in the ventromedial and supraoptic hypothalamic nuclei, whereas perikarya containing alpha-neo-endorphin (a pro-dynorphin-derived peptide) were found in the arcuate nucleus, dorsal and lateral hypothalamic areas, and in the paraventricular, ventromedial and supraoptic hypothalamic nuclei. Immunoreactive cell bodies containing leucine-enkephalin were found in the lateral hypothalamic area and in the paraventricular hypothalamic nucleus. Immunoreactive fibres expressing pro-opiomelanocortin-derived peptides were more numerous than those expressing pro-dynorphin-derived peptides. A close anatomical relationship was observed: in all the diencephalic nuclei in which beta-endorphin (1-27)-immunoreactive fibres were found, fibres containing alpha-melanocyte-stimulating hormone or alpha-neo-endorphin were also observed. Fibres containing beta-endorphin (1-27), alpha-melanocyte-stimulating hormone or alpha-neo-endorphin were widely distributed throughout the diencephalon, but fibres containing ACTH or leucine-enkephalin showed a moderate distribution. The distribution of the five peptides studied here is also compared with that reported previously in other mammalian species. The widespread distribution observed indicates that both the pro-dynorphin and the pro-opiomelanocortin systems are involved in multiple physiological actions (e.g., food intake, thermoregulation, neuroendocrine and reproductive mechanisms) in the alpaca diencephalon., (Copyright © 2014 Elsevier B.V. All rights reserved.)

We report the distribution of immunoreactive cell bodies and fibers containing calcitonin gene-related peptide in the alpaca diencephalon. This study was carried out in alpacas that lived from birth to death at 0 m above sea level. Immunoreactive fibers were widely distributed throughout the thalamus and hypothalamus. A moderate density of these fibers was found in the zona incerta, the central medial, subparafascicular, reuniens and rhomboid thalamic nuclei, in the preoptic, anterior, lateral and dorsal hypothalamic areas, around the fornix, in the posterior, ventromedial and paraventricular hypothalamic nuclei and in the lateral mammillary nucleus. Cell bodies were only found in the hypothalamus: a high density in the paraventricular and supraoptic hypothalamic nuclei and a low density in the anterior, lateral and dorsal hypothalamic areas, around the fornix, and in the posterior and ventromedial hypothalamic nuclei. The widespread distribution of calcitonin gene-related peptide in the alpaca diencephalon suggests that it is involved in many physiological actions that must be investigated in-depth in the future, since alpacas lives from 0 m above sea level to altitudes of up to 5000 m altitude and hence the involvement of neuropeptides in special and unique regulatory physiological mechanisms could be suggested., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Using an immunocytochemical technique, we report for the first time the distribution of immunoreactive cell bodies and fibers containing somatostatin-28 (1-12) in the alpaca diencephalon. Somatostatin-28 (1-12)-immunoreactive cell bodies were only observed in the hypothalamus (lateral hypothalamic area, arcuate nucleus and ventromedial hypothalamic nucleus). However, immunoreactive fibers were widely distributed throughout the thalamus and hypothalamus. A high density of such fibers was observed in the central medial thalamic nucleus, laterodorsal thalamic nucleus, lateral habenular nucleus, mediodorsal thalamic nucleus, paraventricular thalamic nucleus, reuniens thalamic nucleus, rhomboid thalamic nucleus, subparafascicular thalamic nucleus, anterior hypothalamic area, arcuate nucleus, dorsal hypothalamic area, around the fornix, lateral hypothalamic area, lateral mammilary nucleus, posterior hypothalamic nucleus, paraventricular hypothalamic nucleus, suprachiasmatic nucleus, supraoptic hypothalamic nucleus, and in the ventromedial hypothalamic nucleus. The widespread distribution of somatostatin-28 (1-12) in the thalamus and hypothalamus of the alpaca suggests that the neuropeptide could be involved in many physiological actions., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Systemic subchronic oxytocin treatment significantly and substantially increased the B(max) values of the alpha 2 agonist [(3)H]UK14.304 binding sites in the hypothalamus, the amygdala and the paraventricular thalamic nucleus of the rat as shown by quantitative receptor autoradiography. These results suggest that long-term modulation of autonomic and neuroendocrine functions and emotional behaviours elicited by brain oxytocin may involve enhancement of central alpha 2-adrenoceptor function.

The distribution of luteinizing hormone-releasing hormone (LH-RH)-immunostained cell bodies and fibres was studied in the brainstem and diencephalon of the cat using an indirect immunoperoxidase technique. The brainstem and the thalamus were devoid of immunostained cell bodies, whereas in the hypothalamus immunopositive perikarya were observed in the supraoptic nucleus, the anterior hypothalamus, the preoptic region and in the arcuate nucleus. Our findings also showed that the hypothalamus is richer in immunostained fibres, and that in this region such fibres are more widely distributed than in the thalamus and upper brainstem. No immunopositive fibres were observed in the lower brainstem. Our results point to a more widespread distribution of LH-RH-immunostained perikarya in the cat hypothalamus than that previously reported in the cat; a similar distribution to that found in the rat, and a more restricted distribution than in primates. Additionally, our study shows a more widespread distribution of immunostained fibres in the cat brainstem and diencephalon than that previously described for other mammals. In this context, our results describe for the first time in the mammals central nervous system fibres containing LH-RH located in the stria medullaris of the thalamus, the supramammillary decussation, the laterodorsal and lateroposterior thalamic nuclei, the nucleus reuniens, the supraoptic nucleus, and the optic chiasm. Thus, our findings reveal that LH-RH-immunostained structures are widely distributed in the upper brainstem and in the diencephalon of the cat, suggesting that the peptide may be involved in several physiological functions.

Using an indirect immunoperoxidase technique, the location of cell bodies and fibres containing adrenocorticotropin hormone/corticotropin-like intermediate lobe peptide (ACTH/CLIP) was studied in the cat diencephalon. Immunoreactivity was observed in several diencephalic nuclei of the cat in which no immunoreactivity has been previously reported. In this sense, a low density of immunoreactive cell bodies was found in the nucleus ventromedialis hypothalami; a high density of immunoreactive fibres was found in the medial preoptic area; a moderate density in the lateral preoptic area and in the nuclei centralis thalami (pars medialis), interventralis thalami, interanteromedialis thalami, parafascicularis and praemamillaris (pars ventralis and pars dorsalis); a low density in the nuclei habenularis lateralis and reuniens thalami, and single fibres were found in the nuclei lateralis thalami (pars anterior), habenularis medialis, parataenialis, corpus geniculatum mediale, ventralis thalami (pars medialis) and in the fornix. Our results point to a more widespread distribution of ACTH/CLIP immunoreactive structures in the cat diencephalon in comparison with previous studies carried out in the same region of this feline.

The distribution of beta-endorphin (1-27) immunoreactive cell bodies and fibres was studied in the diencephalon of the cat using an indirect immunoperoxidase technique. In the thalamus, almost all the immunoreactive fibres were found in the midline region and in nuclei located near the midline, whereas in the hypothalamus fibres containing beta-endorphin (1-27) were visualized extending by the whole structure. The hypothalamus showed a higher density of beta-endorphin (1-27) immunoreactive fibres than the thalamus, as well as immunoreactive cell bodies, since in the thalamus no beta-endorphin (1-27) immunoreactive neuron was located. The densest network of immunoreactive fibres was observed in the epithalamus (nucleus periventricularis anterior) and in the hypothalamic nuclei arcuatus, hypothalami ventromedialis, suprachiasmaticus, periventricularis hypothalami, hypothalamus dorsomedialis, area hypothalamica dorsalis, hypothalamus anterior, filiformis, hypothalamus posterior and regio praeoptica. In the hypothalamus, a high density of perikarya containing beta-endorphin (1-27) was observed in the nucleus arcuatus and a low density in the nucleus hypothalami ventromedialis. The distribution of beta-endorphin (1-27) immunoreactive fibres and perikarya is compared with the location of other neuropeptides in the cat diencephalon. Our findings reveal that b-endorphin (1-27) immunoreactive structures are widely distributed in the cat diencephalon, suggesting that the peptide might be involved in several physiological functions.

The distribution of calbindin D-28k-immunoreactive fibers and cell bodies in the cat diencephalon has been analyzed by using the avidin-biotin immunoperoxidase technique. The thalamus showed a higher density of immunoreactive cell bodies than the hypothalamus. A high density of both immunoreactive perikarya and fibers was observed in the nuclei pulvinar, geniculatum mediale and laterale, lateralis posterior and dorsalis, habenularis lateralis and medialis, posterior, subparafascicularis, submedius, centralis medialis, medialis dorsalis, ventralis postero-medialis and postero-lateralis, reticularis, centralis lateralis, rhomboidens, paracentralis, ventralis lateralis, anterior and medialis, reuniens, anterior ventralis and medialis, hypothalamus posterior, corpus mamillare, area hypothalamica dorsalis and in the hypothalami ventromedialis. Moreover, a high density of fibers containing calbindin and a moderate/low density of immunoreactive cell bodies was found in the nuclei periventricularis anterior, parataenialis, hypothalamus lateralis, mamillaris lateralis, filiformis, periventricularis hypothalami, hypothalamus anterior and in the suprachiasmaticus.

The distribution of parvalbumin-immunoreactive cell bodies and fibers in the cat diencephalon has been analyzed by using the avidin-biotin immunoperoxidase technique. The thalamus showed a higher density of immunoreactive cell bodies than the hypothalamus. A high or moderate density of perikarya and a high density of fibers containing parvalbumin was observed in the nuclei lateralis posterior, lateralis dorsalis, pulvinar, corpus geniculatum laterale, reticularis, medialis dorsalis, centrum medianum, subparafascicularis, ventralis postero-medialis, ventralis postero-lateralis, habenularis medialis, parafascicularis, corpus geniculatum mediale, centralis lateralis, rhomboidens, reuniens, centralis medialis, ventralis medialis, ventralis lateralis, parataenialis, anterior ventralis, anterior medialis, ventralis anterior, hypothalamus posterior, corpus mamillare, area hypothalamica dorsalis, and in the nucleus suprachiasmaticus. Moreover, a high or moderate density of immunoreactive fibers and a low density of parvalbumin-immunoreactive cell bodies was observed in the nuclei periventricularis anterior, anterior dorsalis, habenularis lateralis, corpus geniculatum laterale (pars ventralis), periventricularis hypothalami, hypothalamus lateralis, hypothalamus anterior, and in the hypothalamus dorsomedialis.

The distribution of neurokinin A-like immunoreactive cell bodies and fibers in the diencephalon of the cat was studied using an indirect immunoperoxidase technique. A high or moderate density of immunoreactive neurons was observed in the nuclei habenularis lateralis, medialis dorsalis, parafascicularis, hypothalamus posterior, area hypothalamica dorsalis, hypothalamus lateralis, periventricularis hypothalami, above the corpus mamillare, and in the perifornical area, whereas scarce immunoreactive perikarya were visualized in the nuclei reuniens, hypothalami ventromedialis, hypothalamus dorsomedialis, and mamillaris lateralis. The highest density of fibers containing neurokinin A was found in the nuclei periventricularis anterior, rhomboidens, centralis medialis, periventricularis hypothalami, and supraopticus. In the regio praeoptica, area hypothalamica dorsalis, hypothalamus posterior, and in the perifornical area a moderate density of immunoreactive fibers was observed, whereas the nuclei habenularis lateralis, medialis dorsalis, mamillaris lateralis, parataenialis, reuniens, habenularis medialis, filiformis, hypothalamus dorsomedialis, hypothalami ventromedialis, arcuatus, and suprachiasmaticus showed a low density of neurokinin A immunoreactive fibers.

Using an indirect immunoperoxidase technique, the location of somatostatin-28 (1-12)-like immunoreactive fibres and cell bodies in the cat diencephalon was studied. The hypothalamus was richer in somatostatin-28 (1-12)-like immunoreactive structures than the thalamus. A high density of immunoreactive fibres was observed in the nuclei habenularis lateralis, paraventricularis anterior (its caudal part), filiformis, hypothalami ventromedialis, and regio praeoptica, whereas a moderate density was found in the nuclei paracentralis, supraopticus, supra chiasmaticus, hypothalamus posterior and area hypothalamica dorsalis. The nuclei lateralis dorsalis, lateralis posterior, medialis dorsalis, rhomboidens, centralis medialis, ventralis medialis, reuniens, anterior dorsalis, parataenialis, interanteromedialis, hypothalamus lateralis, hypothalamus dorsomedialis and arcuatus had the lowest density of immunoreactive fibres. In addition, a high or moderate density of somatostatin-28 (1-12)-like immunoreactive cell bodies was observed in the nuclei paraventricularis hypothalami, supraopticus, supra chiasmaticus, area hypothalamics dorsalis, subparafascicularis, hypothalamus posterior and hypothalamus anterior, whereas scarce immunoreactive perikarya were visualized in the nuclei lateralis dorsalis and parafascicularis. The distribution of somatostatin-28 (1-12)-like immunoreactive structures is compared with the location of other neuropeptides in the cat diencephalon.

Using an indirect immunoperoxidase technique, the location of neurotensin-like fibers and cell bodies was studied in the diencephalon of the cat. The findings showed that the hypothalamus is richer in neurotensin-like-immunoreactive structures than the thalamus, and that neurotensin-like-immunoreactive structures are more widely distributed in the hypothalamus than in the thalamus. A high density of immunoreactive fibers was observed in the hypothalamic regions, area hypothalamica dorsalis, hypothalamus posterior, nucleus (n.) filiformis and n. arcuatus, whereas a moderate density was found in the n. parafascicularis, n. paraventricularis anterior, hypothalamus lateralis, median eminence and n. paraventricularis hypothalami. Other diencephalic regions such as n. lateralis posterior, n. lateralis dorsalis, n. medialis dorsalis, n. habenularis lateralis, n. centrum medianum, n. rhomboidens, n. reuniens, hypothalamus anterior, n. supra chiasmaticus, hypothalamus ventromedialis, n. supraopticus and hypothalamus dorsomedialis had the lowest density of immunoreactive fibers. In addition, the densest clusters of neurotensin-like perikarya were found in the n. arcuatus, n. centralis medialis and hypothalamus posterior, whereas the n. medialis dorsalis, n. paraventricularis anterior, n. reuniens, hypothalamus lateralis and hypothalamus ventromedialis had the lowest density. In the n. lateralis dorsalis, n. supraopticus, area hypothalamica dorsalis and n. supra chiasmaticus the density of immunoreactive perikarya was moderate.