Vasoactive intestinal polypeptide immunoreactivity in the spinal cord of the guinea pig

Summary The distribution of vasoactive intestinal polypeptide-immunoreactive (VIP-IR) neurons in the lower medulla oblongata and the spinal cord has been analyzed in guinea pigs. This study includes results obtained by colchicine treatment and transection experiments. In the spinal cord, numerous VIP-IR varicosities were observed in the substantia gelatinosa of the columna dorsalis; some were also found in the substantia intermedia and the columna anterior. The spinal VIP-IR nerve fibers were mainly of intraspinal origin and oriented segmentally. VIP-IR nuclei in the spinal cord extended dorsally into corresponding regions of the caudal medulla oblongata, namely from the substantia intermedia medialis and lateralis into the vagus-solitarius complex and from the nucleus spinalis lateralis into the area of the nucleus reticularis lateralis. Additional VIP-IR perikarya were observed in the pars caudalis of the nucleus spinalis nervi trigemini. The VIP-IR nuclei within the caudal medulla oblongata probably form a continuous system with those localized within the spinal cord. They may be involved functionally in the modulation of cardiovascular and respiratory regulation in the guinea pig.Supported by the DFG, Carvas SFB 90     

Morphology and ultrastructure of the adrenal gland in Bactrian camels (Camelus bactrianus)

In the present study, we examined the morphological features of the adrenal gland in Bactrian camel by means of digital anatomy, light and electron microscopy. Our findings testified that the gland was divided into three parts, capsule, cortex and medulla from outside to inside as other mammals, and the cortex itself was further distinguished into four zones: zona glomerulosa, zona intermedia, zona fasciculate and zona reticularis. Notably, the zona intermedia could be seen clearly in the glands from females and castrated males, whereas it was not morphologically clear in male. There was a great deal of lipid droplets in the zona fasciculate, while it was fewer in the zona glomerulosa and zona reticularis. The cytoplasm of adrenocortical cell contained rich mitochondria and endoplasmic reticulum. The adrenal medulla was well-developed with two separations of external and internal zones. The most obvious histological property of adrenal medulla cells were that they contained a huge number of electron-dense granules enveloped by the membrane, and so medulla cells could be divided into norepinephrine cells and epinephrine cells. Moreover, the cortical cuffs were frequently present in adrenal gland. Results of this study provides a theoretical basis necessary for ongoing investigations on Bactrian camels and their good adaptability in arid and semi-arid circumstances.     

Distribution of the fibers of the descending respiratory pathway in the phrenic nucleus of the cat

A distribution of degenerated fibers in contralateral phrenic nucleus of cats with chronic hemisection of the spinal cord under medulla oblongata has been studied. These fibers approach the nucleus after the partial secondary crossing of the descending respiratory pathway on the level of cervical segments. Degenerated fibers follow the dendrites of phrenic motoneurons and approach the basal segments of dendrites but do not reach the cell bodies. Possible mode of control over phrenic nucleus activity is under discussion.     

Distribution of gastrin and CCK-like peptides in rat brain

Summary The distribution of gastrin and CCK-like peptides in the rat brain was studied by immunocytochemistry using an antiserum reacting equally well with both groups of peptides. Immunoreactive nerve cell bodies were detected in all cortical areas, in the hippocampus where they were particularly numerous, in the mesencephalic central gray and in the medulla oblongata. After colchicine treatment immunoreactive material appeared also in cell bodies of the magnocellular hypothalamic system. Immunoreactive nerve fibers were widely distributed in the brain. Particularly dense accumulations were seen in the hippocampus near the ventral surface of the brain, in the caudate nucleus, in the interpeduncular nucleus, the parabrachial nucleus, the dorsal part of the medulla oblongata and in the dorsal horn of the spinal cord. In the hypothalamus immunoreactive nerve fibers were observed in all nuclei, being most frequent in the ventromedial, dorsal and lateral hypothalamic nuclei. A rich supply of nerve fibers was seen in the outer zone of the median eminence and in the neurohypophysis. From previous immunochemical analysis it appears that the peptide demonstrated in most parts of the brain is identical with CCK-8. In the neurosecretory cell bodies of the hypothalamus, the median eminence and the neurohypophysis, however, the immunoreactive material is probably identical with gastrin.     

Local application of somatostatin in the rat ventrolateral brain medulla induces apnea

Local injections of the tetradecapeptide somatostatin (SOM) into the brain stem region were performed in anesthetized and decerebrate rats. SOM administration (0.6-1.8 nmol) into the nucleus paragigantocellularis and the nucleus reticularis lateralis of the ventrolateral medulla oblongata induced ventilatory depression and apnea. The occurrence of apnea was dose dependent and attributed to the anesthetic depth, and it was seen within 60-240 s after injection. In anesthetized rats the apnea was seen as a termination or a continuous decrease in tidal volume while respiratory frequency remained unaltered. SOM-induced apnea was caused by depression of central inspiratory drive. SOM injections into the dorsal medulla were ineffective in eliciting apnea, although a ventilatory depression but no apnea was induced in the awake unanesthetized state. In addition to its effect on basal ventilation, SOM administration in the ventrolateral medulla resulted in a blunted ventilatory response to hypoxic and hypercapnic stimuli in anesthetized rats. We conclude that SOM has potent inhibitory effects on respiration that are specifically located in the nucleus paragigantocellularis and the nucleus reticularis lateralis.     

Immunohistochemical Localization of C-RFamide, a FMRF-related Peptide, in the Brain of the Goldfish, Carassius auratus

Carassius RFamide (C-RFa) is a novel peptide found in the brain of the Japanese crucian carp. It has been demonstrated that mRNA of C-RFa is present in the telencephalon, optic tectum, medulla oblongata, and proximal half of the eyeball in abundance. Immunohistochemical methods were employed to elucidate the distribution of the peptide in the brain of the goldfish (Carassius auratus) in detail. C-RFaimmunoreactive perikarya were observed in the olfactory bulb, the area ventralis telencephali pars dorsalis and lateralis, nucleus preopticus, nucleus preopticus periventricularis, nucleus lateralis tuberis pars posterioris, nucleus posterioris periventricularis, nucleus ventromedialis thalami, nucleus posterioris thalami, nucleus anterior tuberis, the oculomotor nucleus, nucleus reticularis superior and inferior, facial lobe, and vagal lobe. C-RFa immunoreactive fibers and nerve endings were present in the olfactory bulb, olfactory tract, area dorsalis telencephali pars centralis and medialis, area ventralis telencephali, midbrain tegmentum, diencephalon, medulla oblongata and pituitary. However, in the optic tectum the immunopositive perikarya and fibers were less abundant. Based on these results, some possible functions of C-RFa in the nervous system were discussed.     

Distribution of reticulospinal fibers and theri endings in the lumbar segments of the rat spinal cord]

After stereotaxis lexions in the nucleus reticularis gigantocellularis of the modulla oblongata and nucleus reticularis pontis caudalis, the distribution of degenerating nerve fibers in the lumbar segments of the spinal cord has been studied by silver impregnation methods of Nauta and Fink-Heimer. Degenerating reticulo-spinal fibers and fragments of axonal terminations were found in the area of n. motorius ventro-medialis and n. motorius ventro-lateralis, as well as partly in n. motorius dorso-lateralis close to motoneurons and their dendrites. Mainly they pass into layers VII and VIII. This fact indicates the existence of direct-reticulo-motoneuronal synaptic connections in rats, which coincides with electrophysiological data.     

Immunocytochemical localization of GABA containing neurons in the ventrolateral medulla oblongata of the rat

Localization of gamma-aminobutyric acid (GABA) in the ventrolateral medulla oblongata of the rat was studied, using antisera directed against GABA molecule fixed to bovine serum albumin. Within the rostral portion of the ventrolateral medulla, GABA-like immunoreactive neurons were found in the lateral wing of the raphe magnus and in the region of the paragigantocellular reticular nucleus. In the caudal portion of the ventrolateral medulla, a lesser number of GABA-stained neurons were found in the region around the nucleus reticularis lateralis. GABA-like immunoreactive punctate structures were also found throughout the ventrolateral medulla. These results provide further evidence for the existence of GABAergic neurons in the ventrolateral medulla oblongata of the rat.     

Peptide YY-like immunoreactivity in the central nervous system of the rat

The concentration of peptide YY (PYY)-like immunoreactivity in rat brain and spinal cord was determined by radioimmunoassay. The highest concentrations were found in the cervical spinal cord (18.1 +/- 1.3 ng/g, mean +/- S.E.M.) and in the medulla oblongata (16.3 +/- 1.5 ng/g). Lower amounts were found in the pons and in the hypothalamus. Chromatographic analysis of the PYY-like immunoreactivity from various regions of the brain revealed 95% of the immunoreactive material to be indistinguishable from synthetic porcine PYY. PYY-immunoreactive nerve cell bodies could be demonstrated by immunocytochemistry in the medulla oblongata of colchicine-treated rats, the largest group of cells being found in the midline area between and partly in the raphe pontis and obscurus nuclei. Another large group of immunoreactive cells was detected more laterally in the medial parts of the gigantocellular reticular nucleus. A few cells, finally, were seen in the dorsal parts of the medulla, including the nucleus of the solitary tract. Varicose nerve fibers displaying PYY immunoreactivity were observed in many parts of the hypothalamus, pons, medulla and spinal cord.     

The distribution pattern of alpha-MSH-like immunoreactivity in the cat central nervous system

The distribution pattern of alpha-melanocyte stimulating hormone-like immunoreactivity (alpha-MSH-Li) was studied in cats using avidin-biotin modification of immunocytochemical method. Cell bodies containing alpha-MSH-Li were observed in the medial basal hypothalamus, especially in the infundibular nucleus, the lateral hypothalamus and near zona incerta. Fibers with alpha-MSH-Li extended beyond the hypothalamus, into the paraventricular nucleus of the thalamus, rostral amygdala, periaqueductal gray, locus ceruleus, parabrachial nucleus and medial nucleus of the nucleus tractus solitarius. Axons with alpha-MSH-Li were also seen diffusely in various cortical areas, but more extensively in the limbic cortical regions. The distribution pattern of the cell bodies and fibers containing alpha-MSH-Li bears several similarities to that seen in rats, but differs in that the alpha-MSH-Li was not observed in cell bodies in locations other than the medial basal and lateral hypothalamus.     

Substance P in the ventrolateral medulla oblongata regulates ventilatory responses.

Local injection of substance P (SP) into the ventral portion of the nucleus gigantocellularis, nucleus reticularis lateralis, and nucleus retrofacialis of the ventrolateral medulla oblongata (VLM) or direct application on the ventral surface of the medulla oblongata caused marked stimulation of tidal volume (VT) and/or minute ventilation (VE). The ventilatory response to hypoxia was significantly blunted after SP in the VLM but not in the dorsal medulla oblongata (DM) (nucleus tractus solitarius). The SP antagonist [D-Pro2,D-Trp7,9]SP almost completely inhibited this response when applied locally to a wide area of the superficial layer of the VLM but not of the DM. Unilateral or bilateral application of 0.3-1.5 nmol of the SP antagonist in the VLM (corpus trapezoideum and the caudal region extending from the rootlets of the nucleus hypoglossus to the first cervical segment) markedly attenuated the response to a 5% CO2 inhalation. The inhibition of the CO2 response was seen after [D-Pro2,D-Trp7,9]SP in the rostral areas of the medulla oblongata corresponding to the corpus trapezoideum and the caudal region extending from the rootlets of the nucleus hypoglossus to the first cervical segment of the cervical cord. Electric somatosensory-induced ventilatory stimulation could be depressed by approximately 70% by [D-Pro2,D-Trp7,9]SP locally applied on the surface of the VLM. We conclude that SP is involved in the hypoxic, hypercapnic, and somatosensory ventilatory responses in the rat. However, these respiratory reflexes are mediated via different neuronal pools in the medulla oblongata, mainly the VLM.     

Distribution of NT-IR perikarya in the brain of the guinea pig with special reference to cardiovascular centers in the medulla oblongata

Summary The occurrence and distribution of neurotensin-immunoreactive (NT-IR) perikarya was studied in the central nervous system of the guinea pig using a newly raised antibody (KN 1). Numerous NT-IR perikarya were found in the nuclei amygdaloidei, nuclei septi interventriculare, hypothalamus, nucleus parafascicularis thalami, substantia grisea centralis mesencephali, ventral medulla oblongata, nucleus solitarius and spinal cord. The distribution of NT-IR perikarya was similar to that previously described in the rat and monkey. In the gyrus cinguli, hippocampus and nucleus olfactorius, though, no NT-IR neurons were detected in this investigation. Additional immunoreactive perikarya, however, were observed in areas of the ventral medulla oblongata, namely in the nucleus paragigantocellularis, nucleus retrofacialis and nucleus raphe obscurus.The relevance of the NT-IR perikarya within the ventral medulla oblongata is discussed with respect to other neuropeptides, which are found in this area, and to cardiovascular regulation.Abbreviations abl nucleus amygdaloideus basalis lateralis - abm nucleus amygdaloideus basalis medialis - acc nucleus amygdaloideus centralis - aco nucleus amygdaloideus corticalis - ahp area posterior hypothalami - ala nucleus amygdaloideus lateralis anterior - alp nucleus amygdaloideus lateralis posterior - ame nucleus amygdaloideus medialis - atv area tegmentalis ventralis - bst nucleus proprius striae terminalis - CA commissura anterior - CC corpus callosum - cgld corpus geniculatum laterale dorsale - cglv corpus geniculatum laterale ventrale - cgm corpus geniculatum mediale - CHO chiasma opticum - CI capsula interna - co nucleus commissuralis - cod nucleus cochlearis dorsalis - cp nucleus caudatus/Putamen - cs colliculus superior - cu nucleus cuneatus - dmh nucleus dorsomedialis hypothalami - DP decussatio pyramidum - em eminentia mediana - ent cortex entorhinalis - epi epiphysis - FLM fasciculus longitudinalis medialis - fm nucleus paraventricularis hypothalami pars filiformis - FX fornix - gd gyrus dentatus - gp globus pallidus - gr nucleus gracilis - hl nucleus habenulae lateralis - hm nucleus habenulae medialis - hpe hippocampus - ift nucleus infratrigeminalis - io oliva inferior - ip nucleus interpeduncularis - LM lemniscus medialis - MT tractus mamillo-thalamicus - na nucleus arcuatus - nls nucleus lateralis septi - nms nucleus medialis septi - npca nucleus proprius commissurae anterioris - ns nucleus solitarius - n III nucleus nervi oculomotorii - nt V nucleus tractus spinalis nervi trigemini - ntm nucleus mesencephalicus nervi trigemini - osc organum subcommissurale - P tractus cortico-spinalis - PC pedunculus cerebri - PCI pedunculus cerebellaris inferior - pir cortex piriformis - pol area praeoptica lateralis - pom area praeoptica medialis - prt area praetectalis - pt nucleus parataenialis - pvh nucleus paraventricularis hypothalami - pvt nucleus paraventricularis thalami - r nucleus ruber - re nucleus reuniens - rgi nucleus reticularis gigantocellularis - rl nucleus reticularis lateralis - rm nucleus raphe magnus - ro nucleus raphe obscurus - rp nucleus raphe pallidus - rpc nucleus reticularis parvocellularis - rpgc nucleus reticularis paragigantocellularis - sch nucleus suprachiasmaticus - SM stria medullaris thalami - snc substantia nigra compacta - snl substantia nigra lateralis - snr substantia nigra reticularis - ST stria terminalis - tad nucleus anterior dorsalis thalami - tam nucleus anterior medialis thalami - tav nucleus anterior ventralis thalami - tbl nucleus tuberolateralis - tc nucleus centralis thalami - tl nucleus lateralis thalami - tmd nucleus medialis dorsalis thalami - TO tractus opticus - TOL tractus olfactorium lateralis - tpo nucleus posterior thalami - tr nucleus reticularis thalami - trs nucleus triangularis septi - TS tractus solitarius - TS V tractus spinalis nervi trigemini - tvl nucleus ventrolateralis thalami - vmh nucleus ventromedialis hypothalami - vh ventral horn, Columna anterior - zi zona incerta Supported by the Deutsche Forschungsgesellschaft (DFG) SFB 90, Carvas     

Localization of GRF-like immunoreactive neurons in the rat brain

The localization of human GRF1-44-immunoreactive neurons was studied in the rat brain. A dense accumulation of GRF-containing fibers was noted in the external layer of the median eminence. Cell bodies were observed in colchicine-treated rats. The most intensely fluorescent cluster of cells was contained in the arcuate nucleus. Other cells were seen on the base of the hypothalamus, within the median forebrain bundle, dorsal and ventral aspects of the ventromedial nucleus, zona incerta and dorsal part of the dorsomedial nucleus. These cells may influence the pulsatile release of pituitary growth hormone.     

眼镜蛇毒对大鼠延髓一氧化氮合酶表达的影响

目的 探讨眼镜蛇毒对大鼠延髓某些核团一氧化氮合酶(NOS)表达的影响。方法 采用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)方法,观察大鼠延髓某些核团NOS阳性神经元在眼镜蛇毒中毒组、生理盐水组、正常对照组的变化。结果 蛇毒组大鼠延髓的中缝大核,外侧网状核NOS阳性神经元比对照组表达增强。结论 眼镜蛇毒对延髓的NOS阳性神经元表达有上调作用。     

Autoradiographic localization of vitamin A in the adrenal gland of rats.

Cellular distribution of vitamin A in the rat adrenal was evaluated by autoradiography. Vitamin A was concentrated in the lipid droplets of epithelial cells of the zona fasciculata and zona reticularis. A small amount of vitamin A was also present in the cytoplasm and nucleus of these cells. The zona glomerulosa contained very little vitamin A either in lipid droplets or in the remainder of the cell. The medulla had essentially no vitamin A. Such strong cellular specificity for uptake of vitamin A by the cells of the zona fasciculata and zona reticularis supports the hypothesis that vitamin A is involved in synthesis of glucocorticoid hormones.     

Immunocytochemical localization of adrenodoxin in bovine adrenal cortex by protein A-gold technique

To investigate the intracellular localization of the enzymes that are involved in steroid hormone synthesis, an immunocytochemical study of the distribution of adrenodoxin in cells of the bovine adrenal cortex was carried out by the post-embedding immunostaining method and by immuno-cryoultramicrotomy in combination with the protein A-gold technique. Gold particles were seen on the matrix and the inner membrane of all the mitochondria examined, which have typical vesicular or tubulo-vesicular cristae, in parenchymal cells of the zona fasciculata and the zona reticularis. Gold particles were distributed homogeneously among the mitochondria. The density of gold particles on mitochondria in the parenchymal cells of the zona glomerulosa was lower than that of the zona fasciculata, which was similar to that of the zona reticularis. Gold particles were also seen on round, electron-dense intramitochondrial bodies in the parenchymal cells. The intramitochondrial bodies were abundant in the zona glomerulosa and the outer region of the zona fasciculata.     

Galanin-like immunoreactivity in the chicken brain

The anatomical distribution of neurons containing galanin has been studied in the central nervous system of the chicken by means of immunocytochemistry using antisera against rat galanin. Major populations of immunostained perikarya were detected in several brain areas. The majority of galanin-immunoreactive cell bodies was present in the hypothalamus and in the caudal brainstem. Extensive groups of labeled perikarya were found in the paraventricular, periventricular, dorsomedial and tuberal hypothalamic nuclei, and in the nucleus of the solitary tract in the medulla oblongata. In the telencephalon, immunoreactive perikarya were observed in the preoptic area, in the lateral septal nucleus and in the hippocampus. The mesencephalon contained only a few galanin-positive perikarya located in the interpeduncular nucleus. Immunoreactive nerve fibers of varying density were detected in all subdivisions of the brain. Dense accumulations of galanin-positive fibers were seen in the preoptic area, periventricular region of the diencephalon, the ventral hypothalamus, the median eminence, the central gray of the brainstem, and the dorsomedial caudal medulla. The distributional pattern of galanin-immunoreactive neurons suggests a possible involvement of a galanin-like peptide in several neuroregulatory mechanisms.     

Immunocytochemical localization of adrenodoxin in bovine adrenal cortex by protein A-gold technique

Summary To investigate the intracellular localization of the enzymes that are involved in steroid hormone synthesis, an immunocytochemical study of the distribution of adrenodoxin in cells of the bovine adrenal cortex was carried out by the post-embedding immunostaining method and by immuno-cryoultramicrotomy in combination with the protein A-gold technique. Gold particles were seen on the matrix and the inner membrane of all the mitochondria examined, which have typical vesicular or tubulo-vesicular cristae, in parenchymal cells of the zona fasciculata and the zona reticularis. Gold particles were distributed homogeneously among the mitochondria. The density of gold particles on mitochondria in the parenchymal cells of the zona glomerulosa was lower than that of the zona fasciculata, which was similar to that of the zona reticularis. Gold particles were also seen on round, electron-dense intramitochondrial bodies in the parenchymal cells. The intramitochondrial bodies were abundant in the zona glomerulosa and the outer region of the zona fasciculata.Supported by a Grant-in-Aid for Scientific Research on Priority Areas (no. 63635505) from the Ministry of Education, Science and Culture of Japan     

Distribution of parvalbumin-immunoreactivity in the rat thalamus using a monoclonal antibody

1. The distribution of parvalbumin cell bodies and fibers in the thalamus of the rat was studied using a monoclonal antibody and the avidin-biotin-peroxidase method. The densest clusters of immunoreactive perikarya were observed in the nuclei ventralis posterior, reticularis, ventralis anterior and zona incerta, whereas the nuclei habenularis lateralis, lateralis posterior, lateralis, centralis lateralis and ventralis lateralis had the lowest density. In the nucleus geniculatum laterale ventralis, the density of parvalbumin cell bodies was intermediate. In all these thalamic nuclei, small, round or fusiform immunoreactive cells with short immunolabeled dendritic processes were observed. 2. The densest network of immunoreactive fibers was observed in the nuclei geniculatum laterale ventralis, reticularis and zona incerta. The nuclei geniculatum laterale dorsalis, ventralis posterior, medialis ventralis, ventralis anterior, anterior ventralis, anterior dorsalis and rhomboidens contained a moderate number of parvalbumin fibers, whereas the nuclei lateralis posterior, habenularis lateralis, parataenialis, centrum medianum, lateralis, centralis lateralis, ventralis lateralis, medialis dorsalis, anterior medialis, ventralis medialis and lateralis anterior had the lowest density of immunoreactive fibers. In addition, a large number of immunoreactive fibers was found in the lemniscus medialis and a scarce number in the stria medullaris. 3. No immunoreactive structure was observed in the nuclei habenularis medialis, paraventricularis, reuniens and geniculatum mediale. 4. Thus, perikarya and fibers containing parvalbumin are widely distributed throughout the thalamus of the rat, suggesting that parvalbumin might play a role, directly or indirectly, in limbic, visual and somatosensory mechanisms.     

Neurophysin pathways in the normal and hypophysectomized rat

Summary The hypothalamo-extrahypophyseal neurophysin pathways (HEH) and the three hypothalamic nuclei secreting neurophysins, the supraoptic (SON), paraventricular (PVN) and suprachiasmatic (SCN) nuclei, of normal and hypophysectomized rats were studied by application of the immunoperoxidase procedure. Eight well-defined HEH pathways were recognized. Their main sites of projection were: lateral septum and subfornical organ (1 and 2); tractus diagonalis (3); medial nucleus of the amygdala and lateral ventricle (4); nucleus periventricularis thalami, nucleus habenulae lateralis and periaqueductal gray (5); periaqueductal gray, pineal organ, collicular recess and subependymal region of the fourth ventricle (6); dorsomedial nucleus and premammillary area (7); perimammillary region, corpus trapezoideum, ventral surface of medulla oblongata, nucleus tractus solitarii, nucleus commissuralis, substantia gelatinosa and formatio reticularis lateralis of the medulla oblongata and spinal cord (8).Neurophysin fibers of unknown origin were found in the frontal cerebral cortex.It was noted that in pathway 5 the amount of immunostainable material undergoes changes with age.The three neurophysin-secreting nuclei reacted differently following hypophysectomy. Among the HEH pathways the only one that seemed to be affected by hypophysectomy was that innervating the lateral septum.It is suggested that the neurons that survive hypophysectomy either do not project to the neural lobe or, alternatively, display axon collaterals projecting outside the neural lobe. Such a neuronal population could be the origin of the HEH pathways.Supported by Grant RSM-80-13 from the Directión de Investigaciones, Universidad Austral de Chile.The authors wish to acknowledge the valuable help of Mrs. Elizabeth Santibá~nez and Mr. Genaro Alvial.