Localization and release of immunoreactive vasoactive intestinal polypeptide in bovine adrenal medulla

The concentration of immunoreactive (IR) vasoactive intestinal polypeptide (VIP) in extracts from bovine adrenal medulla was 29.9 +/- 7.2 pmol/g wet wt., which was about 100 times that of IR neurotensin and 30 times that of IR somatostatin. Chromatographic analysis showed that most of the IR-VIP was the same molecular size as synthetic VIP(1-28). On retrograde perfusion of isolated bovine adrenal gland, release of VIP with catecholamine (CA) was marked on stimulation with high K+, but slight on stimulation with acetylcholine, which induced marked release of CA. These results suggest that most of the VIP is localized not in CA storing granules in chromaffin cells, but in other intraadrenal neuronal components. In immunohistochemical studies, IR VIP fibers with large varicosities were observed around the vessels in the adrenal medulla.     

胎儿小肠生长抑素、胃泌素及血管活性肠肽免疫反应细胞的个体发生

目的：探讨生长抑素（SS）、胃泌素（Gas）、血管活性肠肽（VIP）免疫反应（IR）细胞在胎儿小肠中的个体发生及其分布。方法：免疫组织化学方法和图像分析技术。结果：SS-IR细胞和Gas-IR细胞于11周即可见于小肠三段绒毛上皮和尚未分化完全的肠腺细胞间。随着胎龄增长,SS-IR细胞数量和细胞内SS由少至多,其中以十二指肠细胞最多,回肠最少。Gas-IR细胞和细胞内的Gas在十二指肠中随胎龄增长逐渐增多;21周后空肠中则有所减少,回肠中则消失。VIP-IR细胞数量少,整个胎期细胞数量、形态及免疫染色强度均未见明显变化。结论：SS、Gas和VIP在胎儿小肠的内分泌细胞中表达,提示内分泌激素在胎儿小肠的发育过程中起调节作用。     

Comparison of chromogranins A, B, and secretogranin II in human adrenal medulla and pheochromocytoma

Human phenochromocytomas were analyzed for the presence of chromogranins A and B and secretogranin II (chromogranin C). In immunohistochemistry, a positive staining of the tumor cells was observed for all three antigens. For immunoblotting, a method had to be developed which prevented proteolytic degradation of the antigens. Boiling homogenates of freeze-dried tissues achieved this and in addition, led to a significant enrichment of these secretory proteins. All three antigens could be identified in the tumor by immunoblotting. Chromogranin A and B were the major components present in about equal amounts. The relative concentration of all these antigens in the tumors was similar to those in adrenal medulla. A method involving high pressure liquid chromatography is presented which allows the separation of the human chromogranins/secretogranins.     

Coexistence of vasoactive intestinal peptide and enkephalins in the adrenal chromaffin granules of the frog

The chromaffin cells of the frog adrenal gland were studied at the electron microscopic level by means of the immunoperoxidase technique. Using specific antibodies against Met-enkephalin, Leu-enkephalin and vasoactive intestinal peptide (VIP), the coexistence of the three neuropeptides in the chromaffin cells was demonstrated. Furthermore, all three peptides were co-located in the 200-400 nm dense-core vesicles which also stained for dopamine-beta-hydroxylase. These results, which suggest concomitant release of catecholamines and neuropeptides by the chromaffin cells, support the concept that VIP is involved in the stimulation of adrenal steroid secretion during stress conditions.     

肠血管活性多肽或生长抑素对大鼠肠淋巴细胞在肠淋巴组织分布的影响

目的：观察肠血管活性多肽（VIP）或生长抑素（SST）对大鼠肠淋巴细胞归巢至肠相关淋巴组织（GALT）的影响。方法：从肠系膜淋巴管插管引流淋巴液，淋巴细胞经VIP和SST体外孵育后，用^51Cr标记，将^51Cr-肠淋巴细胞朋股静脉回输入大鼠体内。取出各组织、器官，用γ计数器检测其放射性活度。结果：生理状况下，约10％^51Cr-肠淋巴细胞在短期内归巢至GALT。经VIP或SST孵育的^51Cr-肠淋巴细胞回输入大鼠体内后，在肠系膜淋巴结（1．85％，1．60％）用Peyer结（1.83%,1.56%)分布的百分比显著低于对照组（3．83％，3．85％），P＜0．05）。2种多肽对^51Cr-肠淋巴细胞在小肠弥散淋巴组织的分布无明显影响。结论：VIP或SST对大鼠肠淋巴细胞归巢至Peyer结和肠系膜淋巴结有一定的抑制作用。     

Identification of noradrenergic nerve terminals immunoreactive for neuropeptide Y and vasoactive intestinal peptide in the rat kidney

Cryostat-and vibratome-cut sections of rat kidneys were singly or doubly labeled to visualize immunoreactive tyrosine hydroxylase (THI), dopamine beta-hydroxylase (DBHI), vasoactive intestinal peptide (VIPI), and neuropeptide Y (NPYI). Rats were perfusion fixed with 2-4% paraformaldehyde with or without 0.15% picric acid and rinsed in buffer for 18-48 hr. Single antigens were labeled with horseradish per-oxidase in vibratome sections, whereas cryostat sections were used to label one antigen with perox-idase and another with a fluorophore in the same tissue section. A dense plexus of DBHI noradren-ergic nerves innervates the renal arterial tree, and such nerves innervate the interlobar veins and renal calyx as well. Immunoreactive NPY is colocal-ized in most of these nerves, but some intrarenal noradrenergic nerves do not contain NPY but do contain VIP immunoreactivity. The distribution of NPYI nerves resembles that of DBHI nerves, whereas most perivascular noradrenergic nerves immunoreactive for VIP innervate selected arcuate and interlobular arteries. A small population of nonadrenergic, VIPI nerves innervates the renal calyx.     

Release of immunoreactive vasoactive intestinal peptide (VIP) from the gallbladder in response to vagal stimulation

Immunoreactive VIP was detected in the gallbladder lumen and in the arterial blood and venous effluent from the gallbladder in fasting cats. During perfusion of the gallbladder in vivo there was a constant basal intraluminal secretion of VIP. The VIP concentration in the luminal effluent exceeded that in plasma supporting the notion that there was a release from the gallbladder tissue. The rate of secretion was significantly increased during efferent electrical stimulation of the peripheral cut end of the cervical vagal nerves, after blockade with atropine. A similar increase in concentration of VIP was seen in the venous effluent from the gallbladder. The results suggest a local release of VIP from intrinsic neurons within the gallbladder wall. This release is increased in response to activation of non-cholinergic fibres in the vagus nerves, suggesting a role for VIP in regulation of gallbladder functions.     

The distribution of neuropeptide Y-like immunoreactive neurons in the human medulla oblongata

We have described the distribution of neuropeptide Y-like immunoreactive neurons in the medulla oblongata of the adult human. The majority of neuropeptide Y-like immunoreactive cells were found in four regions of the medulla: the ventrolateral reticular formation, the dorsomedial medulla, the secondary sensory nuclei and the rostral raphe nuclei. The morphology of neuropeptide Y-like immunoreactive cells varied in each of these regions. In the ventrolateral reticular formation, the labelled neurons were round and pigmented caudal to the obex but elongated and non-pigmented rostral to the obex; in the dorsomedial medulla, they were triangular and pigmented caudal to but not rostral to the obex; in the secondary sensory nuclei, they were multipolar, non-pigmented and significantly smaller than in the other areas; in the rostral raphe nuclei, they were bipolar and non-pigmented. Colocalization studies revealed that many neuropeptide Y-like immunoreactive cells also synthesize monoamines, consistent with conclusions based on a quantitative comparison of their distributions. Neuropeptide Y-like immunoreactivity was present in about 25% of presumed noradrenaline-synthesizing cells in the caudal ventrolateral medulla (corresponding to the A1 region); about 50% of adrenaline- and 70% of presumed serotonin-synthesizing cells in the rostral ventrolateral medulla (C1 and B2-3 regions); 90-100% of presumed noradrenaline-synthesizing cells in the dorsomedial medulla at and above the obex (A2 region); about 50% of adrenaline-synthesizing cells in the rostral dorsomedial medulla (C2 region); about 5% of presumed serotonin-synthesizing cells in the rostral raphe nuclei (B2-3 region). The largest of these groups was the presumed serotonin-synthesizing cells that contained neuropeptide Y-like immunoreactivity in the rostral ventrolateral medulla. This is the first report of such a cell group in the medulla of any mammal, and emphasizes the neuroanatomical differences between humans and other species.     

The distribution of calcium,magnesium, copper and iron in the bovine adrenal medulla

The subcellular distributions of calcium, magnesium, catecholamine and protein have been measured in four batches of bovine adrenal medullae; in addition, copper and iron have been measured in the chromaffin granules. Calcium is mainly found in chromaffin granules; magnesium, by contrast, is largely cytoplasmic. The stoichiometry of calcium, magnesium and catecholamines in the chromaffin granule matrix is given and their intragranular concentrations are calculated using measurements of the internal water space of the granules. The chromaffin granules are found to behave as osmometers when the sucrose concentration of the supporting medium is varied between 0.3 and 0.7m; about 30% of the intragranular water is osmotically inactive. Copper is a component of the enzyme dopamine β-hydroxylase; approximately 40% of granule copper is present within the chromaffin granule matrix, the rest being membrane-bound. Iron, a component of cytochrome b₅₆₁, is retained in the membrane; there is twice as much iron as copper in the granule membrane.It is not known how high concentrations of catecholamines and ATP are maintained inside chromaffin granules (or synaptic vesicles). It is possible that metal ions are required to stabilise complexes of high molecular weight. The chromaffin granules are a major reservoir of calcium in adrenal medulla cells.     

The distribution of opioid binding subtypes in the bovine adrenal medulla

Autoradiography has been used to examine the distribution of opioid binding subtypes in the bovine adrenal gland. Specific opioid binding sites were restricted to the adrenal medulla. Kappa sites, labelled with [3H]bremazocine (in the presence of excess unlabelled mu and delta ligands), were highly concentrated over nerve tracts. These nerve tract associated binding sites were sensitive to competition by the endogenous opioid, dynorphin (1-13). Specific [3H]bremazocine binding sites were also found over the adrenal medullary chromaffin tissue. These binding sites were concentrated over the peripheral, adrenaline-containing region of the medulla and were sensitive to competition by diprenorphine but not dynorphin (1-13). Delta opioid sites, labelled with [3H][D-Ala2,D-Leu5] enkephalin (in the presence of excess unlabelled mu ligand) were selectively localized to the central, noradrenaline-containing region of the adrenal medulla. Mu opioid sites, labelled with [3H][D-Ala2, NMePhe4,Gly-ol5]enkephalin, were low in number and distributed throughout the adrenal medulla. These studies demonstrate that mu, delta and two distinct kappa opioid binding sites are differently distributed within the bovine adrenal medulla and suggest possible new sites of action for the adrenal medullary opioid peptides.     

S-100 protein in soft-tissue tumors derived from Schwann cells and melanocytes.

In soft tissues outside the central nervous system, S-100 protein is found normally only in Schwann cells. Using the peroxidase-antiperoxidase immunohistochemical method S-100 was also found in tumors derived from Schwann cells and melanocytes, including neurofibromas, neurilemomas, granular cell myoblastomas, cutaneous nevi, and malignant melanomas. S-100 was not detected in malignant Schwannomas, neuroblastomas, oat cell carcinomas, medullary carcinomas of the thyroid, paragangliomas, or meningiomas. S-100 was also absent from neoplasms of soft tissues not usually considered to arise from cells of neural crest origin. S-100 appears to be a useful marker for identifying neoplasms derived from Schwann cells and melanocytes.     

Protein S-100 in the histological diagnosis of tumors

Review deals with the use of the antibodies against protein S-100 in the diagnosis of tumours in man and experimental animals. Protein S-100 was first isolated from the bovine brain and then identified in glial and Schwann cells of the nervous system, epidermal Langerhans cells, myoepithelial cells of the salivary and mammary glands and in many other cells of various organs. In spite of the lack of specificity to any tissue antibodies against protein S-100 are widely used in the diagnosis of human tumours, mainly for the differentiation between neurogenic and soft tissue tumours (positive response to S-100 protein is regularly observed in benign schwannomas, neurofibromas and granular cell tumours while fibroblastic tumours are S100-negative), for the differentiation between nonpigmented melanomas (positive) and anaplastic carcinomas (negative). The knowledge of S-100 protein distribution in normal organ facilitates the establishment of the tumour origin. Antibodies against S-100 protein allowed one to establish the neurogenic origin of some rat soft tissue tumours which were considered otherwise to be of mesenchymal origin and gave additional proof of the neurogenic origin of the rat endocardial tumours.     

S-100 immunoreactive cells in the spleen and bursa of Fabricius of broiler chickens

The distribution of S-100 protein in spleens and bursae of Fabricius of broiler chickens with various diseases was investigated by an immunohistochemical method with antiserum to bovine S-100 protein. S-100 protein-positive cells were found in six cases of 34 chickens. In spleens, S-100 protein was detected in reticular cells in the ellipsoids of the sheathed arteries and eosinophilic granulocytes. Follicular dendritic cells in germinal centres were also S-100 protein-positive. In the bursa, reticular cells of the follicles and eosinophilic granulocytes in the follicular cortex and surface epithelium were positive for S-100 protein. The evidence suggests that S-100 protein may be involved in diseases of chickens.     

Regional distribution of guanine nucleotide-sensitive and guanine nucleotide-insensitive vasoactive intestinal peptide receptors in rat brain.

Based on the ability of guanine nucleotides to inhibit the binding of vasoactive intestinal peptide to its receptors, a guanosine 5'-triphosphate analog, guanylyl-imidodiphosphate, was used to differentiate two subtypes (or different functional states of a single subtype) of vasoactive intestinal peptide receptor in brain with in vitro autoradiography. In most brain regions, guanylyl-imidodiphosphate reduced vasoactive intestinal peptide binding between 40 and 60%. However, in the supraoptic nucleus, locus coeruleus, interpeduncular nucleus, facial nucleus, olfactory tubercle and periventricular hypothalamic nucleus, 80% or more of vasoactive intestinal peptide binding was inhibited. In other brain regions, including the medial geniculate, olfactory bulbs, and ventral thalamic nuclei, guanylyl-imidodiphosphate had little effect on vasoactive intestinal peptide binding. In liver, lung and intestine it also partly inhibited vasoactive intestinal peptide binding. Electrophoretic analysis of vasoactive intestinal peptide, covalently cross-linked to its receptors in brain membranes, revealed a pair of bands between 44,000 and 52,000 mol. wt, a component at 64,000 mol. wt and another at 92,000 mol. wt. All were displaceable with vasoactive intestinal peptide but guanylyl-imidodiphosphate displaced only the 64,000 mol. wt band suggesting that the GTP-sensitive vasoactive intestinal peptide receptor seen in brain sections has a molecular weight of about 61,000. The differential sensitivity to guanylyl-imidodiphosphate suggests the existence of at least two vasoactive intestinal peptide receptor subtypes in brain, with distinct regional distribution, and may reflect differential coupling to second messenger systems.     

An immunohistochemical study on the distribution of glial fibrillary acidic protein, S-100 protein, neuron-specific enolase, and neurofilament in medulloblastomas

In order to clarify the differentiation of medulloblastomas, the authors studied on the morphological features and immunohistochemical expression of glial fibrillary acidic protein (GFAP), S-100 protein, neuron-specific enolase (NSE), and neurofilament (NF) in 31 medulloblastomas. GFAP was detected only in a small number of tumor cells of 5 medulloblastomas; S-100 protein in both small tumor cells and some so-called spongioblastic cells in 16 medulloblastomas; NSE in the more abundant tumor cells and the matrix in 28 medulloblastomas; NF in a few tumor cells of 12 medulloblastomas; GFAP and NF in 2 medulloblastomas, but each of them in different tumor cells. These results suggest that medulloblastomas have a capacity of differentiation along neuronal and/or glial lines. The conventional morphological markers of differentiation in medulloblastomas such as spongioblastic cells and Homer Wright rosettes were not necessarily compatible with expression of immunohistochemical markers such as GFAP or NF. NSE and S-100 protein seem less valuable markers of differentiation because they were detected in both neuronal and glial elements. But NSE, which was observed in most medulloblastomas, might have a value as a marker for medulloblastomas.