Vasoactive-intestinal-polypeptide (VIP)-like immunoreactive cells in the skull base of rats

Summary We investigated the distribution of vasoactive intestinal polypeptide (VIP)-like immunoreactive cell bodies in relation to the major cerebral and internal carotid arteries at the skull base in rats. Acetylcholinesterase (AChE) histochemistry was also applied to investigate the localization of this enzyme. VIP staining revealed a few positive cell bodies in nerves close to the internal carotid artery at the base of the skull as well as in the cerebral arterial wall. Ganglion-like cell bodies were detectable within the greater superficial petrosal (GSP) nerve. AChE activity was observed in VIP-like immunoreactive cell bodies along the whole of the GSP nerve. These cell bodies in the GSP nerve may give rise to at least some of the perivascular VIP-and AChE-containing nerves of the internal carotid arteries at the base of the skull.     

Electron-microscopic investigations of vasoactive intestinal peptide (VIP)-like immunoreactive terminal formations in the lateral septum of the pigeon

Vasoactive intestinal peptide (VIP)-like immunoreactive terminal fields were examined in the lateral septum of the pigeon by means of immunocytochemistry. According to light-microscopic observations, these projections originated from VIP-like immunoreactive cerebrospinal fluid (CSF)-contacting neurons, which are located in the ependymal layer of the lateral septum and form a part of the lateral septal organ. The processes of these cells gave rise to dense terminal-like structures in the lateral septum. Pre-embedding immuno-electron microscopy revealed that VIP-like immunoreactive axon terminals had synaptoid contacts with perikarya of small VIP-immunonegative neurons of the lateral septum, which were characterized by an invaginated nucleus, numerous mitochondria, a well-developed Golgi apparatus, endoplasmic reticulum and a small number of dense-core vesicles (about 100 nm in diameter). VIP-like immunoreactive axons were also seen in contact with immunonegative dendrites in the lateral septum. In both axosomatic and axodendritic connections, VIP-like immunoreactive presynaptic terminals contained large dense-core vesicles, clusters of small vesicles and mitochondria. These findings suggest that VIP-immunoreactive neurons of the lateral septal organ project to small, presumably peptidergic nerve cells of the lateral septum and that the VIP-like neuropeptide serves as a neuromodulator (-transmitter) in this area.     

Septal afferents to the area dentata terminate on vasoactive intestinal polypeptide (VIP)-like immunoreactive,non-pyramidal neurons

Summary Thermic lesions in the medial septum of adult rats result in dark degeneration of terminal boutons in the stratum moleculare and hilus of the area dentata. While most of the degenerating terminals are in synaptic contact with non-reactive cells, part of them end on dendrites of VIP-like immunoreactive neurons.     

Vasoactive intestinal polypeptide (VIP)-like immunoreactivity in salivary glands.

VIP-like immunoreactivity was found in rat, cat and human salivary glands at a concentration of 17.0 – 29.3 pmol/g of tissue. The immunoreactivity was localised by immunocytochemistry in beaded nerve fibres which occurred in association with secretory acini, ducts, blood vessels and larger nerves. In salivary glands VIP may act as a neurotransmitter or neuromodulator involved in the regulation of blood flow and in the composition and volume of saliva.     

A new surgical approach to the skull base in rats

A new procedure is described to surgically approach the rat skull base and visualize the spheno-occipital synchondrosis. This region is difficult to assess in small laboratory animals without incurring damage to midline structures. This method, however, uses a lateral approach through an anterior triangle of the neck bounded by the sternocleidomastoid, digastric (posterior belly) and omohyoid muscles. No structures are transected in this procedure, thus preserving the underlying anatomy. This oblique angle of approach allows clear visualization of the mid-basicranium while sparing retraction or surgical trauma to respiratory and digestive structures such as the larynx, trachea and esophagus.     

Neuropeptide Y (NPY)-like immunoreactive amacrine cells in retinas of frog and goldfish

Summary The distribution of neuropeptide Y (NPY)-like immunoreactivity in rat, rabbit, chick, frog and goldfish retinas was investigated by immunohistochemistry. Positive results were observed only in the frog and goldfish retinas. NPY immunoreactivity was associated with a small population of amacrine cell bodies in the inner nuclear layer and cell processes in the inner plexiform layer of both retinas. In the frog retina, three distinct layers containing immunoreactivity were observed in the inner plexiform layer. In contrast, the immunoreactivity in the same area of the goldfish retina was more or less separated into two layers. Convincing evidence could not be found for the co-existence of NPY-like material with other putative transmitter-like substances in the two retinas.Radioimmunoassay revealed the presence of small amounts of NPY-like immunoreactivity in the rabbit retina; the goldfish and frog retinas contained significantly more immunoreactive material. High performance liquid chromatography of the immunoreactive material in frog and goldfish retinas showed each retina containing different molecular forms of NPY-like proteins, neither of which resembled porcine NPY or PYY.The endogenous NPY-like material of the frog retina can be released by potassium depolarisation in a calciumdependent way. In view of all these data an NPY-like protein must now be considered a potential retinal transmitter.     

Vasoactive intestinal polypeptide (VIP)-like immunoreactivity in anuran intestine

Summary Immunocytochemical and radioimmunological techniques with region specific antisera have been used to identify a vasoactive intestinal polypeptide-like material in the anuran intestine. Seven species of Anura were investigated: Bombina bombina, Alytes obstetricans, Rana temporaria, Rana esculenta, Hyla arborea, Hyla crepitans and Bufo bufo.In five of the species (A. obstetricans, R. temporaria, H. arborea, H. crepitans and B. bufo) vasoactive intestinal polypeptide-like immunoreactive mucosal endocrine cells and nerve fibres in all layers of the gut wall, were detected by both immunofluorescence and peroxidase-antiperoxidase methods. In the other two species, R. esculenta and B. bombina, no mucosal endocrine cells were detected although the vasoactive intestinal polypeptide-immunoreactive nerve fibres were plentiful.Radioimmunoassay showed the presence of significant amounts of vasoactive intestinal polypeptide-immunoreactivity in intestinal extracts from all species. The highest quantities were present in those anurans with both immunostained cells and nerves. Gel permeation chromatography showed that most of the vasoactive intestinal polypeptide-like peptide eluted in a position identical to that of natural mammalian (porcine) vasoactive intestinal polypeptide.The results indicate that a vasoactive intestinal polypeptide-like peptide is well represented in the Anura and that it is immunologically very similar to the mammalian peptide.Part of this work was presented at the European Society of Comparative Endocrinology, 1979; see Buchan et al. 1980a     

Vasoactive intestinal peptide (VIP)-like immunoreactivity in the human sympathetic ganglia

Vasoactive intestinal peptide immunoreactive (VIP-IR) nerve fibres and terminals, neurons and small granule containing cells were observed in human lumbal sympathetic ganglia. Electron-microscopically VIP-IR was localized in the large dense-cored vesicles in nerve terminals and on the membranes of the Golgi complexes in the neurons. A small population of principal ganglion cells was surrounded by VIP-IR nerve terminals. Most of these neurons contained acetylcholinesterase (AChE) enzyme but were not tyrosine hydroxylase-immunoreactive (TH-IR). All VIP-IR ganglion cells and most of the nerve fibres contained AChE but not TH-IR. It appears that in human sympathetic ganglia VIP is localized in the cholinergic neurons and nerve fibres and that the VIP-IR nerve terminals innervate mainly the cholinergic subpopulation of the sympathetic neurons.     

Vasoactive intestinal peptide (VIP)-like immunoreactivity in the human sympathetic ganglia

Summary Vasoactive intestinal peptide immunoreactive (VIP-IR) nerve fibres and terminals, neurons and small granule containing cells were observed in human lumbal sympathetic ganglia. Electron-microscopically VIP-IR was localized in the large dense-cored vesicles in nerve terminals and on the membranes of the Golgi complexes in the neurons. A small population of principal ganglion cells was surrounded by VIP-IR nerve terminals. Most of these neurons contained acetycholinesterase (AChE) enzyme but were not tyrosine hydroxylase-immnoreactive (TH-IR). All VIP-IR ganglion cells and most of the nerve fibres contained AChE but not TH-IR. It appears that in human sympathetic ganglia VIP is localized in the cholingergic neurons and nerve fibres and that the VIP-IR nerve terminals innervate mainly the cholinergic subpopulation of the sympathetic neurons.     

VIP (Vasoactive Intestinal Polypeptide) — immunoreactive innervation of the portal vein

Summary Nerve fibres displaying immunoreactivity for vasoactive intestinal polypeptide (VIP) were found in the wall of the portal vein in cats, guinea pigs, rats and mice. In whole-mount preparations a sparse network of VIP fibres was seen in the vessel wall. Electrical field stimulation of the rat portal vein in vitro caused a significant release of VIP. The results suggest that VIP ergic nerve fibres play a role in the regulation of portal blood flow.     

Innervation of the pancreas by vasoactive intestinal polypeptide (VIP) immunoreactive nerves

The vasoactive intestinal polypeptide (VIP) has been shown to exert effects on endocrine and exocrine pancreatic secretion. Immunocytochemistry reveals that VIP immunoreactive nerves occur in the porcine, canine, feline and avian pancreas. In the pancreas of pig and cat VIP nerves are abundant around non-immunoreactive nerve cell bodies of the intrapancreatic ganglia but scarce in the islets and in the exocrine parenchyma. In the dog pancreas, however, the intrapancreatic ganglia contain strongly immunoreactive VIP nerve cell bodies which give off axons that seem to heavily innervate vessels as well as endocrine and exocrine cells. We suggest that in the pig and cat the pancreatic VIP nerves mainly affect the activity of a second type of intrapancreatic neuron, whose transmitter is unknown, whereas in the dog pancreas VIP nerves directly contact their putative effector structures.     

Cellular localization and ontogeny of immunoreactive Vasoactive intestinal polypeptide (VIP) in the chicken gut

Summary Vasoactive intestinal polypeptide (VIP)-immunoreactive nerves were abundant along the entire digestive tract of the chicken. In the proventriculus, gizzard and small intestine VIP nerves were numerous around glands and less numerous in the smooth muscle. Submucosal blood vessels were often encircled by VIP nerves. VIP nerves were also seen in the submucosal and myenteric plexus. In the large intestines the VIP innervation of the smooth muscle was more predominant, while there was a rather sparse supply of VIP nerves around the base of the crypts. This innervation pattern was a consistent finding with four different VIP antisera. VIP-immunoreactive cells, however, were demonstrated with only three of the antisera. They were found scattered in the epithelium of the proventriculus and small and large intestines. The failure of one of the antisera to demonstrate endocrine cells suggests that the VIP-immunoreactive material in these cells differs from that in nerves. Conceivably, the material present in nerves represents VIP, while that in endocrine cells represents cross-reacting peptides or other molecular forms of VIP.VIP nerves appeared comparatively early in embryonic development. They appeared in the upper part of the digestive tract at 13 days of incubation and in the colon a few days before hatching; at this stage, only smooth muscle received VIP nerves. The adult pattern of innervation was established about two to four weeks after hatching. VIP-immunoreactive endocrine cells appeared in the intestines a few days before hatching. The adult frequency of occurrence was established about one week after hatching.     

A quantitative histochemistry technique for measuring regional distribution of acetylcholinesterase in the brain using digital scanning densitometry

Studies of brain acetylcholinesterase (AChE) are traditionally based on biochemical assays, immunoreactivity, and histochemistry. Conventional histochemistry yields rich morphological data from tissue sections but yields quantitative results only with great difficulty. Several histochemical methods developed in recent years, including microdensitometry, microphotometry, and video-based histochemistry, are effective in quantitative and detailed study of AChE in tissue sections. However, they are usually time-consuming. As we report here, we adapted digital scanning densitometry to quantitate AChE histochemical staining in brain sections. The AChE and butyrylcholinesterase (BuChE), as measured by the method, were heterogeneously distributed throughout the brain, results that are consistent with those obtained by biochemical methods. The staining intensity is dependent on section thickness, substrate concentration, and reaction time. The cholinesterase inhibitor methyl paraoxon significantly decreased AChE staining intensity. Furthermore, data acquired from densitometry are similar to those obtained by video-based microscopy or by spectrophotometry. The advantage of the densitometric measurements compared to other quantitative histochemical methods is that it is very rapid while collecting data that are equivalent in quality. Because the digital scanning densitometers provide high quality and sensitive imaging, wide dynamic ranges, and convenient image analysis software, they are very useful tools in quantitative histochemistry.     

Distribution and synaptology of vasoactive intestinal polypeptide (VIP) immunoreactive structures in the rat periaqueductal grey

Light microscopic analysis of the rat midbrain periaqueductal grey (PAG) showed vasoactive intestinal polypeptide immunoreactive (VIP-ir) neurons localized at the lateral and ventral walls of the aqueduct. Some varicose VIP-ir elements were detected closely associated with the ependyma. While several VIP-ir elements were encountered immediately under the ependyma, in a few cases, VIP-ir cell bodies were seen on the luminal surface of the ependymal cells lining the aqueduct. Electron microscopy revealed that most of these cells possessed the characteristics of a local circuit neuron. All VIP-ir cells had indented nuclei. Two types were distinguished: one with rounded cell body receiving numerous axo-somatic synapses established by VIP-negative axons. The other cell type was fusiform and its surface was almost fully isolated from axonal contacts by a glial sheath. The VIP-ir processes were interconnected with other periaqueductal cells by a variety of synaptic contacts. VIP-ir axon terminals formed asymmetric synapses with immunonegative dendritic shafts often in glomerulus-like assemblies. The postsynaptic immunonegative dendrites were of the aspinous, beaded type. We suggest that VIP-ir cells and processes in the midbrain PAG establish connections between the longitudinal functional columns of this region. On the basis of their morphology, VIP-ir cells in the PAG appear to be excitatory, terminating on inhibitory interneurons. Thus, a VIP-stimulated inhibition may be instrumental in the coordination of responses evoked by the stimulation of PAG columns.     

Vasoactive intestinal peptide (VIP) cells in the pancrease and gastro-intestinal mucosa. An immunohistochemical and ultrastructural study.

Using antibodies against pure porcine VIP in immunoperoxidase and immunofluorescence tests, VIP-immunoreactive cells have been detected in the pancreas-especially in the islets-and gastrointestinal mucosa of the dog, guinea-pig and man. VIP immunoreactive cells were widely distributed in these tissues, never being numerous at any site. Some parallelism has been noted between such cells and ultrastructurally identified D1 cells fo the pancreas and gastrointestinal mucosa. The presence of VIP cells in normal pancreas may help explain the occurrence of pancreatic endocrine tumors producing VIP.     

The origin of (Kolmer's) epiplexus cells. A combined histomorphological and histochemical study.

The morphology and the histochemically proofed enzyme pattern of the (Kolmer's) epiplexus cells found at the telencephalic chorioid plexus of humans indicate that these cells are of monocytogenetic origin, that they are monocytes and monocytogenetic macrophages respectively.     

Distribution, histochemistry and ultrastructure of somatostatin-like immunoreactive cells in the gastroenteric tract of the cartilaginous fish Scyliorhinus stellaris (L.)

Summary Somatostatin-like immunoreactive cells of an open type have been identified in the digestive tract of the cartilaginous fishScyliorhinus stellaris (L.) by the use of immunocytochemical techniques. In the stomach these cells are numerous both in the corpus (neck zone and tubular glands) and in the pyloric portion (crypts). In the spiral valve, somatostatin-like cells are rare, situated in the intestinal epithelium and without any particular localization. Using semithin serial sections,somatostatin-like cells are found to be Davenport-negative and weakly positive towards the Grimelius silver reaction, and using the semithin and ultrathin technique have been identified at the ultrastructural level; their secretory granules appear electron dense, round or slightly polygonal, and with a limiting membrane tightly adherent to the core. The mean diameter varies from 250–300 nm.     

Localisation of pancreatic polypeptide (PP)-like immunoreactive material in neurones of the brain of the blowfly,Calliphora erythrocephala (Diptera)

Summary The brain of the blowfly, Calliphora erythrocephala, has been studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibodies to bovine pancreatic polypeptide (BPP). A number of immunoreactive neurones have been localised, some corresponding to neurones previously identified tentatively as neurosecretory. This finding is further evidence that biologically active peptides, previously considered to be vertebrate, also exist in invertebrates. It also supports the concept of their evolutionary origin in nervous tissue.