Organization of the mossy fiber system of the rat studied in extended hippocampi. II. Experimental analysis of fiber distribution with silver impregnation methods.

The mossy fibers, a major intrinsic hippocampal pathway connecting the dentate granule cells with the pyramidal cells in CA4 and CA3, have been reexamined in rats using mainly Fink-Heimer silver impregnation methods for demonstration of degenerating axons. By extending isolated hippocampi and cutting sections normal to the long axis, simple two-dimensional reconstructions of both the lesions and the resultant degeneration could be made. In the hilus, the zone with the greatest concentration of degenerating boutons was found between the lesioned granule cells and the CA3 pyramidal cells abuting on the hilus; outside this zone the concentration declines rather rapidly. Degenerating boutons were also observed in low concentration up to 200-300 microgram septal and temporal to the lesion. The mossy fibers in CA3 nearest the hilus have an intrapyramidal course and display a lamellar organization with fibers from the granule cells of the medial blade lying deep to those from the dentate crest. These in turn lie deep to those from the graule cells of the lateral blade. A mediolateral difference in the projection of the graule cells on the CA3 pyramidal cells was discovered: fibers from the medial granule cells descend about 600 micrometer in the temporal direction, whereas fibers from the lateral granule cells descend about 1,200 micrometer. This causes a divergence of the fibers from one single level, especially of the part of the fibers, being farthest away from the hilus. The degree of descent of the fibers from each mediolateral position of the granule cells was constant at all septotemporal levels examined.

Changes with age in the morphology of the cochlear nerve in rats: light microscopy.

Cochlear nerve morphology was examined in a series of rats ranging in age from young adulthood to advanced age in order to assess the extent of fiber loss and the nature of degenerative changes with age. The animals were perfused via the aorta with mixed aldehydes. Blocks including the cochlear nerves were removed, embedded in Araldite, and sectioned in a plane transverse to the longitudinal axis of the nerve. Analysis of the material included counts of normal and degenerating fibers and of glial cells, maps of fiber packing densities, and measurements of the cross-sectional area of the nerve. The median number of normal fibers in the young adult animals (2-3 months) was 21,218. This number was reduced by 21% at 26.5 months and by 24% in the oldest group (35-36 months). The number of degenerating myelin sheaths was first seen to be significantly increased at 6 months, reached a peak at 26.5 months, and declined at 35-36 months. There was an age-related increase in the cross-sectional area of the nerve, amounting to about 60% at 26.5 months and to about 50% at 35-36 months. Fiber packing density decreased evenly with age over the area of the nerve. The increased cross-sectional area and decreased fiber packing density appeared to be related to increases in the thickness of myelin sheaths and in the area occupied by interneural elements.

Propriospinal fibers in the rat.

Segments of rat spinal cord were isolated by transecting in two places and sectioning all dorsal roots between the transections. Following this procedure, the areas of the gray and white matter are decreased by approximately 50% compared to normal. We feel, for reasons elaborated in the discussion, that the white matter of the isolated segments contains almost exclusively propriospinal axons. If this is accepted, then the axonal counts in this paper provide estimates of the numbers of propriospinal axons in rat spinal cord. In the isolated segments, the lateral funiculi contain 21,000 myelinated and 31,000 unmyelinated axons and the ventral funiculi 10,500 myelinated and 1,500 unmyelinated axons. The number of these fibers is approximately 33% of the number in unoperated spinal cords. Thus approximately one-third of the axons in rat sacral lateral and ventral funiculi are propriospinal, a lower figure than would have been predicted from classical work. The ratio of myelinated to unmyelinated fibers is higher for propriospinal fibers than for the other axons in these funiculi. Thus the propriospinal axons, as a group, are slightly larger than the other axons in these funiculi. This is against currently accepted thinking which generally regards the propriospinal fibers as the finest in the white matter of the cord. Finally, the quantification of propriospinal systems in these funiculi allows more precision in our thinking about the organization of the spinal cord.

Structure of the leech nerve cord: distribution of neurons and organization of fiber pathways.

The abdominal nerve cord of the leech Macrobdella decora was studied under the light and electron microscopes. The ganglionic cortex consists of six hemicone-shaped packets of neuronal perikarya and apical processes regularly assembled in bilaterally symmetric rows. The orderly projection of the apical processes into the hilum of the packets is also followed by an orderly distribution of their branches across the neuropile. This part of the ganglion is made of two symmetrical halves or hemineuropiles enclosing two types of nerve tissue: coarse and fine neuropiles. The coarse neuropile has seven longitudinal and four commissural tracts of fibers and a distinctively segregated synaptic zone. Nerve processes in this neuropile mostly proceed from the neurons in the ganglia and some are the branches of giant afferent axons. The fine neuropile includes several longitudinal tracts of fibers and a non-segregated synaptic zone. Most nerve processes in this neuropile are small afferent axons and some come from neurons in the ganglia. Bundles of axons in the connectives result by the orderly projection of the neuropile longitudinal tracts and together form fiber pathways connecting the synaptic zones of successive ganglia. Pathways of through-ganglia giant axons, linking the coarse neuropile synaptic zones, and of small axons, linking the fine neuropile synaptic zones, are described.

Central distribution of the efferent cells and the primary afferent fibers of the trigeminal nerve in Pleurodeles waltlii (Amphibia, Urodela).

As part of a study on the organization of the brainstem in a primitive group of vertebrates, the efferent cells and primary afferent fibers of the urodele amphibian Pleurodeles waltlii were examined by means of retrograde and anterograde axonal transport and anterograde degeneration. The trigeminal motor nucleus is located in the periventricular gray just medial to the sulcus limitans. Its rostral part is a band of pear-shaped cells lying parallel to the wall of the ventricle, whereas its caudal part is a round mass consisting of polygonal cells. In addition, a small group of scattered neurons is situated ventral to the rostral part of the nucleus. The primary afferent fibers enter the brainstem in the dorsal two-thirds of the trigeminal root. They diverge into a short ascending and a long descending tract. The former distributes its axons to the principal sensory trigeminal nucleus, which is an ill-defined cell group located at the ventrolateral edge of the periventricular gray. In the descending tract, the fibers of the ophthalmic nerve are predominantly located ventromedially, and those of the maxillomandibular nerve dorsolaterally. A fascicle of the ophthalmic nerve leaves the descending tract and, apparently, makes contact with the accessory abducens nucleus. The descending tract extends caudally into the three upper cervical segments of the spinal cord. The mesencephalic trigeminal nucleus consists of conspicuous large cells, which are scattered through the tectum of the mesencephalon. The cells with peripheral branches in the ophthalmic nerve are mainly located in the caudal half of the tectum, and those with peripheral branches in the maxillomandibular nerve in the rostral half. Collaterals of the central branches of the mesencephalic trigeminal system were traced to an area of the periventricular gray situated between the motor nucleus and the principal sensory nucleus of the trigeminus.

Distribution of neurotensin-like immunoreactivity in the diencephalon of the Japanese monkey (Macaca fuscata).

The distribution of neurotensin-like immunoreactive (NT-LI) neurons was examined in the thalamus and hypothalamus of the Japanese monkey (Macaca fuscata) by using the peroxidase-antiperoxidase immunocytochemistry technique. In the thalamus, NT-LI neuronal perikarya were distributed mainly in the midline nuclear group and the dorsomedial nucleus, and partially in the intralaminar nucleus: Immunoreactive fibers were mainly distributed in the midline nucleus, particularly in the nucleus rhomboidalis. Numerous immunoreactive fibers were also detected in the regions that contain the pathways to extrathalamic areas such as the stratum zonale and inferior thalamic peduncle. In the hypothalamus, many immunoreactive neuronal perikarya were distributed in the lateral hypothalamic area and in the arcuate nucleus. Immunoreactive fibers were disseminated throughout the hypothalamus, but they were dense in the preoptic area and sparse in the ventromedial nucleus. An accumulation of dense immunoreactive endings was also observed in the external layer of the median eminence. NT-LI fibers in the external layer of the median eminence were considered to represent nerve endings near portal vessels. Functional roles of neurotensin in the thalamus and hypothalamus are discussed from the anatomical point of view.

Visual system of the channel catfish (Ictalurus punctatus): III. Fiber order in the optic nerve and optic tract.

In channel catfish the ganglion cell axons leave the retina via a ring of approximately 13 separate optic papillae. Each papilla serves an area of retina extending from the central zone of the retina to the periphery. Papillae located at a dorsal position in the ring serve exclusively dorsal retina. Ventrally located papillae, however, have an exaggerated peripheral retinal representation, so that they serve mostly ventral retina but also some areas of peripheral retina dorsal to the nasal and temporal poles. The ganglion cell axon bundles departing from the retina via individual papillae were labelled with horseradish peroxidase, and sections of the optic pathway were examined to reveal the topographic organization of the fibers. The topographic order of the optic nerve was dissimilar to that of cichlids and goldfish. Fibers from individual papillae remained together throughout the optic nerve. Close to the optic nerve head, the papillae were arranged as a continuum around the U-shaped optic nerve, without the discontinuity in the representation of the ventral retina seen in other fish. Fibers associated with the dorsal papillae were located at the tip of the caudolateral arm of the U, and fibers from ventral papillae were on the rostromedial arm. Fibers from nasally and temporally located papillae were found on the base of the U. By the level of the optic chiasm the U shape had flattened out but retained the relative ordering of the papillae. Rotation of the nerve as it became the optic tract brought the representation of the ventral papillae to the dorsal pole of the tract, and the dorsal papillae to the ventral tract. It was only in the optic tract that rearrangement of fibers became apparent. As described above, the axons of some ganglion cells in dorsal, peripheral retina left the retina and travelled through the optic nerve with axons from extreme ventral retina. In the optic tract, these dorsal fibers joined the main body of fibers from the dorsal retina. The significance of these observations for theories of fiber rearrangement is discussed.

Area postrema of the goldfish, Carassius auratus: ultrastructure, fiber connections, and immunocytochemistry.

The area postrema in goldfish is a dorsal midline structure in the caudal medulla spanning the level of the obex. As in other vertebrates, the sinus capillaries of the area postrema in goldfish are fenestrated. In goldfish, however, the area postrema is organized in a unique laminar fashion; from superficial to deep: meninx, vasculature, palisade layer, cell body layer, and ventral neuropil layer. Virtually all of the neurons of the area postrema exhibit tyrosine hydroxylase-like immunoreactivity. Each immunoreactive neuron is essentially bipolar, with a short apical dendrite extending dorsally to reach the external basal lamina of the capillaries and a basal dendrite reaching into the subjacent layer of neuropil. The apical dendrites have no synaptic specializations and probably function as interoceptors detecting blood-borne chemicals that leak out of the fenestrated capillaries. The basal dendrites receive synaptic input both within the neuropil of the area postrema and in the commissural nucleus of Cajal into which they extend. Primary afferent fibers of the subdiaphragmatic branches of the vagus nerve terminate within the area postrema and commissural nucleus. Thus the neurons of the area postrema may serve not only as direct chemoreceptive interoceptors but may also receive input from other visceral afferent systems.

Normal nerve fibers in the barrel region of developing and adult mouse cortex.

The pattern of normal nerve fibers associated with barrel-shaped structures in the somatosensory cortex of both young and adult mice, has been studied using a reduced silver method. In adult animals, the "barrel" sides and septa can be seen to contain densely packed bundles of nerve fibers running vertically between layers III and V. In parasagittal sections, these fibers appear as very dark bands between adjacent barrels, while in tangenital sections the fibers, cut in cross-section, appear as rings of dark spots concentrated around the barrel edges. In contrast to this, barrels in immature animals have dark, evenly stained centers and pale, cell dense sides. This immature pattern can first be distinguished in 2-day-old animals and persists until 18 to 19 days. Between 18 and 24 days a change from the immature to the adult pattern occurs with the appearance of darkly stained, fine fibers within layer IV, particularly within the barrel sides. It is suggested that the immature pattern is due, primarily, to the staining of thalamic afferents while the adult pattern appears with the development of intracortical and association fibers. Electron microscopy, on tissue previously treated by the silver method, shows that the silver deposits are mainly attached to longitudinal elements of the axoplasm and not associated with myelin. This may explain the success of this method in showing fibers in young, unmyelinated brains.

Cytoarchitecture of the avian ventral lateral geniculate nucleus.

The avian thalamic ventral lateral geniculate nucleus (GLv) was studied by light microscopic techniques in order to understand its anatomy, neuronal composition, and the nature of its retinal and tectal afferents. The avian GLv is of considerable interest because physiological experiments show that it is the brain structure with the highest percentage of color-opponent responses (Maturana and Varela, '82). We used adult pigeons and quail for the present study. With Nissl techniques a predominance of medium-size neurons (58%) constitute the GLv. The shape, size, and orientation of the different neurons is highly variable throughout the GLv. With the Golgi methods, 5 classes of neurons are distinguished: I and IV (large), II (medium-size), III and V (small). Some class IV large neurons have bifurcated axons; no axons were distinguished on the small neurons. Optic fibers penetrating the GLv are often collateral branches of retinal axons that continue elsewhere. Fink-Heimer methods show that retinal axon terminals end around large and medium-size neurons and also reach the internal lamina of the GLv. HRP tracing shows that the large and medium-size neurons of the GLv project to the optic tectum. On the basis of comparisons between the cytoarchitecture of the GLv described here and the physiological findings previously reported (Maturana and Varela, '82; Pateromichelakis, '79), we suggest that: (1) large GLv neurons are the color-opponent units, (2) medium-size neurons are the movement-sensitive units, and (3) small neurons are either interneurons (local circuit neurons), or they might project to the area pretectalis or to some other GLv projection region not yet described.

The nervous system of the male Dinophilus gyrociliatus (Polychaeta, Dinophilidae): II. Electron microscopical reconstruction of nervous anatomy and effector cells.

All neuronal cells in the dwarf male of the dimorphic polychaete species Dinophilus gyrociliatus were individually identified by means of serial ultrathin sections. Altogether 68 neural cells--including 40 sensory neurons and 2 glial cells--constitute a small but complex nervous system. Fifty-three neural cells are located in three pairs of ganglia and connected by paired nerve cords. The prominent frontal ganglia, each consisting of a well-developed neuropile and surrounded by 20 or 21 neural cells, represent the animal's brain. The ventral ganglia contain only 2 neurons each. The penis ganglia--four cells each--are associated with the copulatory organ. A conspicuous circumpenial fiber mass surrounds the basal part of the penis. The effector cells--22 multiciliated epidermal cells, 34 muscle cells, and different gland cells (?)--were also reconstructed and their innervation was partly elucidated. Sensory-motor neurons were unambiguously identified. They are discussed in regard to the small body size of the animal. The male's nervous organization resembles a very simple rope ladder and may represent a reduced derivative of a nervous system in normal-sized males of monomorphic species. Similarities, however, also occur with the developing nervous system of a planktotrophic metatrochophore. The neuronal organization, with its two centers (frontal ganglia and ventral ganglia vs. penis ganglia and circumpenial fiber mass), accords well with the bipartite behavioral pattern, which is entirely devoted to locomotion and copulation, respectively.

Golgi and Nissl studies of the visual cortex of the bottlenose dolphin.

Nissl, Golgi and fibre preparations were made of the cerebral cortex of the lateral gyrus of the bottlenose dolphin (Tursiops truncatus) in the region where visual evoked potentials have been reported (Sokolov et al., '72; Ladygina et al., '78). In the adult the visual cortex is relatively thin (average about 1,300 micron) for so large a brain (fixed brain weight for a typical adult in our series was 1,330 g). Layers I, III, and VI are wide and represent three-quarters of the total cortical thickness. Layer I contains few cell bodies, while III and VI have a variety of pyramidal and nonpyramidal neurons. Layers II and V are narrow and contain striking palisades of darkly staining pyramidal cells that are particularly large in layer V. No clearly demarcated layer IV is present in the adult dolphin visual cortex. Many of the neurons identified with the Golgi technique are typical of pyramids in other mammals, with a single apical dendrite and a bouquet of basal dendrites, mostly highly spiny. Others are unusual in having bifurcated or oblique apical dendrites. Typical large and small spiny and nonspiny stellates are also found, mainly in layers III and VI. In addition various forms of spindle-shaped, bipolar and multipolar neurons are found in most layers. An 18-day-old brain shows signs of immaturity in its visual cortex. It is thinner (970 micron) and on average its neurons are smaller, paler, and more densely packed. Especially the pyramids of layer V are much smaller than in the adult. Also, a distinct "granular" band occurs between layers III and V and seems to be a rudimentary layer IV. At 3 years of age most of the adult features have developed, but layer IV is still detectable. No striking differences were observed in cell and fibre architecture between the cortex of the lateral gyrus and that of the so-called "calcarine" area that has also been considered as "visual." We concluded that, although different in many respects from other mammalian visual cortices, that of the dolphin is apparently well developed and differentiated.

Light microscopic localization of putative glycinergic neurons in the larval tiger salamander retina by immunocytochemical and autoradiographical methods.

Putative glycinergic neurons in the larval tiger salamander retina were localized by a comparative analysis of high affinity 3H-glycine uptake and glycine-like immunoreactivity (Gly-IR) at the light microscopic level. Commonly labeled neurons include at least three types of amacrine cell (Type IAd, Type IAb, Type IIAd; distinguished by soma location and dendritic ramification), cell bodies in the ganglion cell layer (GCL), and rarely observed Type II (inner) bipolar cells. With the increased resolution provided by Gly-IR, we identified a Type IAa amacrine cell, two types of Type IIAd amacrine cells, and Gly-IR interplexiform cells. Gly-IR axons in longitudinal sections of the optic nerve indicate the presence of Gly-IR ganglion cells. The percentage of labeled somas in the inner nuclear layer (INL) compared to all cells in each layer was similar for the two methods: 30-40% in INL 2 (middle layer of somas), 30-40% in INL 3 (inner layer of somas), and about 5% in the GCL. Labeled processes were found throughout the full thickness of the inner plexiform layer (IPL), but with a much denser band in the proximal half (sublamina b). The only major difference between the two methods (3H-glycine uptake vs. Gly-IR) was that Type I (outer) bipolar cells were labeled only by 3H-glycine uptake; these cells were more lightly labeled with silver grains than cell bodies in either INL 2 or INL 3. Postembed labeling of 1 micron Durcupan plastic sections for Gly-IR showed the same pattern, but with much higher resolution, as obtained with 10 micron cryostat sections. This study indicates extensive colocalization of labeling by both probes in INL 2, INL 3, the IPL, and the GCL. We conclude that Gly-IR can serve as a valid and reliable marker for glycine-containing neurons in this retina and suggest that glycine serves as a transmitter for several morphologically distinct types of amacrine cell, an interplexiform cell, and perhaps a small percentage of Type II bipolar cells and ganglion cells.

Corticotropin releasing factor-containing afferents to the lateral septum of the rat brain.

Corticotropin releasing factor (CRF)-containing afferents to the rat lateral septum (LS) have been determined by means of cobalt-enhanced immunohistochemistry, tracing of retrograde transport of horseradish peroxidase (HRP), and by lesioning experiments. When unilateral lesions included the rostral part of the hypothalamus, CRF-like immunoreactive (CRFI) ipsilateral fibers in the LS decreased in number. Lesions in other brain regions did not cause alterations in the septal CRFI fibers. These findings suggest that the septal CRFI fibers originate in the rostral part of the hypothalamus. Furthermore, combined HRP and immunohistochemical staining on the same sections demonstrated double-labeled cells in two discrete areas within the rostral hypothalamus: one was the perifornical hypothalamic area (PeF) at the level of the paraventricular hypothalamic nucleus, and the other was the most caudal part of the anterior hypothalamic nucleus (AHc). These findings show that a large proportion of the CRFI projections to the LS have their origins in the PeF and AHc.

Ultrastructural evidence for a functional unit between nerve fibers and type II cerebral mast cells in the cerebral vascular wall.

By histofluorescence microscopic examinations of pial arteries from rats and rabbits, we have observed that the routes of adrenergic fibers were apparently organized along successive sites of granular autofluorescent cells present in the adventitia. Subsequent electron microscopic studies showed that these cells were often situated in close apposition (80 to 200 nm) to the adventitial nerve bundles. The granular cells and nerve varicosities were frequently enclosed within the same basement membrane, with a membrane-to-membrane distance as small as 20 nm. However, no clear membrane differentiation was seen. These granular cells were identified histochemically by staining with Sudan Black, Oil Red O, Toluidine Blue, Alcian Blue, together with ultrastructural and pharmacological methods (48/80 compound and carbachol intracarotid infusions). The cells, many of which contained large amounts of lipids, showed morphological ultrastructural and pharmacological similarities to peripheral mast cells. Nerve bundles contained two types of varicosities: some of them degenerated after superior cervical ganglionectomy and were thus of sympathetic origin, whereas the others contained small clear vesicles (probably cholinergic) and/or large dense-cored vesicles (probably peptidergic). As we have shown that cholinomimetics induce exocytosis of these granular cells, the close relationship between these cells and the nerve fibers may indicate a neurogenic control of the cerebrovascular mast cell secretion. As these cells contain potent vasoactive substances, this relationship may be of importance in the genesis of physiological or pathological cerebrovascular events which are, as yet, poorly understood.

Quantification of the intestinal peptide-containing innervation: length density of nerve fibers and total length of nerve supply to the single villus/crypt unit.

The application of a morphometric method to the quantification of peptide-containing nerves in the gut is described. It allows a simple estimation of the nerve fiber supply per unit volume of tissue (length density) and the calculation of the total nerve fiber supply per unit of intestinal mucosa (villus/crypt unit).

Some mammalian retinae lack FMRF-amide-like immunoreactive efferents.

The retinae of a monkey, cat, rat and mouse were examined for FMRF-amide immunoreactivity, a marker for efferent fibers in the retinae of lower vertebrates. No FMRF-amide immunoreactive retinal fibers were found despite their presence in the rat CNS. Therefore, we conclude that efferent fibers, if present in mammalian retinae, are immunologically different from efferent fibers in lower vertebrates.

Co-localization of neuropeptide tyrosine (NPY) and its C-terminal flanking peptide (C-PON).

Neuropeptide tyrosine (NPY) is one of the most abundant and widespread peptides in the mammalian nervous system. Recent isolation and sequencing of the DNA encoding NPY has predicted the existence of a 97 amino acid precursor peptide. Proteolytic processing of this precursor could yield three separate peptide products, an N-terminal signal peptide, neuropeptide tyrosine and a 30 amino acid C-terminal flanking peptide (C-PON). Here, we present evidence that the predicted C-flanking peptide of NPY is widely distributed in both the central and peripheral nervous systems of several mammalian species including man, and has an identical distribution to NPY. It was also demonstrated, using correlative light microscopic immunostaining on serial sections and double electron microscopic immunocytochemistry, that C-PON and NPY immunoreactivities are co-localized in neuronal cell bodies of the brain cortex, sympathetic ganglion cells, norepinephrine-containing granules of the adrenal medulla and in human pheochromocytoma tumor cells.

Distribution and origin of vasoactive intestinal polypeptide-like immunoreactive fibers in the central amygdaloid nucleus of the rat: an immunocytochemical analysis.

We studied the distribution of vasoactive intestinal polypeptide-like immunoreactive (VIPLI) fibers in the central amygdaloid (AC) nucleus of the rat, using indirect immunofluorescence and the origins of such fibers using a combination of retrograde tracing with immunocytochemistry. VIPLI fibers formed a dense fiber plexus in the lateral subdivision of the AC nucleus, but other subdivisions showed little immunoreactivity. Destruction of the supramammillary (SuM) region and the adjacent lateral hypothalamus, both of which contained a group of VIPLI neurons, resulted in the marked reduction of VIPLI fibers in the ipsilateral AC nucleus, indicating that many of the fibers in the AC nucleus originate from these two areas. This assumption was supported by the finding that injection of fast blue dye into the AC nucleus labeled the VIPLI neurons in the SuM region and lateral hypothalamus.

Possible origins of cerebrovascular nerve fibers showing vasoactive intestinal polypeptide-like immunoreactivity: an immunohistochemical study in the dog.

Changes of vasoactive intestinal polypeptide-like immunoreactivity (VIP-LI) in perivascular nerve fibers of the major cerebral arteries were examined immunohistochemically in the dog. The density of cerebrovascular nerve fibers showing VIP-LI (the average number of nerve fibers with VIP-LI in a unit area of the major cerebral arteries) was estimated, by using whole-mount preparations after extirpation of the pterygopalatine, otic or superior cervical ganglion. After pterygopalatine ganglionectomy, the density was markedly decreased in major cerebral arteries of both anterior circulation (the anterior cerebral and middle cerebral arteries) and posterior circulation (the basilar, superior cerebellar, posterior cerebral and posterior communicating arteries). After otic ganglionectomy, the density was moderately reduced in the major arteries of the posterior circulation, but was not decreased in those of the anterior circulation. After superior cervical ganglionectomy, the density was decreased markedly in the major cerebral arteries of the posterior circulation, and moderately in those of the anterior circulation. The results also indicate that the pterygopalatine, otic and superior cervical ganglia supply perivascular nerve fibers showing VIP-LI to the major cerebral arteries bilaterally with an ipsilateral dominance.