scholarly journals Cholinergic and VIPergic Innervation in Cerebral Arteries: A Sequential Double-Labeling Immunohistochemical Study

1990 ◽  
Vol 10 (1) ◽  
pp. 32-37 ◽  
Author(s):  
Frederick Jia-Pei Miao ◽  
Tony Jer-Fu Lee
Keyword(s):  
Vasoactive Intestinal Polypeptide ◽  
Arterial Wall ◽  
Immunohistochemical Study ◽  
Picric Acid ◽  
Cerebral Arteries ◽  
Nerve Fibers ◽  
Morphological Data ◽  
Immunohistochemical Method ◽  
Double Labeling ◽  
Functional Interactions

The possible co-localization of choline acetyltransferase (ChAT) and vasoactive intestinal polypeptide (VIP) in the nerve fibers of cat cerebral arteries was examined by a sequential double-labeling immunohistochemical method. Diaminobenzidine and tetramethylbenzidine were used as chromogens to distinguish ChAT (protein) and VIP (peptide) immunoreactivities. Since available fixatives often did not provide simultaneous preservation of optimal protein and peptide immunoreactivities, a new fixative, buffered periodate-paraformal-dehyde-picric acid-formaldehyde-lysine (PPPFL), was formulated and tested. PPPFL fixative is more reliable for simultaneously preserving ChAT and VIP immunoreactivities than were periodate-lysine-paraformaldehyde (PLP) fixative, Zamboni's fixative, or 2% paraformaldehyde solution alone. Using PPPFL as fixative, both ChAT immunoreactive (ChAT-I) and VIP-immunoreactive (VIP-I) fibers in cerebral arteries appeared as bundle and fine fibers. Most ChAT-I and VIP-I fibers were separate. Portions of ChAT-I and VIP-I fibers often ran closely in parallel or across each other. Overlaying of VIP-I on ChAT-I fibers and relay connections between them were also observed. These morphological data suggest the potential functional interactions between cholinergic and VIPergic innervations. In <5% of the fibers examined did ChAT and VIP immunoreactivities appear to be co-localized. These data therefore do not support the hypothesis that acetylcholine and VIP are co-localized in most fibers innervating the cerebral arterial wall.

scholarly journals Distribution of serotonin-like immunoreactive nerves in major cerebral arteries of rabbit: an immunohistochemical study.

1989 ◽  
Vol 37 (9) ◽  
pp. 1383-1386 ◽  
Author(s):  
U Dhall ◽  
G Burnstock
Keyword(s):  
Immunohistochemical Study ◽  
Anterior Part ◽  
Cerebral Arteries ◽  
Posterior Part ◽  
Nerve Fibers ◽  
Circle Of Willis ◽  
Immunofluorescence Technique ◽  
Whole Mount ◽  
Peptidergic Innervation ◽  
Indirect Immunofluorescence Technique

We studied the distribution pattern of serotonin-like immunoreactive nerve fibers in the major cerebral vessels of rabbit by an indirect immunofluorescence technique using whole-mount stretch preparations. The density of serotonin-like immunoreactive nerve fibers was greater in vessels of the posterior part of the circle of Willis compared with that in the anterior part. This is in contrast to most of the observations reported previously regarding adrenergic, cholinergic, and peptidergic innervation of the circle of Willis.

scholarly journals Evidence for the Presence of Cholinergic Nerves in Cerebral Arteries: An Immunohistochemical Demonstration of Choline Acetyltransferase

1985 ◽  
Vol 5 (2) ◽  
pp. 327-334 ◽  
Author(s):  
Akira Saito ◽  
Jang-Yen Wu ◽  
Tony J.-F. Lee
Keyword(s):  
Vasoactive Intestinal Polypeptide ◽  
Choline Acetyltransferase ◽  
Cerebral Arteries ◽  
Circle Of Willis ◽  
Immunohistochemical Method ◽  
Cholinergic Nerves ◽  
Vertebral Arteries ◽  
Middle Cerebral Arteries ◽  
Adventitial Layer ◽  
Basilar Arteries

The presence of cholinergic nerves in cerebral arteries of several species was investigated by an immunohistochemical method using antibodies against choline acetyltransferase (ChAT). In cats, pigs, rats, and dogs, ChAT immunoreactivities were found to be associated with large bundles and single fibers in the circle of Willis and anterior cerebral, middle cerebral, and basilar arteries. In the rabbit, the ChAT-immunoreactive (ChAT-I) nerves were also observed in the circle of Willis and anterior and middle cerebral arteries, but only few or none were found in the basilar and vertebral arteries. The ChAT-I nerves were found only in the adventitial layer of vessels examined. Superior cervical ganglionectomy did not appreciably affect the distribution of ChAT-I nerves. These results indicate the presence of cholinergic nerves in cerebral arteries. The distribution pattern of ChAT-I nerves was different from that of vasoactive intestinal polypeptide (VIP)-like-immunoreactive nerves and acetylcholinesterase-positive nerves. The possible coexistence of ChAT and VIP-like substance in the same neuron is discussed.

Demonstration of Neuropeptide Containing Nerves and Vasomotor Responses to Perivascular Peptides in Human Cerebral Arteries

Cephalalgia ◽  
1994 ◽  
Vol 14 (2) ◽  
pp. 88-96 ◽  
Author(s):  
L Edvinsson ◽  
I Jansen ◽  
M Cunha e Sa ◽  
S Gulbenkian
Keyword(s):  
Tyrosine Hydroxylase ◽  
Substance P ◽  
Vasoactive Intestinal Polypeptide ◽  
Cerebral Arteries ◽  
Nerve Fibers ◽  
Neurokinin A ◽  
Vasomotor Responses ◽  
Calcitonin Gene ◽  
Prostaglandin F ◽  
Migraine Pathophysiology

A rich supply of nerve fibers containing neuropeptide Y-like (NPY-LI) and tyrosine hydroxylase-like immunoreactivity was seen in human cerebral arteries, arterioles and veins. Only a sparse supply of vasoactive intestinal polypeptide (VIP-LI), substance P (SP-LI), and calcitonin gene-related peptide (CGRP-LI) was demonstrated in the walls of human cerebral vessels. In isolated ring segments of human cerebral arteries, NPY and noradrenaline caused vasoconstriction but did not potentiate each other. VIP, peptide histidine methionine, SP, neurokinin A, and CGRP relaxed arteries precontracted by prostaglandin F2a. The degree of innervation and the vasomotor responses are discussed in relation to migraine pathophysiology.

scholarly journals Immunohistochemical Localization of the VIP1 Receptor (VPAC1R) in Rat Cerebral Blood Vessels: Relation to PACAP and VIP Containing Nerves

2000 ◽  
Vol 20 (8) ◽  
pp. 1205-1214 ◽  
Author(s):  
Jan Fahrenkrug ◽  
Jens Hannibal ◽  
Jeppe Tams ◽  
Birgitte Georg
Keyword(s):  
Vasoactive Intestinal Polypeptide ◽  
Cerebral Arteries ◽  
Immunohistochemical Localization ◽  
Nerve Fibers ◽  
Receptor Protein ◽  
Receptor Subtypes ◽  
Intimate Contact ◽  
Fluorescence Confocal Microscopy ◽  
The Brain ◽  
Intestinal Polypeptide

The two structurally related peptides, vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP), are present in cerebral vascular nerve fibers. Biologic actions of VIP are exerted through two receptors, VPAC1 and VPAC2, having similar binding affinity for both VIP and PACAP. In the current study, the authors have developed a specific antibody against the rVPAC1 receptor to examine the localization of rVPAC1 immunoreactivity in cerebral arteries and arterioles of the rat by immunohistochemistry using fluorescence confocal microscopy. Specificity of the antiserum was ensured by immunoblotting and immunocytochemistry of cells transfected with cDNA encoding the different PACAP-VIP receptor subtypes. The rVPAC1 receptor immunoreactivity was localized to the plasmalemma of circularly orientated smooth muscle cells on superficial cerebral arteries and arterioles taken from the basal surface of the brain. By double immunostaining VIP immunoreactive nerve fibers and, to a lesser extent, those containing PACAP were shown to have intimate contact with the receptor protein. Vasoactive intestinal polypeptide and PACAP containing cerebrovascular nerve fibers were found in separate nerve populations with different distribution pattern and density. In brain sections processes of cortical VIP-, but not PACAP-, containing neurons seemed to innervate the rVPAC1 receptor of pial arterioles on the brain surface. The current findings provide the neuroanatomical substrate for a role of VIP and maybe PACAP in the regulation of cerebral blood flow.

scholarly journals A reliable method combining horseradish peroxidase histochemistry with immuno-beta-galactosidase staining.

1988 ◽  
Vol 36 (9) ◽  
pp. 1091-1096 ◽  
Author(s):  
M Sakanaka ◽  
S Magari ◽  
T Shibasaki ◽  
K Shinoda ◽  
J Kohno
Keyword(s):  
Cerebral Cortex ◽  
Horseradish Peroxidase ◽  
Pyramidal Cells ◽  
Nerve Fibers ◽  
Immunohistochemical Method ◽  
Double Labeling ◽  
Enzyme Systems ◽  
Pap Method ◽  
Hrp Histochemistry ◽  
Beta Galactosidase

A sensitive combination of horseradish peroxidase (HRP) tracing and immunohistochemistry was used by Rye et al. [J Histochem Cytochem (1984) 32:1145] in a search for the origins of neurotransmitter- and neuromodulator-containing nerve fibers in brain. In this combination, peroxidase as a marker in immunohistochemistry was thought to yield a homogeneous brown immunoreaction product of diaminobenzidine, different from the black granular reaction product of retrogradely transported HRP, which is visualized by the tetramethylbenzidine (TMB) reaction and subsequent stabilization. A neuron that exhibits both kinds of reaction products in its cytoplasm in sections subjected to combination staining is referred to as a double-labeled cell. With a combined HRP and corticotropin-releasing factor (CRF) immunoperoxidase-antiperoxidase (PAP) method, the first set of experiments showed "false" double-labeled cells in the pyramidal cell layer of rat cerebral cortex, but only rarely in the subcortical areas, possibly because of the use of one enzyme system in two different histochemical procedures. This limitation of the double-staining technique prompted us to demonstrate an alternate combination of HRP tracing and immunohistochemistry in the second set of experiments by employing two previously described independent enzyme systems: HRP as a retrograde tracer and beta-galactosidase as a marker for immunohistochemical demonstration of CRF. A homogeneous blue reaction product indicated immuno-beta-galactosidase staining, and a granular black or brown reaction product labeled retrogradely transported HRP in double-labeled cells in subcortical regions. Neither double labeling nor "false" double labeling was seen in pyramidal cells of cerebral cortex. These findings suggest that application of two independent enzyme systems in a combined HRP and immunohistochemical method may be useful for investigating in origins of peptidergic fibers in brain when the combination of HRP histochemistry and the PAP method appears to be inappropriate.

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