vasoactive-intestinal-peptide and true-blue

The origin of perivascular nerve fibres storing nitric oxide synthase (NOS) and co-localisation with perivascular neuropeptides were examined in the rat middle cerebral artery (MCA) by retrograde tracing with True Blue (TB) in combination with immunocytochemistry. Application of TB to the proximal part of the middle cerebral artery labelled nerve cell bodies ipsilaterally in the trigeminal, sphenopalatine, otic, and superior cervical ganglia. A few labelled cell bodies were seen contralaterally, suggesting bilateral innervation. In the parasympathetic sphenopalatine and otic ganglia, numerous TB-labelled cell bodies contained neuronal NOS (C- and N-terminal), vasoactive intestinal peptide (VIP), and pituitary adenylate cyclase activating peptide (PACAP). In the trigeminal ganglion, almost all TB-labelled cell bodies contained calcitonin gene-related peptide (CGRP) but only a few cells contained NOS. In the superior cervical ganglion, the majority of the TB-labelled nerve cells contained neuropeptide Y (NPY) but none of them contained NOS. Removal of the ipsilateral sphenopalatine ganglion caused a slight reduction in the number of perivascular VIP-, PACAP-, and NOS-containing fibres after 3 days in the MCA while there was no difference at 2 and 4 weeks after the denervation as compared to control. This indicates that the parasympathetic VIP-, PACAP-, and NOS-immunoreactive nerve fibres in the rat MCA originate from several sources.

Topics: Animals; Benzofurans; Calcitonin Gene-Related Peptide; Cerebral Arteries; Denervation; Fluorescent Antibody Technique; Fluorescent Dyes; Ganglia, Parasympathetic; Immunohistochemistry; Male; Middle Cerebral Artery; Nerve Fibers; Neuropeptides; Nitric Oxide Synthase; Pituitary Adenylate Cyclase-Activating Polypeptide; Rats; Rats, Sprague-Dawley; Superior Cervical Ganglion; Trigeminal Ganglion; Vasoactive Intestinal Peptide

The distribution and origin of neuropeptide Y-, vasoactive intestinal peptide- and calcitonin gene-related peptide-containing nerve fibers and adrenergic (dopamine-beta-hydroxylase-containing) fibers in the rat larynx were studied by retrograde tracing and selective denervations in combination with immunocytochemistry. An injection of the retrograde tracer True Blue to the right vocal cord resulted in the appearance of labelled nerve cell bodies in the ipsi- and contralateral superior cervical and stellate ganglia, the thyroid ganglia, the jugular-nodose ganglionic complexes, in the ipsilateral trigeminal and dorsal root ganglia at levels C2 and C3 and in local tracheal ganglia. Judging from the number of labelled nerve cell bodies, the jugular-nodose ganglionic complexes, dorsal root ganglia and superior cervical ganglia provide the greater part of the vocal cord innervation. Most of the True Blue-labelled nerve cell bodies in the superior cervical and stellate ganglia contained neuropeptide Y. In the thyroid ganglia the majority of labelled nerve cell bodies contained vasoactive intestinal peptide. In the jugular-nodose ganglionic complex and the dorsal root ganglia the majority of the labelled nerve cell bodies stored calcitonin gene-related peptide. Retrograde tracing and denervation studies revealed that all noradrenaline- and the majority of neuropeptide Y-containing nerve fibers emanate from the superior cervical and stellate ganglia. A minor population of neuropeptide Y-containing nerve fibers originate in local tracheal ganglia. The vasoactive intestinal peptide-containing nerve fibers originate in the thyroid ganglion and local tracheal ganglia, whereas calcitonin gene-related peptide-containing nerve fibres emanate from the dorsal root ganglia (C2-C3), the trigeminal ganglia and the jugular-nodose ganglia.

Topics: Animals; Benzofurans; Calcitonin Gene-Related Peptide; Dopamine beta-Hydroxylase; Fluorescent Dyes; Immunohistochemistry; Nerve Fibers; Neurons; Neuropeptide Y; Rats; Sympathectomy; Sympathetic Nervous System; Synaptic Transmission; Vasoactive Intestinal Peptide; Vocal Cords

The origin of nerve fibers to the rat middle cerebral artery was studied by retrograde tracing with the fluorescent tracer True Blue (TB) in combination with immunocytochemistry to known perivascular peptides. Application of TB to the middle cerebral artery labeled nerve cell bodies in the ipsilateral superior cervical ganglion, the otic ganglion, the sphenopalatine ganglion, the trigeminal ganglion, and the cervical dorsal root ganglion at level C2. A few labeled nerve cell bodies were seen in contralateral ganglia. Judging from the number and intensity of the labeling, the superior cervical ganglion and the trigeminal ganglion and dorsal root ganglion at level C2 contributed most to the innervation. A moderate number of nerve cell bodies were labeled in the sphenopalatine and otic ganglia. The TB-labeled nerve cell bodies were further examined for the presence of neuropeptides. For that purpose antibodies raised against neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP) and calcitonin gene-related peptide (CGRP) were used. A considerable portion of the TB-labeled nerve cell bodies in the superior cervical ganglion contained NPY. About half of the labeled nerve cell bodies in the sphenopalatine and otic ganglia contained VIP. In the trigeminal ganglion and in the dorsal root ganglion at level C2, one-third of the TB-labeled nerve cell bodies were CGRP-immunoreactive, while only few nerve cell bodies contained SP. The study provides direct evidence for the origin of cerebrovascular peptidergic nerve fibers and demonstrates that not only ipsilateral but also contralateral ganglia contribute to the innervation of the cerebral circulation.

Topics: Animals; Benzofurans; Calcitonin Gene-Related Peptide; Cerebral Arteries; Immunohistochemistry; Nerve Fibers; Neuropeptide Y; Neuropeptides; Rats; Substance P; Vasoactive Intestinal Peptide

In immunohistochemical studies on rat two types of nerve fibres, both showing substance P and calcitonin gene-related peptide-like immunoreactivity, have been localized in the sphenopalatine ganglion, the principal cells of which contain both vasoactive intestinal polypeptide and choline acetyltransferase. One fine-calibre fibre type forms basket-like arrangements around approximately 3-5% of the principal neurons, whereas another, more coarse type traverses the ganglion without making contacts with the ganglion cells. By transection of nerves connecting with the ganglion, in combination with retrograde tracing experiments, it was concluded that the fine-calibre fibres exclusively come from the trigeminal ganglion, whereas the second type in addition, and mainly, originate in the internal carotid ganglion which is situated along the greater superficial petrosal nerve and the pterygoid nerve at their junction with the internal carotid nerve. The brain vasculature was shown to be one target structure for the innervated principal cells in the sphenopalatine ganglion. The arrangement provides the functional possibility for a modulatory interaction between the autonomic and sensory systems, thus resembling an axon reflex mechanism in the peripheral nervous system.

Topics: Animals; Benzofurans; Calcitonin Gene-Related Peptide; Choline O-Acetyltransferase; Ganglia, Parasympathetic; Immunohistochemistry; Male; Neural Pathways; Neuropeptides; Rats; Rats, Inbred Strains; Substance P; Trigeminal Nerve; Vasoactive Intestinal Peptide

Retrograde neuronal tracing with the fluorescent dye True Blue and immunocytochemistry were utilized to examine postganglionic sympathetic neurons in para- and prevertebral ganglia projecting to the rat ovary. Perikarya in both ganglia were labeled with True Blue after application of the tracer to either the superior ovarian or ovarian plexus nerve. After application of True Blue to the superior ovarian nerve, 17% of the labeled cells in paravertebral ganglia were immunoreactive for vasoactive intestinal polypeptide. In contrast, after application of True Blue to the ovarian plexus nerve, approximately 1% of the labeled cells in paravertebral ganglia were immunoreactive for the same polypeptide. Some vasoactive intestinal polypeptide-immunoreactive perikarya in paravertebral ganglia were not labeled with True Blue. In some cases, substance P- and vasoactive intestinal polypeptide-immunoreactive fibers were closely apposed to True Blue-labeled perikarya in para- and prevertebral ganglia. Paravertebral vasoactive intestinal polypeptide-immunoreactive perikarya projecting to the ovary presumably participate directly in the control of various ovarian functions. Substance P- and vasoactive intestinal polypeptide-immunoreactive fibers closely apposed to perikarya projecting to the ovary may participate indirectly in the control of various ovarian functions by affecting the activity of ovarian postganglionic neurons.

Topics: Animals; Benzofurans; Female; Fluorescent Antibody Technique; Ganglia, Sympathetic; Immunohistochemistry; Nerve Fibers; Neurons; Ovary; Rats; Rats, Inbred Strains; Substance P; Vasoactive Intestinal Peptide

Retrograde tracing of the fluorescent marker, True Blue, has been used together with immunohistochemistry employing antibodies to substance P, calcitonin gene-related peptide, somatostatin, vasoactive intestinal polypeptide and morphine-modulating peptide to study the afferent innervation of the stomach in rat, mouse and guinea-pig. Up to 85% of spinal afferents to the stomach in all three species contained immunoreactive calcitonin gene-related peptide, and up to 50% contained substance P. In all three species less than 10% of vagal afferents to the stomach reacted with antibodies to calcitonin gene-related peptide, or substance P. Cacitonin gene-related peptide-immunoreactive fibres were found in the myenteric plexus, circular muscle and around submucosal blood vessels in the stomach. In the rat, removal of the coeliac ganglion, splanchnic nerve section, or capsaicin treatment virtually abolished calcitonin gene-related peptide immunoreactivity in the stomach. Capsaicin and splanchnic section also abolished the staining of immunoreactive calcitonin gene-related peptide fibres in the coeliac ganglion. The same treatments abolished substance P staining of fibres around submucosal blood vessels, but in the myenteric plexus and circular smooth muscle there were still abundant immunoreactive fibres, presumably arising from intrinsic cell bodies. No somatostatin-containing visceral afferents could be found, although somatostatin was localized to cell bodies in rat dorsal root ganglia. Immunoreactive vasoactive intestinal polypeptide-containing dorsal root ganglia neurons were not found; although antibodies to morphine-modulatory peptide revealed immunoreactive nerve cell bodies, we were unable to exclude the possibility that this result is attributable to cross reactivity with calcitonin gene-related peptide. These results provide direct evidence that calcitonin gene-related peptide is a marker for a major subset of visceral primary afferent neurons and suggest that this population of spinal afferents makes a major contribution to the total gastric content of calcitonin gene-related peptide.

Topics: Afferent Pathways; Animals; Benzofurans; Calcitonin Gene-Related Peptide; Guinea Pigs; Immunohistochemistry; Male; Mice; Neuropeptides; Oligopeptides; Rats; Rats, Inbred Strains; Somatostatin; Stomach; Substance P; Vasoactive Intestinal Peptide

One month after sciatic nerve section, only dorsal root ganglion cells which take up True blue applied to the cut end of the nerve show vasoactive intestinal polypeptide immunoreactivity. This indicates that VIP expression is only in cells with damaged axons. Motor axons with True blue-positive perikarya in the ventral horn originate from the fourth to sixth lumbar segments whereas unmyelinated and small myelinated sensory nerves terminate in the third to fifth lumbar segments of the spinal cord.

Topics: Animals; Axons; Benzofurans; Biological Transport; Denervation; Histocytochemistry; Immunochemistry; Male; Microscopy, Fluorescence; Nerve Tissue Proteins; Rats; Sciatic Nerve; Vasoactive Intestinal Peptide