Biomedical subjects

I L Gibbins

Publications and source records attributed to I L Gibbins.

At least 19 recordsLinked to original sources

PubMed2004

Differential expression of calcium channels in sympathetic and parasympathetic preganglionic inputs to neurons in paracervical ganglia of guinea-pigs.

Neurons in pelvic ganglia receive nicotinic excitatory post-synaptic potentials (EPSPs) from sacral preganglionic neurons via the pelvic nerve, lumbar preganglionic neurons via the hypogastric nerve or both. We tested the effect of a range of calcium channel antagonists on EPSPs evoked in paracervical ganglia of female guinea-pigs after pelvic or hypogastric nerve stimulation. omega-Conotoxin GVIA (CTX GVIA, 100 nM) or the novel N-type calcium channel antagonist, CTX CVID (100 nM) reduced the amplitude of EPSPs evoked after pelvic nerve stimulation by 50-75% but had no effect on EPSPs evoked by hypogastric nerve stimulation. Combined addition of CTX GVIA and CTX CVID was no more effective than either antagonist alone. EPSPs evoked by stimulating either nerve trunk were not inhibited by the P/Q calcium channel antagonist, omega-agatoxin IVA (100 nM), nor the L-type calcium channel antagonist, nifedipine (30 microM). SNX 482 (300 nM), an antagonist at some R-type calcium channels, inhibited EPSPs after hypogastric nerve stimulation by 20% but had little effect on EPSPs after pelvic nerve stimulation. Amiloride (100 microM) inhibited EPSPs after stimulation of either trunk by 40%, while nickel (100 microM) was ineffective. CTX GVIA or CTX CVID (100 nM) also slowed the rate of action potential repolarization and reduced afterhyperpolarization amplitude in paracervical neurons. Thus, release of transmitter from the terminals of sacral preganglionic neurons is largely dependent on calcium influx through N-type calcium channels, although an unknown calcium channel which is resistant to selective antagonists also contributes to release. Release of transmitter from lumbar preganglionic neurons does not require calcium entry through either conventional N-type calcium channels or the variant CTX CVID-sensitive N-type calcium channel and seems to be mediated largely by a novel calcium channel.

Action Potentials↗

PubMed2001

Neuropeptide Y immunoreactivity in cutaneous sympathetic and sensory neurons during development of the guinea pig.

Different levels of the cutaneous vasculature are innervated selectively by subpopulations of sympathetic neurons distinguished by the presence or absence of immunoreactivity (-IR) for neuropeptide Y (NPY). This study used multiple-labelling immunohistochemistry to examine the appearance of NPY-IR in neurons innervating cutaneous vessels in the ear pinna of embryonic, fetal, and neonatal guinea pigs. NPY-immunoreactive axons were detected in the ear bud at embryonic day 25. However, these axons lacked IR for tyrosine hydroxylase (TH) and often ran in bundles with substance P (SP)-immunoreactive axons close to the epidermis. Many neuronal somata in the cervical dorsal root ganglia (DRG) at late embryonic stages contained NPY-IR with or without SP-IR, but no NPY-IR was detected in DRG or subepidermal axons by late fetal stages. IR for calcitonin gene-related peptide increased in DRG neurons from midfetal to late fetal stages, after the decrease in NPY-IR. Populations of TH-IR neurons with or without NPY-IR were present in the superior cervical ganglion (SCG) from midembryonic stages. TH-immunoreactive axons were not detected in the ear pinna until midfetal stages, when axons with TH-IR and NPY-IR innervated proximal arteries and TH-immunoreactive axons without NPY-IR innervated distal vessels. Vasoactive intestinal peptide-IR was detected transiently in most fetal SCG neurons with TH-IR and NPY-IR but was not detected in cutaneous axons. These results demonstrate that selective expression of NPY by subpopulations of sympathetic neurons occurs prior to innervation of their targets. This suggests that target contact is not required to establish appropriate patterns of expression of peptide neurotransmitters by cutaneous sympathetic neurons.

Aging↗

PubMed2001

Neurochemical differentiation of functionally distinct populations of autonomic neurons.

The coeliac ganglion of guinea pigs displays a unique topographical arrangement of neurochemically and functionally distinct populations of sympathetic neurons. The authors used multiple-labeling immunohistochemistry to investigate the neurochemical differentiation of these neurons during embryonic and fetal development. Sympathoadrenal precursors, located on either side of the abdominal aorta, were intensely immunoreactive for tyrosine hydroxylase (TH-IR), neurofilament, and the human natural killer 1 antibody at midembryonic stages (Carnegie stages 16-19). During late embryonic stages (stages 20-23), a single bilobed ganglion had formed. At this time, neuropeptide Y immunoreactivity (NPY-IR) was widely expressed in sympathetic neurons (with moderate TH-IR) and chromaffin cells (with intense TH-IR). The onset of somatostatin (Som-IR) expression followed that of NPY-IR and was restricted to sympathetic neurons. However, at late embryonic stages, most TH-IR neurons with Som-IR also expressed NPY-IR (a combination of peptides not found in the mature coeliac ganglion). Between late embryonic stages and the end of the early fetal period, there was a significant increase in the proportion of neurons in lateral regions that had both NPY-IR and TH-IR. At the same time, there was an increase in the proportion of neurons in medial regions that had both Som-IR and TH-IR. Neurons expressing both Som-IR and TH-IR were rarely observed in lateral regions of the coeliac ganglion. Thus, a clear topography within the coeliac ganglion is established during late embryonic and early fetal stages of development and reflects that found in the mature animal by the end of the early fetal period.

Adrenal Glands↗

PubMed2001

Expression of immunoreactivity to neurokinin-1 receptor by subsets of cranial parasympathetic neurons: correlation with neuropeptides, nitric oxide synthase, and pathways.

We examined the patterns of coexistence of immunoreactivity to the neurokinin-1 (NK(1)) tachykinin receptor, nitric oxide synthase, and neuropeptides in the sphenopalatine and otic ganglia of guinea pigs using a combination of multiple-labeling immunohistochemistry and pathway tracing in vitro. Most neurons had immunoreactivity to vasoactive intestinal peptide (85-96%) and neuropeptide Y (60%). Subpopulations of vasoactive intestinal peptide-immunoreactive neurons also had immunoreactivity to nitric oxide synthase (37-48%) or enkephalin (25-35%), but these formed mutually exclusive populations. Almost all neurons expressing NK(1) receptor immunoreactivity contained immunoreactivity to enkephalin, vasoactive intestinal peptide, and neuropeptide Y, but not nitric oxide synthase. Using a combination of retrograde axonal tracing and axonal crushing, we found that most neurons with immunoreactivity to nitric oxide synthase projected along the nasopalatine and ethmoidal nerves to the nasal mucosa. In contrast, most neurons with immunoreactivity to enkephalin followed the zygomatic nerve to the facial skin and lacrimal gland. Based on their peptide content, we conclude that the neurons with immunoreactivity to enkephalin and NK(1) receptor projected selectively to the skin. In both the sphenopalatine and the otic ganglia, about half of the neurons with NK(1) receptor immunoreactivity were surrounded by varicose nerve fibers with substance P immunoreactivity. Many of these fibers are likely to have originated in the trigeminal ganglion. Taken together, these observations establish a strong anatomical basis for a range of interactions between trigeminal and cranial parasympathetic pathways that may underlie pathophysiological conditions such as trigeminal neuralgia.

Animals↗

PubMed2001

Differential inhibition by botulinum neurotoxin A of cotransmitters released from autonomic vasodilator neurons.

The role of the soluble NSF attachment protein receptor (SNARE) protein complex in release of multiple cotransmitters from autonomic vasodilator neurons was examined in isolated segments of guinea pig uterine arteries treated with botulinum neurotoxin A (BoNTA; 50 nM). Western blotting of protein extracts from uterine arteries demonstrated partial cleavage of synaptosomal-associated protein of 25 kDa (SNAP-25) to a NH2-terminal fragment of approximately 24 kDa by BoNTA. BoNTA reduced the amplitude (by 70-80%) of isometric contractions of arteries in response to repeated electrical stimulation of sympathetic axons at 1 or 10 Hz. The amplitude of neurogenic relaxations mediated by neuronal nitric oxide (NO) was not affected by BoNTA, whereas the duration of peptide-mediated neurogenic relaxations to stimulation at 10 Hz was reduced (67% reduction in integrated responses). In contrast, presynaptic cholinergic inhibition of neurogenic relaxations was abolished by BoNTA. These results demonstrate that the SNARE complex has differential involvement in release of cotransmitters from the same autonomic neurons: NO release is not dependent on synaptic vesicle exocytosis, acetylcholine release from small vesicles is highly dependent on the SNARE complex, and neuropeptide release from large vesicles involves SNARE proteins that may interact differently with regulatory factors such as calcium.

Acetylcholine↗

PubMed2001

Differential expression of functionally identified and immunohistochemically identified NK(1) receptors on sympathetic neurons.

We have used multiple-labeling immunohistochemistry, intracellular dye-filling, and intracellular microelectrode recordings to characterize the distribution of tachykinin receptors and substance P boutons on subpopulations of neurons within the guinea pig celiac ganglion. Superfusion of substance P (SP, 1 microM for 1 min) depolarized 42% of tonic neurons and inhibited afterhyperpolarizations in 66% of long afterhyperpolarizing (LAH) neurons without significant desensitization. Twenty-one percent of tonic neurons and 24% of LAH neurons responded to the NK(3) agonist senktide but did not respond to SP, indicating SP did not activate NK(3) receptors at this concentration. All effects of SP were abolished by the selective NK(1) receptor antagonist, SR140333, but not by the selective NK(3) receptor antagonist, SR142801, suggesting that exogenous SP activated a receptor with NK(1) pharmacology. No dye-filled LAH neuron and only 50% of tonic neurons responding to SP expressed NK(1) receptor immunoreactivity (NK(1)-IR). All neurons responding to SP had SP immunoreactive fibers within one cell diameter, indicating good spatial matching between SP release sites and target neurons. These results indicate that SP may act via a receptor with NK(1)-like pharmacology that has a C terminus not recognized by antibodies to the intracellular domain of the conventional NK(1) receptor. Inward currents evoked by SP acting on this NK(1)-like receptor or senktide acting through NK(3) receptors had identical current-voltage relationships. In LAH neurons, both agonists suppressed I(sAHP) without reducing I(AHP). Responses evoked by SP and senktide were resistant to PKC inhibitors, suggesting that the transduction mechanisms for the NK(1)-like receptor and the NK(3) receptor may be similar.

Action Potentials↗

PubMed2001

Development of electrophysiological and morphological diversity in autonomic neurons.

The generation of neuronal diversity requires the coordinated development of differential patterns of ion channel expression along with characteristic differences in dendritic geometry, but the relations between these phenotypic features are not well known. We have used a combination of intracellular recordings, morphological analysis of dye-filled neurons, and stereological analysis of immunohistochemically labeled sections to investigate the development of characteristic electrical and morphological properties of functionally distinct populations of sympathetic neurons that project from the celiac ganglion to the splanchnic vasculature or the gastrointestinal tract of guinea pigs. At early fetal stages, neurons were significantly more depolarized at rest compared with neurons at later stages, and they generally fired only a single action potential. By mid fetal stages, rapidly and slowly adapting neurons could be distinguished with a topographic distribution matching that found in adult ganglia. Most rapidly adapting neurons (phasic neurons) at this age had a long afterhyperpolarization (LAH) characteristic of mature vasomotor neurons and were preferentially located in the lateral poles of the ganglion, where most neurons contained neuropeptide Y. Most early and mid fetal neurons showed a weak M current, which was later expressed only by rapidly-adapting and LAH neurons. Two different A currents were present in a subset of early fetal neurons and may indicate neurons destined to develop a slowly adapting phenotype (tonic neurons). The size of neuronal cell bodies increased at a similar rate throughout development regardless of their electrical or neurochemical phenotype or their topographical location. In contrast, the rate of dendritic growth of neurons in medial regions of the ganglion was significantly higher than that of neurons in lateral regions. The apparent cell capacitance was highly correlated with the surface area of the soma but not the dendritic tree of the developing neurons. These results demonstrate that the well-defined functional populations of neurons in the celiac ganglion develop their characteristic electrophysiological and morphological properties during early fetal stages of development. This is after the neuronal populations can be recognized by their neurochemical and topographical characteristics but long before the neurons have finished growing. Our data provide strong circumstantial evidence that the development of the full phenotype of different functional classes of autonomic final motor neurons is a multi-step process likely to involve a regulated sequence of trophic interactions.

Action Potentials↗

PubMed2000

Different levels of immunoreactivity for synaptosomal-associated protein of 25 kDa in vasoconstrictor and vasodilator axons of guinea-pigs.

Immunoreactivity (IR) for synaptosomal-associated protein of 25 kDa (SNAP-25) was examined in axons of autonomic vasoconstrictor and vasodilator neurons innervating the lingual and uterine arteries of guinea-pigs. Polyacrylamide gel electrophoresis and immunoblotting of protein extracts demonstrated a SNAP-25-IR band at 25 kDa in both arteries. Quantitative confocal microscopy demonstrated significantly higher levels of SNAP-25-IR in varicosities with IR for vasoactive intestinal peptide (VIP) than in adjacent axons with IR for tyrosine hydroxylase (TH). Levels of SNAP-25-IR in TH-IR axons, relative to adjacent VIP-IR axons, were significantly higher in the lingual artery than the uterine artery. These differences in IR for SNAP-25, a protein considered essential for calcium-dependent exocytosis of neurotransmitters, raise the possibility that mechanisms of transmitter release may vary between different classes of autonomic neurons.

Animals↗

PubMed2000

Pathway specific expression of neuropeptides and autonomic control of the vasculature.

In this article, we review the immunohistochemical evidence for the pathway-specific expression of co-existing neuropeptides in autonomic vasomotor neurons, and examine the functional significance of these expression patterns for the autonomic regulation of the vasculature. Most final motor neurons in autonomic vasomotor pathways contain neuropeptides in addition to non-peptide co-transmitters such as catecholamines, acetylcholine and nitric oxide. Neuropeptides also occur in preganglionic vasomotor neurons. The precise combinations of neuropeptides expressed by neurons in vasomotor pathways vary with species, vascular bed, and the level within the vascular bed. This applies to both vasoconstrictor and vasodilator pathways. There is a similar degree of variation in the expression of neuropeptide receptors in the vasculature. Consequently, the contributions of different peptides to autonomic vasomotor control are closely matched to the functional requirements of specific vascular beds. This arrangement allows for a high degree of precision in vascular control in normal conditions and has the potential for considerable plasticity under pathophysiological conditions.

Animals↗

PubMed2000

Neuronal morphology and the synaptic organisation of sympathetic ganglia.

In this article, we provide a short review of the structure and synaptic organisation of the final motor neurons in the sympathetic ganglia of mammals. Combinations of pathway tracing, multiple-labelling immunofluorescence and intracellular dye injection have shown that neurons in different functional pathways differ not only in their patterns of neuropeptide expression, but also in the size of their cell bodies and dendritic fields. Thus, vasoconstrictor neurons consistently are smaller than any other major functional class of neurons. Serial section ultrastructural analysis of dye filled neurons, together with electron microscopic and confocal microscopic analysis of immunolabelled synaptic inputs to sympathetic final motor neurons indicate that synapses are rare and randomly distributed over the surface of the neurons. The total number of synapses is simply proportional to the total surface area of the neurons. Many terminal boutons of peptide-containing preganglionic neurons do not make conventional synapses with target neurons. Furthermore, there is a spatial mismatch in the distribution of peptide-containing terminals and neurons expressing receptors for the corresponding peptides. Together, these results suggest that there are likely to be significant differences in the ways that the final sympathetic motor neurons in distinct functional pathways integrate their synaptic inputs. In at least some pathways, heterosynaptic actions of neuropeptides probably contribute to subtle modulation of ganglionic transmission.

Animals↗

PubMed1999

Neurokinin-1 receptor localisation in guinea pig autonomic ganglia.

We have used multiple-labelling immunohistochemistry and confocal microscopy to determine the distribution of immunoreactivity to the tachykinin neurokinin-1 (NK(1)) receptors in guinea pig sympathetic ganglia. Although nerve fibres containing immunoreactivity to substance P were common in all ganglia except the superior cervical ganglia, most neurons expressing NK(1) receptor immunoreactivity were not closely surrounded by pericellular baskets of substance P-immunoreactive boutons. Conversely, many neurons surrounded by baskets of substance P-immunoreactive boutons lacked NK(1) immunoreactivity. In the coeliac and inferior mesenteric ganglia, NK(1) receptor expression was restricted almost entirely to noradrenergic neurons that contained somatostatin immunoreactivity and projected to the enteric plexuses. In the lumbar chain and paracervical ganglia, NK(1) immunoreactivity was expressed by nonnoradrenergic vasodilator neurons containing immunoreactivity to vasoactive intestinal peptide. Taken together, our results show that sympathetic neurons in different functional pathways express NK(1) receptor immunoreactivity. However, the neurons that could respond to endogenously released substance P through NK(1) receptors may be distant from presynaptic release sites. These observations suggest that, in sympathetic ganglia, substance P may modulate ganglionic transmission through heterosynaptic actions on NK(1) receptors.

Animals↗

PubMed1999

Subpopulations of sympathetic neurons project to specific vascular targets in the pinna of the rabbit ear.

We have characterised sympathetic neurons projecting to a range of cutaneous and striated muscle vascular targets in the pinna of the rabbit ear by examining neurotransmitter-related enzymes and peptides in perivascular axons and in somata identified by retrograde axonal tracing. Fast Blue was injected into one of seven sites in each pinna (n = 21 pinnae). The soma cross-sectional area and immunoreactivity (IR) for tyrosine hydroxylase (TH) and neuropeptide Y (NPY) were determined for each of 2,041 retrogradely labelled neurons in the ipsilateral superior cervical ganglion (SCG) or stellate ganglion (StG). Larger neurons in the SCG with TH-IR but not NPY-IR projected predominantly to veins along the medial edge of the pinna. Larger neurons in the StG with TH-IR but not NPY-IR projected predominantly to arteries and veins in the tip and lateral edge of the pinna. Smaller neurons in the SCG with IR to both TH and NPY projected predominantly to arteries in the striated muscles at the base of the ear. The smallest retrogradely labelled neurons in the SCG or StG lacked TH-IR but contained NPY-IR and projected almost exclusively to arterial vessels in the lateral muscle at the base of the ear. Thus, somata of sympathetic neurons projecting to cutaneous versus striated muscle vessels or to different regions of the cutaneous bed could be distinguished by a combination of location, size, and immunohistochemical profile. Consequently, regulation of blood flow within the rabbit ear is likely to involve coordination between neuronal pathways containing neurochemically and morphologically distinct populations of sympathetic neurons.

Animals↗

PubMed1999

Pathway-specific expression of PKC and PKA in sympathetic neurons.

We used multiple-labelling immunofluorescence, intracellular dye injection, electrophysiological recording and confocal microscopy to examine the expression of immunoreactivity to protein kinase C (PKC) and protein kinase A (PKA) in sympathetic ganglia of guinea-pigs. PKCalpha and PKCgamma were widespread in vasoconstrictor and pilomotor neurons. High levels of PKA RIIalpha and RIIbeta were restricted to neurons that lacked significant expression of PKC, including somatostatin-containing neurons projecting to the gut, and non-noradrenergic vasodilator neurons. In coeliac ganglia, most neurons with PKC contained neuropeptide Y and displayed phasic patterns of action potential firing, often with a long after-hyperpolarization. Tonically firing neurons lacked both neuropeptide Y and PKC. These results show remarkably pathway-specific expression of protein kinases in functionally identified populations of sympathetic neurons.

Action Potentials↗

PubMed1999

Electrophysiological and morphological diversity of mouse sympathetic neurons.

We have used multiple-labeling immunohistochemistry, intracellular dye-filling, and intracellular microelectrode recordings to characterize the morphological and electrical properties of sympathetic neurons in the superior cervical, thoracic, and celiac ganglia of mice. Neurochemical and morphological characteristics of neurons varied between ganglia. Thoracic sympathetic ganglia contained three main populations of neurons based on differential patterns of expression of immunoreactivity to tyrosine hydroxylase, neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP). In the celiac ganglion, nearly all neurons contained immunoreactivity to both tyrosine hydroxylase and NPY. Both the overall size of the dendritic tree and the number of primary dendrites were greater in neurons from the thoracic and celiac ganglia compared with those from the superior cervical ganglion. The electrophysiological properties of sympathetic neurons depended more on their ganglion of origin rather than their probable targets. All neurons in the superior cervical ganglion had phasic firing properties and large afterhyperpolarizations (AHPs). In addition, 34% of these neurons displayed an afterdepolarization preceding the AHP. Superior cervical ganglion neurons had prominent I(M), I(A), and I(H) currents and a linear current-voltage relationship between -60 and -110 mV. Neurons from the thoracic ganglia had significantly smaller action potentials, AHPs, and apparent cell capacitance compared with superior cervical ganglion neurons, and only 18% showed an afterdepolarization. All neurons in superior cervical ganglia and most neurons in celiac ganglia received at least one strong preganglionic input. Nearly one-half the neurons in the celiac ganglion had tonic firing properties, and another 15% had firing properties intermediate between those of tonic and phasic neurons. Most celiac neurons showed significant inward rectification below -90 mV. They also expressed I(A), but with slower inactivation kinetics than that of superior cervical or thoracic neurons. Both phasic and tonic celiac ganglion neurons received synaptic inputs via the celiac nerves in addition to strong inputs via the splanchnic nerves. Multivariate statistical analysis revealed that the properties of the action potential, the AHP, and the apparent cell capacitance together were sufficient to correctly classify 80% of neurons according to their ganglion of origin. These results indicate that there is considerable heterogeneity in the morphological, neurochemical, and electrical properties of sympathetic neurons in mice. Although the morphological and neurochemical characteristics of the neurons are likely to be related to their peripheral projections, the expression of particular electrophysiological traits seems to be more closely related to the ganglia within which the neurons occur.

Action Potentials↗

PubMed1998

Synaptic organisation of lumbar sympathetic ganglia of guinea pigs: serial section ultrastructural analysis of dye-filled sympathetic final motor neurons.

The authors serially sectioned seven dye-filled neuronal somata and more than 1.6 mm of their dendrites from the lumbar sympathetic ganglia of guinea pigs and examined them ultrastructurally to determine the distribution of preganglionic synaptic inputs to their dendrites and cell bodies. Most of the surface of the neurons was covered with Schwann cells. Apposing boutons were rare, with an average density of one axosomatic bouton per 125 microm2 of somatic membrane and one axodendritic bouton per 25 microm of dendrite. Many dendritic segments that were more than 50 microm long completely lacked any apposing boutons. Although the average density of apposing boutons was low, local densities could be high, so that clusters of up to four adjacent boutons occurred on cell bodies and dendrites alike. The spatial arrangement of the apposing boutons for each of the cells examined here was not significantly different from a random distribution. Consequently, the number of apposing boutons observed for any neuron was simply proportional to the amount of neuronal surface sampled in the serial section run. About 50% of boutons directly apposing the neurons lacked any detectable presynaptic specialisations. When they were present, the presynaptic densities had a mean length of about 220 nm, with no difference between boutons that made axosomatic or axodendritic appositions. By applying these data to complete reconstructions of the dendritic trees of dye-filled sympathetic neurons at the light microscopic level, the authors estimated that few neurons in the lumbar sympathetic chain of guinea pigs would receive more than 200 synapses or apposing boutons and that many of them would receive less than 100 synapses. Up to 50% of these boutons would be predicted to make axosomatic contacts. These new observations provide a strong morphological framework for a better understanding of how sympathetic final motor neurons process their preganglionic synaptic inputs.

Animals↗

PubMed1998

Synaptic organisation of neuropeptide-containing preganglionic boutons in lumbar sympathetic ganglia of guinea pigs.

Within the lumbar sympathetic ganglia of guinea pigs, the endings of different populations of neuropeptide-containing preganglionic neurons form well-defined pericellular baskets of boutons around target neurons in specific functional pathways. We have used multiple-labelling immunofluorescence, confocal microscopy, and ultrastructural immunocytochemistry to investigate synaptic organisation within pericellular baskets labelled for immunoreactivity to calcitonin gene-related peptide (CGRP), substance P (SP), or the pro-enkephalin-derived peptide, met-enkephalin-arg-gly-leu (MERGL) in relation to their target neurons. Different functional populations of neurons, identified by their neurochemical profile, showed a significant degree of spatial clustering and predicted well the distribution of specific classes of pericellular baskets. Most of the boutons in a basket were completely surrounded by Schwann cell processes and did not form synapses. The synapses that were present were made mostly onto dendrites enclosed by the Schwann cell sheath surrounding the neuron within the basket. These dendrites probably originated from neurochemically similar neighbouring neurons. Nevertheless, some of the boutons in the baskets did form synapses with the cell body or proximal dendrites of the neuron they surrounded. Occasionally, cell bodies received a relatively high number of synapses and close appositions from boutons in a pericellular basket. Synaptic convergence of two immunohistochemically distinct types of preganglionic inputs was found in baskets of SP-immunoreactive or MERGL-immunoreactive, but not CGRP-immunoreactive, boutons. Taken together, our results show that the appearance of pericellular baskets is primarily due to the packing of the target neurons. The grouping of functionally similar classes of neurons with their pathway-specific projections of peptide-containing preganglionic neurons suggests that peptides could exert their effects in relatively well-defined zones within the ganglia.

Animals↗

PubMed1998

Neurochemical distinction between skeletal muscle vasodilator neurons and pelvic vasodilator neurons in guinea-pigs.

This study sets out to compare the combinations of potential vasodilator transmitters expressed by sympathetic and pelvic vasodilator neurons of guinea-pigs. Triple-labelling fluorescence immunohistochemistry was used to examine immunoreactivity (IR) to vasoactive intestinal peptide (VIP), nitric oxide synthase (NOS) and calcitonin gene-related peptide (CGRP) in lumbar sympathetic ganglia, and in perivascular axons supplying hindlimb skeletal muscles or pelvic viscera. Only 0.2% of VIP-IR nerve cell bodies in lumbar sympathetic ganglia (n = 4632 VIP-IR nerve cell profiles) contained NOS-IR, and one VIP-IR neuron contained CGRP-IR. The VIP-IR perivascular axons along the common and external iliac arteries, femoral artery and arteries to hindlimb muscles lacked NOS-IR and CGRP-IR. In contrast, all VIP-IR perivascular axons projecting from pelvic ganglia to the main uterine artery, and half of the VIP-IR axons along the internal iliac artery, contained NOS-IR and CGRP-IR. Thus, the neurochemical content of sympathetic vasodilator neurons to skeletal muscle arteries was clearly distinguishable from that of pelvic vasodilator neurons to the uterine vasculature. Furthermore, the autonomic dilation in each vascular bed is likely to be qualitatively different, and matched to the functional requirements of each target organ.

Animals↗

PubMed1998

Peripheral fields of sympathetic vasoconstrictor neurons in guinea pigs.

We have combined retrograde axonal tracing using Fast Blue and Dil, with immunohistochemistry, to estimate the maximum size of peripheral fields of identified sympathetic vasoconstrictor neurons projecting to guinea-pig ear tips. Many neurons in the superior cervical ganglia were labelled with both Fast Blue and Dil after dye injections up to 7 mm apart. Few neurons were labelled when dye injections were 8-10 mm apart. Neurons labelled with both Dil and Fast Blue after dye injections 5-7 mm apart had, on average, larger somata (436 +/- 84 microm2, mean +/- SEM, n = 47) than neurons labelled with Dil only (388 +/- 11 microm2, n = 147). Typically, 50-100 neurons innervated a region of vasculature 1 mm in diameter. We conclude that sympathetic vasoconstrictor neurons branch widely before converging on to their target blood vessels. Progressive recruitment of vasoconstrictor neurons with increasing field size would provide an efficient mechanism for graded neural control of the circulation.

Amidines↗