[Nerve cells can have more than one signal substance: coexistence of classical transmitters and peptides].
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Biomedical subjects
Publications and source records attributed to M Schultzberg.
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Pieces of embryonic mesencephalic tissue rich in dopamine and cholecystokinin immunoreactive neurones were grafted to the dorsal surface of the caudate-putamen of adult host rats subjected to unilateral dopamine depleting lesions. After 3 months, neuronal survival in the graft and fibre outgrowth into the host brain were studied by tyrosine hydroxylase and cholecystokinin immunohistochemistry, both in serial sections and by elution and restaining of the same sections. Both dopamine- and cholecystokinin-containing neurones as well as neurons containing both compounds survived the transplantation process. The ratio of neurones in which dopamine and cholecystokinin-like immunoreactivity occurred independently and in coexistence was similar in the grafts to that seen in the intact ventral mesencephalon. This suggests that the grafted cells maintain and express at least some of their normal chemical characteristics in the ectopic cortical location. Only those fibres which contained tyrosine hydroxylase but apparently lacked the cholecystokinin-like peptide showed extensive reinnervation of the host neostriatum. The cholecystokinin-positive fibres were found in a narrow zone immediately adjoining the graft. These results indicate that the dopaminergic reinnervation of the denervated neostriatum is preferentially carried out by the population of grafted mesencephalic dopamine neurones apparently lacking the cholecystokinin-like peptide. This suggests the presence of growth regulating mechanisms in the denervated neostriatum which selectively favour the ingrowth of fibres from the appropriate dopaminergic neuronal subset. The transplantation technique may therefore provide a powerful tool for the study of neurone-target interactions in the establishment of neuronal connections, and of the possible role of peptidergic coexistence in the development and organization of monoaminergic pathways and their innervation patterns.
The distribution of dynorphin-like immunoreactivity in the autonomic nervous system of the rat and guinea-pig was investigated using an antiserum raised against dynorphin-(1-17). Dynorphin-like immunoreactivity was observed in fiber networks in the prevertebral sympathetic ganglia, in fibers in the enteric plexa , circular muscle layer and a few in the lamina muscularis of the mucosa of the gastrointestinal tract. After colchicine treatment dynorphin-immunoreactive cell bodies were observed both in the myenteric and submucous ganglia of the gut. The paravertebral ganglia contained occasional dynorphin-positive fibers and a few immunoreactive small intensely fluorescent cells. The distribution of the dynorphin-like immunoreactivity was compared to the distribution of enkephalin-like immunoreactivity, and it was found that in some areas there were similarities in the distribution patterns, although in other areas there were clear differences, indicating a non-identity of these two systems.
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Bombesin-like immunoreactivity occurs in a dense network of nerve fibres in the rat coeliac-superior mesenteric ganglion. No bombesin immunoreactive cell bodies have been observed in this ganglion. An origin outside the ganglion was therefore assumed for the bombesin immunoreactive fibres. Ligation of the mesenteric nerve, between the ganglion and the gut resulted in an accumulation of bombesin immunoreactive material on the distal side, i.e. towards the gut. Transection of the nerve caused a depletion of bombesin immunoreactive fibres in the ganglion. These results suggest that bombesin immunoreactive fibres in the coeliac-superior mesenteric ganglion of the rat originate in neurons in the gastrointestinal wall. Application of colchicine to the stomach wall caused an accumulation of bombesin immunoreactive material in myenteric neurons.
The distribution of substance P immunoreactive nerve fibers in guinea pig sympathetic ganglia was studied in normal and capsaicin-treated animals. A marked depletion of substance P-like immunoreactivity was observed in both pre- and paravertebral ganglia after capsaicin treatment. In the inferior mesenteric and pelvic ganglia, however, intensely fluorescent fibers surrounding some of the principal ganglion cells remain after capsaicin treatment. Ligation experiments on the inferior mesenteric ganglion indicate that the capsaicin-insensitive substance P immunoreactive fibers in this ganglion are of lumbar splanchnic origin. Whether or not they represent capsaicin-resistant primary afferent neurons is not known. The enkephalin, vasoactive intestinal polypeptide, cholecystokinin and bombesin immunoreactive fiber networks in the inferior mesenteric ganglion were also studied, and no differences were observed between normal and capsaicin-treated animals.
After different denervation procedures the guinea-pig inferior mesenteric ganglion was analysed by immunohistochemistry using antisera to substance P, enkephalin, vasoactive intestinal polypeptide, cholecystokinin and bombesin. The results demonstrate that each of the nerve trunks connected to the ganglion carries specific peptidergic pathways. Thus, the lumbar nerves contain substance P-immunoreactive primary afferent neurons, which to a large extent traverse the ganglion, and enkephalin-immunoreactive preganglionic neurons; the colonic nerves carry vasoactive intestinal polypeptide-, cholecystokinin- and bombesin-immunoreactive fibers from the distal colon to the ganglion; the hypogastric nerves contain vasoactive intestinal polypeptide-positive fibers from the pelvic plexus; and the intermesenteric nerve contains vasoactive intestinal polypeptide, cholecystokinin, substance P and enkephalin from divergent sources. By studying accumulations of peptides in ligated lumbar splanchnic, intermesenteric, hypogastric and colonic nerves the existence of these major peptidergic pathways was confirmed and evidence was obtained for additional, not so prominent, peptidergic projections. The results arae discussed in view of earlier morphological and physiological studies.
Nerve fibres showing immunoreactivity for bombesin have been observed in prevertebral sympathetic ganglia of the rat and guinea-pig. The distribution of bombesin immunoreactive fibres was compared with the distribution of the structurally-related peptide substance P. Dense networks of varicose bombesin immunoreactive fibres were observed, and in the rat coeliac ganglion the density exceeded that of the substance P immunoreactive fibres. Extracts of rat and guinea-pig coeliac ganglion contained material cross-reacting in radioimmunoassays for bombesin tetradecapeptide. Similar concentrations of bombesin and substance P were found in the guinea-pig ganglion, whereas in the rat the concentration of bombesin was about 20 times as high as that of substance P. Two molecular forms of immunoreactive bombesin were identified, neither of which corresponded exactly to synthetic bombesin. The two components were also distinguishable from synthetic substance P. A potential role of a bombesin-like peptide in neural pathways between the gut and prevertebral ganglia is suggested.
In rats treated neonatally with capsaicin there is, in later life, a tendency tendency towards urine retention. Since capsaicin is known to cause irreversible loss of certain primary sensory neurons, notably those containing substance P, we have studied the sensory innervation of the bladder in capsaicin-treated and control rats using retrograde tracing methods and immunohistochemistry; in addition, the motor function of the bladder was assessed in in vitro experiments, using electrical field stimulation. Five days after injection of the fluorescent tracer True Blue into the wall of the bladder, numerous labelled cells were identified in dorsal root ganglia T13, L1, L2, L6, and S1 and smaller numbers of cells were found in T12 and L3. In capsaicin-treated rats the numbers of labelled cells were reduced by over 50% in L1, L6 and S1. In control rats, 10-16% of True Blue labelled cells also contained substance P as demonstrated by indirect immunofluorescence, but in capsaicin-treated rats substance P cells were virtually absent. In in vitro studies, contractions of the detrusor muscle to electrical field stimulation, both before and after atropine, were similar in control and capsaicin-treated rats. We suggest that capsaicin causes urine retention in rats due to an impairment of sensory transmission from the bladder (that could involve substance P) and a consequent failure in the normal micturition reflexes.
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Using the indirect immunohistochemical technique with antisera to cholecystokinin and to substance P, the spinal dorsal horn and dorsal root ganglia of normal and colchicine-treated rats were studied. In the spinal cord a similar distribution of substance P- and cholecystokinin-positive networks in the superficial layers of the dorsal horn was observed. In the dorsal root ganglia several cholecystokinin and substance P immunoreactive cell bodies were seen in colchicine-treated rats. After elution and restaining for substance P, of sections previously stained for cholecystokinin, it was found that all cholecystokinin-positive cells also contained substance P-like immunoreactivity and vice versa.
In rats treated neonatally with capsaicin there were significant reductions in substance P measured by radioimmunoassay in dorsal spinal cord, spinal ganglia, and coeliac ganglia, but concentrations of cholecystokinin octapeptide (CCK8) measured by radioimmunoassay were not decreased. However, immunohistochemical studies using antisera to both substance P and CCK8, respectively, demonstrated decreased immunoreactive material in the dorsal horn of the spinal cord. The nature of the material localized by CCK8 antiserum in immunohistochemistry remains to be resolved.
Substance P is widely distributed in the nervous system. In brain and spinal cord it may act as a transmitter, for example at the central branches of primary sensory neurons. It may also be released from the sensory nerve endings and is thought to be involved in antidromic vasodilatation and in synaptic transmission in autonomic ganglia. In some central neurons substance P is stored together with 5-hydroxytryptamine and thyrotropin-releasing hormone. These neurons project to the ventral horn of the spinal cord, amongst other places. In another system substance P coexists with a cholecystokinin-like peptide. These neurons are localized in the periaqueductal central grey matter and also project to the spinal cord. Finally, injection of a substance P antagonist into the ventral mesencephalon causes marked morphological changes in neurons that contain dopamine, substance P and gamma-aminobutyric acid (GABA).
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By means of the immunohistochemical technique of Coons and collaborators, numerous peptide-containing neurons have been observed in the brain, spinal cord and periphery. These neurons may contain peptides such as substance P, vasoactive intestinal polypeptide (VIP), enkephalin or somatostatin. Some systems are very extensive. For example, immunoreactive substance P has been observed in more than 30 cell groups in the central nervous system, in primary sensory neurons, in sensory neurons in the vagus nerve and in taste buds, and in intestinal neurons. Thus, one and the same peptide can be utilized at many different levels in the nervous system. Several examples are now known where a regulatory peptide occurs together with a classical transmitter, such as a catecholamine, in the same neuron, which suggests the possibility that a neuron can release more than one transmitter substance. Of particular interest is the occurrence of VIP in presumed cholinergic neurons innervating exocrine glands in the cat, and the coexistence of a cholecystokinin (CCK)-like peptide in dopamine neurons projecting mainly to limbic areas. In the former system VIP seems to be responsible mainly for vasodilation, whereas acetylcholine mainly causes secretion. Furthermore, combined infusion of both substances in very low doses results in a marked potentiation of the secretory and vasodilatory responses. Thus, we have an example where two putative transmitters, released from the same nerve endings, seem to cooperate to activate a physiological response (secretion). With regard to the central CCK/dopamine neurons the type of interaction between the two coexisting transmitter candidates is at present unclear. It is suggested that elucidation of different types of coexistence phenomena may advance our understanding of chemical transmission at synapses under normal and pathological conditions, and may lead to new approaches to the treatment of some nervous disorders.
More than 20 peptides have been identified in neurones of the brain, spinal cord and periphery. In several cases a peptide occurs together with a 'classical' transmitter in the same neurone. This raises new questions about the process of neurotransmission and may have important implications for our understanding and treatment of mental disorders.
Tissue pieces of small and large intestine from pre- and postnatal and adult rats, respectively, were transplanted to the anterior eye chamber of the rat. Immunohistochemical studies revealed high numbers of substance P-, vasoactive intestinal polypeptide (VIP)- and enkephalin-containing nerve fibers, especially in the circular smooth muscle layer and in the myenteric plexus of the intestinal tissue transplants. A small number of somatostatin-containing fibers were observed. The distribution patterns of the various peptide-containing neurons in the grafts resembled those which have been described in the intact gastro-intestinal wall. Substance P, VIP, enkephalin and somato-statin immunoreactive fibers were seen also in the host irides. No obvious differences in peptide innervation in grafts and host irides were observed between normal and sympathectomized animals and animals lesioned in the trigeminal ganglion. These findings suggest that most of the peptide-containing fibers in the intestinal transplants and the host irides originate in the transplanted tissue. Thus, it is shown that peptide neurons can survive in this transplantation model, which may be useful for functional studies of intestinal physiology including peptidergic mechanisms.