H. Päivärinta

[Met5]enkephalin-Arg6-Phe7- and [Met5]enkephalin-Arg6-Gly7-Leu8-immunoreactive nerve fibres and neurons in the superior cervical ganglion of the rat

[Met5]enkephalin-Arg6-Phe7-(MEAP-) and [Met5]enkephalin-Arg6-Gly7-Leu8-(MEAGL-) immunoreactivity was studied by indirect immunohistochemistry in the superior cervical ganglion of the rat with specific antisera produced in rabbits against the corresponding synthetic opioid peptides. Several MEAP- and a few MEAGL-immunoreactive principal nerve cells were observed in the ganglion, while the small intensely fluorescent cells appeared as non-reactive. The superior cervical ganglion also contained dense networks of MEAP- and MEAGL-immunoreactive nerve fibres, which often formed basket-like structures around the principal nerve cells and small intensely fluorescent cells. After ligation of the preganglionic nerve trunk with simultaneous transection of the main postganglionic trunks, a distinct accumulation of both MEAP- and MEAGL-immunoreactivity was observed on both sides of the ligature. Ligation of the preganglionic nerve trunk caused a marked decrease in the number of both MEAP- and MEAGL-immunoreactive nerve fibres in the ganglion. Ligation of the main postganglionic nerve trunks with simultaneous preganglionic nerve division resulted in accumulation of MEAP- and MEAGL-immunoreactive material on the ganglionic side of the ligature in both the external and internal carotid nerve. After division of both the pre- and postganglionic nerve trunks, some immunoreactive nerve fibres and principal nerve cells were still observed in the ganglion. A few immunoreactive neurons and nerve fibres were also observed in the ganglion stellatum. A large number of MEAP- and MEAGL-immunoreactive nerve fibres was detected in the spinal cord at the levels C6-Th6. A few neurons in the intermediolateral cell column of the spinal cord at levels C8-Th1 showed MEAP- but not MEAGL-immunoreactivity. The cultured superior cervical ganglion contained a few MEAP-immunoreactive neurons, and the fibre outgrowth showed immunoreactivity both to MEAP and MEAGL. In electron microscopy, MEAGL-immunoreactivity in the superior cervical ganglion was localized in nerve fibres containing neurotubules and in principal nerve cells. The present results demonstrate that the rat superior cervical ganglion contains both extrinsic and intrinsic MEAP- and MEAGL-immunoreactive nerve fibres. Most of these fibres are of preganglionic origin. Both the principal nerve and small intensely fluorescent cells are often surrounded by MEAP- or MEAGL-immunoreactive nerve fibres and may receive innervation by these fibres. Several ganglionic neurons projecting to the sympathetic target tissues show MEAP- and/or MEAGL-immunoreactivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Substance P-like immunoreactivity in the rabbit inner ear

The auditory and vestibular sense-organs of the rabbit were examined for the presence of substance P(SP) by an immunohistochemical technique using a monoclonal SP-antibody. Between 30 and 40 per cent of the cells of the vestibular ganglia and about 50 per cent of the spiral ganglion cells innervating the extreme basal part of the cochlea showed SP-like immunoreactivity. The neural elements in the vestibular sensory epithelia, notably the calyx-formed nerve terminals of type I sensory cells of the maculae, also showed strong SP-like immunoreactivity. The findings suggest the possibility of a modulator or transmitter role for SP in the inner ear function.

Pre- and postnatal development of 5-hydroxytryptamine-immunoreactive cells in the superior cervical ganglion of the rat

The pre- and postnatal development of 5-hydroxytryptamine (5-HT)-immunoreactive cells in the superior cervical ganglion of the rat was studied by indirect immunofluorescence method with an antibody to a conjugate of bovine serum albumin and 5-HT. The superior cervical ganglia from 11-day-old embryos to 90-day-old postnatal rats were examined. 5-HT-immunoreactive cells were first detected in the superior cervical ganglion on the 12th day of gestation. At this stage of development the ganglionic cells formed a condensed group, and most of them showed 5-HT-immunoreactivity. During later prenatal development the relative number of the 5-HT-immunoreactive cells in the ganglion decreased and most ganglionic cells appeared as non-reactive. 5-HT-immunoreactive cells showed some variation in size and fluorescence intensity during the whole prenatal development. In the ganglia of newborn rats a wide range of 5-HT-immunoreactive cell sizes was detected. Two types of 5-HT-immunoreactive cells were distinguished in the ganglia of 7- to 35-day-old rats: small (5-15 microns in diameter) cells and large (15-30 microns in diameter) cells, resembling in size small intensely fluorescent (SIF) cells and principal nerve (PN) cells, respectively. The relative number of the large 5-HT-immunoreactive cells gradually decreased after the first postnatal week, and these cells were not detected in the ganglia of 90-day-old rats. Small 5-HT-immunoreactive cells constantly formed clusters in the ganglia of postnatal rats. A marked, statistically significant increase in the density of 5-HT-immunoreactive cells occurred during the 4th postnatal week. The 5-HT-immunoreactive cells appeared in the prenatal superior cervical ganglion of the rat approximately at the same time as the cells showing formaldehyde-induced catecholamine fluorescence. Transient occurrence of the large 5-HT-immunoreactive cells during the first postnatal weeks of development indicates, that in the superior cervical ganglion of the rat, 5-hydroxytryptamine may be expressed in some developing PN cells early postnatally. After the 5th postnatal week, the large 5-HT-immunoreactive neuronal cells cannot be detected, and 5-HT is expressed only in the SIF cells.

Localization of l-glutamate decarboxylase and GABA transaminase immunoreactivity in the sympathetic ganglia of the rat

The location of L-glutamate decarboxylase and gamma-aminobutyrate (GABA)-transaminase immunoreactivity in the superior cervical ganglion and in the coeliac-superior mesenteric ganglion complex of the rat was studied by an indirect immunofluorescence method and by immunoelectron microscopy, with specific antisera raised in rabbits against the corresponding enzymes. In light microscopy, several glutamate decarboxylase- or GABA-transaminase-immunoreactive principal nerve cells were detected in the superior cervical ganglion and coeliac-superior mesenteric ganglion complex. In addition, numerous small cells in both the superior cervical ganglion and coeliac-superior mesenteric ganglion complex showed intense immunoreactivity to glutamate decarboxylase or GABA-transaminase. The small cells were 10-20 micron in diameter and resembled in size and morphology the small intensely fluorescent cells. In consecutive sections, the small glutamate decarboxylase-immunoreactive cell clusters also showed immunoreactivity to tyrosine hydroxylase, suggesting that these cells contain the enzymes for both GABA and catecholamine synthesis. In the superior cervical ganglion and in the coeliac-superior mesenteric ganglion complex, GABA-transaminase immunoreactivity was also localized in fibre-like processes around and between the principal nerve cells, in nerve trunks traversing the ganglia, and around or in close contact with ganglionic blood vessels. Furthermore, GABA-transaminase immunoreactivity was observed in fibre-like structures close to the capsule of the ganglia. Division of the preganglionic nerve trunk of the superior cervical ganglion caused no detectable change in GABA-transaminase immunoreactivity in the ganglion. In immunoelectron microscopy of the superior cervical ganglion, GABA-transaminase immunoreactivity was localized in nerve fibres in association with neurotubules. A large number of GABA-transaminase labelled principal nerve cells were detected, containing immunoreactivity evenly distributed in their cytoplasm. GABA-transaminase immunoreactivity was also observed in satellite cells and their processes in the superior cervical ganglion. The present immunocytochemical results provide evidence that the rat sympathetic ganglia contain an intrinsic neuronal system showing histochemical markers for GABA synthesis and inactivation, but its functional role in the modulation of ganglionic neurotransmission remains to be established.

Histamine-immunoreactive cells in the superior cervical ganglion and in the coeliac-superior mesenteric ganglion complex of the rat.

SummaryHistamine-immunoreactive small cells were detected in the superior cervical ganglion and in the coeliac-superior mesenteric ganglion complex of the rat with a specific antiserum produced in rabbits. Most histamine-immunoreactive cells were arranged in clusters, often around small blood vessels. Solitary immunoreactive small cells were also observed and they were easily distinguished from mast cells. The principal nerve cells showed no immunore-activity to histamine and no histamine-containing nerve fibers were detected in the ganglia by the present method. Due to close morphological similarities, it is concluded that the small immunoreactive cells observed in the present study represent small intensely fluorescent (SIF) cells first detected by formaldehyde-induced catecholamine fluorescence.

Histamine-immunoreactive endocrine cells in the adrenal medulla of the rat

Histamine is widely distributed in various mammalian tissues and it has been shown that histamine is located in mast cells as well as in other structures. Biochemical evidence has been presented that histamine acts as a neurotransmitter in the central nervous system. Immunohistochemical studies have demonstrated the location of histamine-immunoreactive neuronal cells in both the central and peripheral nervous system. Biochemical studies have shown that histamine is present in the adrenal gland, while the location of histamine in the adrenal medulla is not known. There is pharmacological and biochemical evidence that exogenous histamine affects the catecholamine secretion of the adrenal medulla. The present study was undertaken to examine the location of histamine in the rat adrenal medulla by an indirect immunofluorescence method using a specific histamine antiserum. We now report the presence of histamine-immunoreactive endocrine cells in the adrenal medulla of the rat and suggest that histamine is located in the noradrenaline-secreting cells.

Is Substance P the Neurotransmitter in the Vestibular End Organs

The vestibular end organs of the rabbit were fixed intracardially with phosphate buffered 4% formaldehyde, sectioned for 15 micron cryostat sections and incubated with monoclonal substance P (SP) antibody. Specific SP-like immunoreactivity (SPLI) was observed within and below the sensory epithelia of both utricular and saccular maculae and ampullary cristae. Particularly strong SPLI was seen in the maculae which showed many SPLI-positive axons in the subepithelial space and a network of SPLI-positive structures in the basal zones of the sensory epithelium. The calyceal terminals of type I hair cells showed regularly SPLI. The possibility of a transmitter or modulator role for SP in the vestibular function is suggested.

Phenylethanolamine-N-methyltransferase-immunoreactive cells in developing rat superior cervical ganglion and the effect of hydrocortisone on their number

Rats were subcutaneously injected with hydrocortisone acetate. Control rats were similarly injected with saline. PNMT‐immunoreactive cells in the superior cervical ganglia were immunohistochemically demonstrated, and the cell densities (number/mm3) of the PNMT‐immunoreactive cells were counted.

Effect of hydrocortisone on the number of small intensely fluorescent cells in the rat superior cervical ganglion during pre- and postnatal development.

During the first postnatal week hydrocortisone causes a massive increase in the number of small intensely fluorescent (SIF) cells. The purpose of the present study was to investigate whether the number of SIF cells can be increased with hydrocortisone also prenatally and after the first postnatal week.

Neural control of the middle ear mucosa.

Neurogenic inflammation is assumed to be an important local defence mechanism against noxious stimuli in various parts of the body. Recent experimental evidence suggests substance P(SP) to be the mediator of this mechanism in the respiratory tract. Immunohistochemically, SP has earlier been reported to be present in the middle ear mucosa of the cat and guinea pig. Our earlier study demonstrated SP-like immunoreactive (SPLI) nerve fibres in the middle ear mucosa and tympanic nerve of the rat, guinea pig and man. This study shows that in experimental animals, many neuron cell bodies of the sensory ganglia of the glossopharyngeal nerve show SPLI. It is suggested that SP may be involved in the neurogenic inflammatory reaction leading to the mucosal middle ear disease.