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X F Zhou

Publications and source records attributed to X F Zhou.

At least 73 records · Page 4Linked to original sources

Peripheral projections of a subpopulation of dorsal root ganglion neurons defined by ovalbumin immunoreactivity.

Previous studies from this laboratory have used antisera to aldehyde-conjugated ovalbumin to localize ovalbumin-like immunoreactivity within a subpopulation of sensory neurons. We have now combined retrograde tracing and immunohistochemical procedures to identify the tissues innervated by sensory neurons which are either immunoreactive or non-immunoreactive for ovalbumin. The fluorescent tracer Di-I was administered to feather follicles, flexor ulnar muscle, subdermis, expansor secundariorum, heart and liver and identified seven days later within corresponding dorsal root ganglia. Most neurons innervating the follicles had large cell somata, and fewer than 3% were immunoreactive for ovalbumin. In contrast, most sensory neurons projecting to subdermis, muscle and expansor secundariorum muscle were of a medium diameter. Approximately 25% of those neurons projecting to the expansor secundariorum, and 60% projecting to the subdermis and muscle, were immunoreactive for ovalbumin. Sensory neurons innervating heart and liver were the smallest, and only 8% were immunoreactive for ovalbumin. The study indicates that sensory neurons innervating different organs have somata with significantly different sizes, suggesting a functional specificity. Moreover, neurons demonstrating either the ovalbumin-IR positive or negative phenotypes show distinct peripheral projections, suggesting that this phenotype may be at least partially controlled by retrograde signals derived from the cells they innervate.

Afferent Pathways↗

An improved procedure for the immunohistochemical localization of nerve growth factor-like immunoreactivity.

Nerve growth factor (NGF) is a survival factor required by a number of neuronal populations including most post-ganglionic sympathetic neurones. NGF has been detected and quantified in many tissues but there is little information regarding its cellular localization. Although it has been argued that histological detection has proven difficult due to the low levels of NGF present, other factors may contribute to prevent its identification. In the present study, we report a method for the histological detection of NGF-like immunoreactivity in the rat superior cervical ganglia (SCG). Adult Wistar-Kyoto rats were perfused briefly with either a high or low pH buffer prior to fixation and routine immunohistochemistry. Polyclonal antibodies to native mouse NGF used in the present study recognized mouse NGF but not recombinant human neurotrophin 3 (rhNT3) or brain-derived neurotrophic factor (rhBDNF) by immunoblot analysis. NGF-like immunoreactivity was localized to most sympathetic neurones. Immunoreactivity was detected in the cytoplasm with dense labelling around nuclei. No stain was seen in sections incubated with normal sheep IgG or from animals perfused with phosphate buffer (pH 7.4) prior to fixation. In addition, axotomy resulted in the disappearance of NGF immunoreactivity which was confirmed by biochemical quantification. Finally, no NGF immunoreactivity was found in neurones of rats treated systemically with NGF antiserum 3 days earlier. Possible mechanisms underlying the improvement of NGF immunohistochemistry by pH manipulation before fixation are discussed.

Animals↗

A subpopulation of chicken primary sensory neurons defined by complete co-localization of peripherin-and ovalbumin-immunoreactivities.

In a previous study, we have demonstrated that an ovalbumin-like antigen is present within approximately one-half of all neurons of chicken spinal ganglia. The current study demonstrates this antigen co-localizes absolutely with neural intermediate filament protein (Peripherin) in small to medium-sized neurons of spinal ganglia. While the function of ovalbumin in neurons is unknown, its precise co-localization with Peripherin suggests a functional role restricted to neurons of a defined phenotype.

Animals↗

Distribution of trkB tyrosine kinase immunoreactivity in the rat central nervous system.

Recent evidence suggests that trkB tyrosine kinase is a high affinity receptor for brain-derived neurotrophic factor (BDNF). BDNF can act as a survival factor for several neuronal subgroups and its mRNA is distributed widely throughout the central nervous system. However, the functional targets of BDNF are poorly defined. We have used immunochemical and immunohistochemical techniques to determine the regional distribution and cellular localization of trkB tyrosine kinase-like immunoreactivity. The staining pattern indicates that the trkB-like antigen is widely distributed and present within both glia and neurons. Astrocytes were the most intensively labelled but many neuronal populations were also stained. In some regions including brain stem, spinal cord, hippocampus and diagonal band of Broca, neurons were stained at varying intensities. In other areas such as the cortex of the forebrain and amygdaloid nucleus, the stain was intense but diffuse, preventing positive identification of the cell types involved. Immunoblot results indicated two separate protein bands in all brain and spinal cord regions examined, of molecular weights 145 and 85 kDa, respectively. These findings aid the definition of neuronal and glial subpopulations of the central nervous system that may utilize BDNF.

Animals↗

Localization of neurotrophin-3-like immunoreactivity in peripheral tissues of the rat.

Neurotrophin-3 (NT-3) mRNA is widely distributed in both the peripheral and central nervous systems but neither the distribution of the native factor nor its physiological function is known. In the present study we produced and characterized an antibody to a synthetic peptide and showed that it specifically recognised endogenous rat and recombinant human NT-3 (rNT-3), but not mouse nerve growth factor and recombinant brain derived-neurotrophic factor. NT-3-like immunoreactivity (NT-3-ir) was detected within the distal tubular cells of the kidney, the zona glomerulosa and reticularis of the adrenal cortex, ganglion cells of the adrenal medulla, red pulp of the spleen, epithelial cells of the intestinal villi and parenchymal cells of the liver. Within peripheral ganglia, NT-3-ir was observed in a subpopulation of large sensory neurons of dorsal root, trigeminal and cochleovestibular ganglia but not in principle neurons of the sympathetic ganglia. These results provide the first evidence for the localization of NT-3-ir and indicate its presence in various peripheral organs and large sensory neurons. We conclude that NT-3 may function outside the nervous system in addition to a neurotrophic role within large sensory neurons.

Animals↗

CGRP immunoreactive neurons in rat dorsal root ganglia do not all contain low-affinity NGF receptor immunoreactivity.

Nerve growth factor (NGF) and its homologues bind to the low affinity NGF receptor (LNGFR). We have determined whether the LNGFR colocalizes with calcitonin gene-related peptide (CGRP) as this peptide is restricted to neurons exhibiting high affinity binding sites for NGF. LNGFR and CGRP immunoreactivities were detected in 57% and 49% of spinal ganglion neurons respectively with different soma areas and only partial colocalization. Our data suggest that the LNGFR may not be essential for a functional neurotrophin response.

Animals↗

Ovalbumin-like immunoreactivity detected in chicken sensory neurons by antibodies to aldehyde-treated ovalbumin.

A method has been developed to raise an antiserum against ovalbumin that can detect this antigen immunohistochemically in chicken sensory ganglia. Ovalbumin-like immunoreactivity has been identified in a subpopulation of chicken dorsal root ganglion neurons by the generation of antibodies to aldehyde-conjugated ovalbumin but not by the antibodies to native ovalbumin, although both antibodies recognize the much higher concentrations of ovalbumin in sections of the oviduct. Biochemical analysis demonstrated that the antigen is more readily detectable in fixed tissue extracts than in fresh tissue extracts. Sensitive immunoblot analysis combined with affinity purification of the antigen, has confirmed that the antigen is of the same molecular weight as ovalbumin. Furthermore, the immunoreactive material elutes at a position identical to native ovalbumin on a molecular sieve column. These findings argue that molecules sensitive to aldehyde fixation may be more readily detected by the use of antisera prepared against aldehyde-modified antigens. The function of the ovalbumin-like antigen in these neurons is unknown.

Animals↗

[Inhibition of photosensitization of morin on DNA synthesis of ascites hepatoma cells].

Morin (0.012, 0.12, 1.2, 12.0 micrograms.ml-1) significantly inhibited the DNA synthesis of ascitic hepatoma (AH) cells. The inhibition of DNA synthesis and cell mortality was dependent on its concentrations as well as the illumination time. Photosensitization of morin was not due to 1O2 and O2-, but closely related to OH. and H2O2. The mechanism of the inhibition may be attributed to the damage of DNA replication template.

Animals↗

Substance P-containing sensory neurons in the rat dorsal root ganglia innervate the adrenal medulla.

The adrenal medulla is innervated by both cholinergic and substance P (SP)-containing fibres via the splanchnic nerve. SP has been shown to modulate catecholamine (CA) secretion in isolated chromaffin cells and in the perfused rat adrenal gland, however, the origin of SP-containing fibres is not known. In the present study, we have combined the techniques of SP immunohistochemistry and retrograde tracing with Fast blue injected into the left adrenal medulla of the rat in order to study whether SP-containing sensory neurons in the dorsal root ganglia innervate the adrenal medulla. The results showed that there were on average 281 +/- 31 SP-like immunoreactive cells in each left dorsal root ganglion, T3-T13 (range, 234 +/- 19 in T4 to 372 +/- 43 in T13, n = 8). The average total number of Fast blue-labelled cells (T3-T13) in 8 experiments was 172 +/- 26, distributed normally about a peak at T8 (33.8 +/- 6.3 cells) and T9 (33.3 +/- 6.8 cells) with the least at T3 (1.5 +/- 0.8) and T13 (5.2 +/- 2.0). No Fast blue-labelled cells were found in the right DRG. In the left DRG, the average number of cells exhibiting both SP and Fast blue labelled cells were distributed from T7 to T9. These results demonstrate that SP-containing sensory neurons in the dorsal root ganglia provide an ipsilateral innervation of the adrenal medulla in rats.

Adrenal Medulla↗

Substance P modulates the time course of nicotinic but not muscarinic catecholamine secretion from perfused adrenal glands of rat.

1. Substance P (SP) and acetylcholine (ACh) are contained within the splanchnic nerve terminals in the adrenal gland and can be released in response to stress. In the rat, the release of aCh brings about secretion of catecholamines (CA) by acting on nicotinic and muscarinic receptors on the adrenal chromaffin cells. 2. In the present study, we have used a rat isolated adrenal gland preparation to investigate the effects of SP, perfused at different concentrations, on CA secretion evoked by 10(-5) M nicotine and 10(-4) M muscarine. 3. In the first 10 min stimulation period (S1), in the absence of SP, nicotine (10(-5) M) evoked substantial and equal secretion of noradrenaline (NA) and adrenaline (Ad). In a second 10 min stimulation period (S2), carried out 18 min after S1, the nicotinic response was desensitized. In contrast, the muscarinic response, which preferentially evoked Ad secretion in S1 (Ad/NA: 8.7/1), was well maintained in S2. 4. SP present in S1 had no effect on desensitization of the subsequent nicotinic response in S2. 5. At low concentrations (10(-7)-10(-10) M), SP changed the time course of nicotine-induced CA secretion during S1 by enhancing CA secretion in the first 4 min and inhibiting CA secretion thereafter. The maximal effect occurred at 10(-9) M SP. 6. At a higher concentration (10(-5) M), SP inhibited total nicotinic CA secretion throughout S1 and produced a biphasic secretion of CA (depressed in the presence of SP and enhanced after wash out of SP). Pre-exposure of adrenal glands to SP (10-' to 10- M) for 10min produced marked inhibition of the nicotine-induced CA secretion. 7. In contrast to the effect of SP on the nicotinic response, SP from 10- to 10-SM had no effect on muscarinic CA secretion. 8. This difference in sensitivity of the nicotinic and muscarinic responses to SP points to a diversity of mechanisms available for control of adrenal catecholamine secretion. In addition to the ability of SP to increase or decrease the total amount of adrenal CA secretion, dependent on the concentration of SP, the present study shows that SP can change the time-course of nicotinic CA secretion. These results with the rat adrenal gland perfused in vitro suggests both a quantitative and temporal role for SP as a novel modulator of adrenal CA secretion.

Adrenal Glands↗

Capsaicin-sensitive sensory neurons are involved in the plasma catecholamine response of rats to selective stressors.

1. The effect of capsaicin pre-treatment on adrenal catecholamine (CA) secretion in response to stress is controversial. In earlier experiments performed under pentobarbitone anaesthesia, the release of CA in response to stress was complicated by the effects of the barbiturate anaesthesia. 2. In the present study we have used conscious freely moving rats with indwelling cannulae to study the effect of neonatal capsaicin pre-treatment on the plasma CA response to different types of stressors (swimming stress, hypovolaemic stress, immobilization stress and cold stress). 3. After swimming for 20 min, plasma noradrenaline (NA) levels increased by 8-fold and adrenaline by 2-fold in control rats. The increase in plasma NA levels in the capsaicin group was attenuated at 10 min of swimming compared with the vehicle group (P < 0.05). 4. With hypovolaemic stress, there were no differences in plasma CA levels, blood pressure and heart rate between the capsaicin group and the vehicle group. There were also no differences in plasma CA levels after immobilization stress between the two groups. 5. With cold stress, plasma NA levels increased 5-fold and adrenaline levels by 3-fold over basal at 45 min in the vehicle pre-treated rats. This increase was not observed in the capsaicin group. 6. Immunoreactive substance P was depleted by only 68% in the splanchnic nerve following capsaicin pre-treatment. If the remaining 32% was biologically active substance P then it could account for the maintenance of the response to hypovolaemic and immobilization stress. However, it might be possible that the responses to hypovolaemic and immobilization stresses could be attenuated if a more complete depletion were achieved. 7. These results in conscious rats indicate that capsaicin-sensitive sensory neurons are required for plasma CA response to selective stressors. They are required for CA output in response to cold stress and to the early phase of swimming stress, but not to hypovolaemic stress and immobilization stress.

Animals↗

Role of capsaicin-sensitive neurons in catecholamine secretion from rat adrenal glands.

Sensory fibres innervate the adrenal medulla but their function is not known. In the present study, we have used the sensory neurotoxin capsaicin to evaluate the effect of capsaicin sensitive sensory fibres on catecholamine (CA) secretion from isolated perfused rat adrenal glands. CA secretion in response to 1 and 10 min electrical field stimulation of adrenal nerve terminals was significantly attenuated in the adrenal glands of adult rats pretreated as neonates with capsaicin and was frequency dependent, being more pronounced at the higher frequencies of stimulation (5 to 30 Hz) than at the low (0.3, 1 Hz) frequencies. Perfusion of control rat adrenal glands with capsaicin did not evoke CA secretion, but did increase CA secretion in response to perfusion with nicotine. Perfusion with capsaicin for 30 min (but not for 4 min) reduced the CA secretory response to subsequent nerve stimulation. The results suggest that capsaicin sensitive sensory neurons innervating the adrenal medulla are involved in the regulation of adrenal CA secretion evoked by electrical stimulation of adrenal nerve terminals.

Adrenal Glands↗

Hypovolaemia can potentiate hypoglycaemic stress-induced adrenaline release in the anaesthetized rat.

The interaction between hypoglycaemic stress and hypovolaemic stress in promoting adrenaline secretion from the adrenal medulla has been studied in anaesthetised and conscious rats. In anaesthetized rats, insulin (1 IU/kg, i.v.) markedly increased plasma adrenaline concentrations whereas blood sampling alone did not. The hypoglycaemic response to insulin was greatly reduced if donor blood was used instead of saline to replace the withdrawn blood. The hypoglycaemic response was abolished by captopril or saralasin. The results suggest that adrenaline secretion in response to hypoglycaemic stress in anaesthetized rats is potentiated by hypovolaemic activation of the renin-angiotensin system. In contrast, in fully conscious rats, the response to hypoglycaemia was not abolished by captopril, indicating that in the absence of barbiturate anaesthesia, the hypoglycaemic release of catecholamines is not potentiated by the renin-angiotensin system.

Adrenal Medulla↗

Peptide regulation of adrenal medullary function.

The discovery of peptides in the splanchnic nerve and adrenal gland, and their co-existence with conventional neurotransmitters raises questions about their possible functional roles in catecholamine (CA) secretion and gene transcription in the adrenal gland. Short-term, stress-induced CA secretion is regulated biphasically by substance P (SP) which inhibits acetylcholine (ACh) action at SP greater than 10(-6) M and facilitates CA secretion in response to metabolic and physical stressors, ACh or electrical stimulation at SP less than 10(-6) M. Long-term, gene transcription of phenylethanolamine-N-methyltransferase (PNMT) is exerted by glucocorticoids, and gene transcription of Proenkephalin-A by agents such as histamine, angiotensin II and VIP that increase cyclic AMP (cAMP). The final products of these two genes, adrenaline and Met-enkephalin and congeners, are co-stored in chromaffin granules of adrenaline cells but gene expression of these products is clearly under differential control.

Adrenal Medulla↗

Substance P has biphasic effects on catecholamine secretion evoked by electrical stimulation of perfused rat adrenal glands in vitro.

The adrenal medulla is innervated by the splanchnic nerve which contains substance P-immunoreactive fibres originating in the dorsal root ganglia but whose function in the adrenal medulla is not known. In the present study, we have examined the effect of exogenous substance P on catecholamine secretion and [3H]-choline overflow evoked by field stimulation of the perfused rat adrenal gland. Substance P had biphasic effects on catecholamine secretion evoked by field stimulation: at the lower concentrations (10(-7)-3 x 10(-6) M), substance P facilitated catecholamine secretion whereas at higher concentrations (3 x 10(-5) M), substance P inhibited catecholamine secretion. The effects of substance P were long-lasting: substance P at all concentrations present during the second of three 6-min stimulations increased catecholamine secretion during the third stimulation. The enhancement of catecholamine secretion by substance P during field stimulation was not blocked by atropine but was substantially blocked by hexamethonium plus atropine. Substance P at all concentrations inhibited the evoked [3H]-choline overflow with a maximal inhibition of 60% (at 10(-6) M). In the absence of stimulation, substance P (10(-7)-3 x 10(-5) M) had no effect on the basal catecholamine secretion or basal [3H]-choline overflow. These results indicate that substance P has modulatory effects on adrenal catecholamine secretion and [3H]-choline overflow evoked by electrical stimulation. Presynaptically, substance P inhibits transmitter overflow from cholinergic nerve terminals and post-synaptically substance P has biphasic effects on catecholamine secretion dependent on the substance P concentration. These results suggest a functional role for substance P-containing fibres innervating the adrenal medulla.

Adrenal Glands↗

Effect of capsaicin-sensitive sensory nerves on plasma glucose and catecholamine levels during 2-deoxyglucose-induced stress in conscious rats.

1. Sensory fibres innervate the adrenal medulla but their function is not known. In this paper we have studied the effect of capsaicin-sensitive sensory fibres on the adrenal catecholamine (CA) response and blood glucose response to 2-deoxyglucose (2-DG)-induced glucopenic stress in conscious rats. 2. 2-DG at 500 mg kg-1 (i.v.) induced a 2.5 fold increase in plasma glucose levels, a 3.5 fold increase in inferior vena caval (i.v.c.) plasma noradrenaline (NA) levels and a 7 fold increase in i.v.c. plasma adrenaline (Ad) levels over 60 min. The hyperglyaemia in response to 2-DG was attenuated by pentolinium and by left splanchnicotomy plus right adrenalectomy. These procedures also caused a complete inhibition of the increase in plasma CA. 3. The hyperglycaemia in response to 2-DG was attenuated by pretreatment of rats with capsaicin as neonates, suggesting that capsaicin-sensitive sensory fibres are required for regulation of plasma glucose in response to glucopenic stress. 4. The increase in i.v.c. plasma CA levels in response to 2-DG during the early phase of glucopenia (first 30 min) in the conscious rats pretreated with capsaicin was the same as in the rats pretreated with vehicle alone. During the later phase of glucopenia (after 45 min), the increase in plasma CA levels in rats pretreated with capsaicin was higher than in the rats pretreated with vehicle alone. 5. In vehicle-pretreated rats and capsaicin-pretreated rats the tissue NA and Ad levels in the adrenal medulla after 8 h of stress were depleted to the same extent. However, tissue CA levels in the capsaicin group recovered faster over 24 h than in the vehicle group. 6. These results indicate that capsaicin-sensitive sensory fibres are not required to maintain adrenal CA secretion during glucopenic stress in the conscious rat but are required for maintenance of blood glucose levels.

Adrenal Medulla↗

Substance P increases catecholamine secretion from perfused rat adrenal glands evoked by prolonged field stimulation.

1. We have studied the effect of substance P (SP) on catecholamine (CA) secretion evoked by prolonged field stimulation from the perfused rat adrenal gland in vitro. 2. Previous studies show that SP has an inhibitory effect on the nicotinic response in a number of different tissue preparations. In the present study, we found that SP at concentrations from 10(-7) to 10(-6) M markedly increased CA secretion evoked by prolonged high-frequency field stimulation but not that evoked by low-frequency field stimulation. 3. In the absence of field stimulation SP by itself had no direct effect on CA secretion. This indicates that SP acts as a neuromodulator rather than a neurotransmitter to increase CA secretion from the adrenal medulla. 4. The nicotinic receptors were still functional following a period of prolonged field stimulation (120 min) because CA secretion was markedly increased in response to the nicotinic agonist dimethylphenylpiperazinium (DMPP). 5. Capsaicin pre-treatment reduced CA secretion evoked by prolonged field stimulation and the facilitatory action of SP on CA secretion lasted longer in these capsaicin-pre-treated rats than in controls, indicating that SP-containing capsaicin-sensitive fibres innervating the adrenal medulla may be involved in the regulation of CA secretion. 6. In parallel with the increase in CA secretion, 3H overflow from the splanchnic nerve pre-labelled with [3H]choline was also increased by SP. The increase in CA secretion by SP lasted longer than the increase in 3H overflow. These results suggest that SP facilitates CA secretion from the adrenal gland at two levels: (1) pre-synaptically by facilitating ACh release from splanchnic nerve terminals, and (2) post-synaptically by modulating the nicotinic secretory response by protection against nicotinic desensitization of secretion. 7. The present studies provide further evidence that endogenous SP in the splanchnic nerve may modulate CA secretion during stress.

Adrenal Glands↗