Successful bone marrow harvest during pregnancy.
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Adult↗
Biomedical subjects
R H Ho
Publications and source records attributed to R H Ho.
Evidence for a synergistic effect of the HIV-1 envelope protein gp120 and brain-derived neurotrophic factor (BDNF) leading to enhanced expression of somatostatin neurons in aggregate cultures derived from the human fetal cortex.
Changes in the expression of somatostatin (SRIF) have been observed in the brains of HIV encephalitis. Since gp120 is thought to play a major role in AIDS-associated abnormalities in the brain, we addressed the question: Does gp120 alter the functional expression of human fetal SRIF neurons in culture and if so, is this effect fetal-age dependent? Aggregate cultures, obtained from cortices of nine fetuses (14.9-20.7 weeks), were exposed for 7 days to BDNF or BDNF+gp120; BDNF induced production of SRIF during the subsequent 24-48 h was assessed. Similar effects of BDNF and gp120 were observed in the 9 brain-cultures. A 7-day exposure to BDNF alone led to a significant increase in SRIF production (p=0.014), whereas exposure to gp120 alone did not. Co-exposure to BDNF and gp120 led to an increase in BDNF-induced SRIF production which was significantly greater than that after BDNF alone (p=0.006). These effects were BDNF- and gp120-dose dependent and they were not accompanied by changes in DNA content of the aggregates nor in lactate dehydrogenase activity in the medium; indicating that gp120 did not lead to a major loss of cell integrity. These results are consistent with a synergistic effect of BDNF and gp120 leading to enhanced functional expression of the signalling pathway(s) mediating BDNF induction of SRIF production; an effect expressed by fetal brains throughout the 2nd trimester of gestation. Thus, this culture system can serve as a model to study the mechanism(s) underlying the early interactions between gp120 BDNF in the developing human brain.
Brain-Derived Neurotrophic Factor↗
Opposite effects of astrocyte-derived soluble factor(s) on the functional expression of fetal peptidergic neurons in aggregate cultures: enhancement of neuropeptide Y and suppression of somatostatin.
Previous studies established that fetal rat and human neuropeptide Y (NPY) cortical neurons in aggregate cultures are differentially regulated. Whereas brain-derived neurotrophic factor (BDNF) or phorbol 12-myristate-13-acetate (PMA) induces NPY production in rat cultures, only PMA does so in human cultures. We addressed these questions: 1) Do soluble products of rat or human astrocytes (conditioned medium; rCM and hCM, respectively) enhance the functional expression of cultured NPY neurons and if so, do they enhance the expression of somatostatin (SRIF) neurons as well? 2) Is the NPY-enhancing activity (EA) in the CM species specific? rCM enhanced (approximately 2-fold) both basal and BDNF-stimulated production of NPY and coculture of rat aggregates and astrocytes did not prevent this NPY-EA. Likewise, the hCM enhanced (approximately 2.5-fold) basal and PMA-stimulated production of NPY by human aggregates. Moreover, the hCM enhanced NPY production by rat aggregates and rCM enhanced NPY production by human aggregates. In addition, rCM and hCM each enhanced BDNF-, forskolin-, or PMA-stimulated NPY production by rat aggregates. Under each of the above conditions, the rCM/hCM suppressed (approximately 50%) production of SRIF by rat aggregates. In summary, secretory products of rat and human astrocytes exert opposite effects on the functional expression of NPY and SRIF neurons in culture: enhancement of NPY and suppression of SRIF. By the criteria evaluated in this study, these astrocyte-derived activities do not exhibit species specificity.
Animals↗
Differential potencies of cocaine and its metabolites, cocaethylene and benzoylecgonine, in suppressing the functional expression of somatostatin and neuropeptide Y producing neurons in cultures of fetal cortical cells.
Using aggregate cultures derived from 17-day-old fetal rat cortex, we addressed the question: Does cocaine alter the functional expression of neuropeptide Y (NPY) and somatostatin (SRIF) neurons and, if so, are cocaethylene (CE) and benzoylecgonine (BZE) as active as cocaine? NPY/SRIF production in response to brain-derived neurotrophic factor (BDNF) or phorbol-12-myristate-13-acetate (PMA) was used as a functional criterion. A 5-day exposure to cocaine did not affect basal or stimulated (BDNF or PMA) production of NPY but it markedly suppressed BDNF- or PMA-stimulated production of SRIF. Exposure to CE led to a drastic suppression of basal as well as stimulated (BDNF or PMA) production of both NPY and SRIF. These effects of cocaine and CE were concentration dependent (1-100 microM). BZE did not alter any of these functional parameters. Next, we evaluated the fate of cocaine, CE, and BZE in the culture medium. Cocaine was converted to BZE, whereas BZE was not converted to cocaine. CE was converted to cocaine and BZE, with substantial amounts of cocaine and CE remaining in the medium after 72 hr (approximately 20% each). In summary, cocaine, CE, and BZE exhibited differential potencies in suppressing the expression of cultured NPY and SRIF neurons: CE was more potent than cocaine and BZE was inactive. SRIF neurons were more susceptible than NPY neurons to the effects of cocaine. The higher potency of CE may be due to a property of the compound and/or to CE serving as a source for a slow, continuous formation of cocaine by the brain cells themselves.
Animals↗
Evidence for GAP-43 within descending spinal axons in the North American opossum, Didelphis virginiana.
We have shown previously that GAP-43, a growth associated protein characteristically present in growing and regenerating axons, is relatively abundant in the spinal cord of adult opossums. In the present study, we combined the orthograde transport of the fluorescent marker Fluoro-Ruby with immunofluorescence for GAP-43 to determine if any of it is present within descending spinal axons. When Fluoro-Ruby was injected into the red nucleus and midbrain tegmentum, the medial pontine or medullary reticular formation, the medullary raphe or the lateral vestibular nucleus, axons were labeled in the expected areas of the spinal cord, but in most cases none showed evidence for GAP-43. In two of the four cases with rubral injections, however, a few labeled axons within the rubrospinal tract showed GAP-43 immunofluorescence, and in one case with an injection of the gigantocellular reticular nucleus and adjacent raphe, labeled axons within lamina IX immunostained for the protein. Since serotoninergic neurons are present within the gigantocellular reticular nucleus and adjacent raphe, and axons of the same phenotype are abundant within lamina IX, we asked whether serotoninergic axons contain GAP-43. When sections of the spinal cord were immunostained for both serotonin and GAP-43, many axons within lamina IX showed evidence for both substances. Such axons appeared to contact presumptive motoneurons. In cases with Fluoro-Ruby injections of the forelimb motor cortex, labeled axons were present within the pyramidal tract, and some of them showed evidence for GAP-43.
Animals↗
Pre-pro-somatostatin mRNA in the developing rat spinal cord with special reference to ventral horn motoneurons.
The expression of pre-pro-somatostatin (ppSOM) mRNA in the spinal cord of the developing rat was determined by in situ hybridization. Expression of ppSOM mRNA was detected in cells in the gray matter of the dorsal and ventral horns at day E15, the earliest stage examined in this study. Expression of ppSOM mRNA persisted during development and into adulthood in the dorsal horn. In contrast, ppSOM mRNA expression in presumptive ventral horn motoneurons was apparent during pre- and perinatal periods, but it was not observed in adult rats. These findings are consistent with the hypothesis that ppSOM is transiently expressed by motoneurons and that a peptide derived from it may serve a role in the development of the neuromuscular junction.
Aging↗
Golgi impregnated somatostatin immunoreactive neurons in lamina II of the rat spinal cord.
The morphology of somatostatin immunoreactive (SOM-I) neurons in lamina (L) II of the rat spinal cord was determined using a combination of Golgi impregnation and immunohistochemistry. Golgi-impregnated SOM-I neurons that resembled islet, stalked and other cells were observed. Islet cells are considered to be inhibitory interneurons while stalked cells are excitatory and are thought to relay information from primary afferent neurons to L I projection cells. The heterogeneous morphology of SOM-I neurons suggest they have diverse functions.
Animals↗
Evidence for dorsal root projection to somatostatin-immunoreactive structures in laminae I-II of the spinal dorsal horn.
In order to determine if somatostatin (SOM)-immunoreactive (I) cell bodies and processes in lamina (L) II of the rat spinal cord receive dorsal root input, the latter were anterogradely labeled by wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). SOM-I structures were demonstrated by immunohistochemistry. Cell bodies labeled transscellularly or transsynaptically by WGA-HRP and immunohistochemically for SOM were present in L II. In addition, a L I cell was double labeled. These results suggest that some dorsal root axons innervate SOM-I neurons in L I-II of the spinal cord. In addition to confirming immunohistochemical observations in published reports, we have revealed SOM-I central terminals in the type II glomerulus. Further, a SOM-I CI-terminal, presumed to be of primary afferent origin, contacted a SOM-I dendrite in L II. Since SOM has been implicated in nociceptive function in the dorsal horn, it is possible that some of the SOM-I structures identified are involved in nociceptive processing.
Animals↗
Gibberellin A(4/7) promotes cone production on potted grafts of eastern white pine.
Potted grafts of eastern white pine (Pinus strobus L.) growing in a heated greenhouse were sprayed weekly with 0, 250 or 500 mg l(-1) of gibberellin A(4/7) for four different 6-week periods between mid-May and mid-September. At different periods, gibberellin A(4/7) concentrations of 250 and 500 mg l(-1) were effective in promoting pollen- and seed-cone production. Spraying in May and June, during the period of rapid terminal shoot elongation, significantly increased pollen-cone production, whereas spraying in August and September, about a month after terminal shoot elongation, significantly increased seed-cone production.
Journal Article↗
Phialophora mutabilis keratomycosis.
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Adult↗
Organization of midbrain catecholamine-containing nuclei and their projections to the striatum in the North American opossum, Didelphis virginiana.
Presumptive catecholamine (CA) neurons in the opossum midbrain were identified by tyrosine hydroxylase immunohistochemistry. In the midline, small to moderate number of CA cells were present in the rostral third of the nucleus raphe dorsalis and throughout the nucleus linearis. Ventrolaterally, such cells were observed in the deep tegmental reticular formation, in all subnuclei of the ventral tegmental area, and in the three subdivisions of the substantia nigra. The CA cells in these areas conform to the dopamine cell groups, A8, A9, and A10 as described in the rat. In several areas there appeared to be no separation between the CA neurons belonging to cytoarchitecturally different nuclei. In order to determine which CA neurons gave rise to striatal projections, the neostriatum was injected with True Blue (TB), and sections through the midbrain were processed for tyrosine hydroxylase (TH) and visualized by immunofluorescence. Neurons containing both TB and TH were observed in each of the CA cell groups mentioned above. The distribution of these cells confirmed organizational features that may be unique to the opossum's substantia nigra. In addition, different patterns of labeling resulted from caudate versus putamen injections, suggesting a rudimentary medial to lateral topography in the organization of nigrostriatal projections. Although our results suggest that the organization of midbrain CA neurons in the opossum is similar to that in placental mammals, it is clear that differences exist.
Animals↗
The origin of serotoninergic projections to the lumbosacral spinal cord at different stages of development in the North American opossum.
We have employed immunohistochemistry and the retrograde transport of Fast blue to study the origin of serotoninergic projections to the lumbosacral spinal cord at different stages of development in the North American opossum. A few serotoninergic axons are present in the lumbosacral cord at birth, 12 days after conception, and serotoninergic neurons are numerous in the brainstem where they are present in most, if not all, of the areas which contain them in the adult animals. A few neurons of the caudal raphe and adjacent reticular formation were labeled by lumbar injections of Fast blue on postnatal day 1, and by postnatal day 3, labeled neurons were numerous within all areas which provide serotoninergic projections to the lumbosacral cord in adult animals. By postnatal day 11, it was possible to combine Fast blue labeling with immunofluorescence to show that some of the labeled neurons were serotoninergic. By postnatal day 24, neurons which provide serotoninergic projections to the lumbosacral cord were especially numerous and some of them were found in areas which do not provide comparable projections in adult animals. In developing and adult animals, few, if any, neurons were labeled in the dorsal raphe or superior central nuclei. We have shown previously that serotoninergic axons do not innervate laminae I and II of the lumbosacral cord until approximately postnatal day 50, although they are present in the marginal zone at birth and have grown into laminae III-X by postnatal day 15. Since serotoninergic axons which project to laminae I and II originate within the raphe magnus and adjacent reticular formation, and those areas provide serotoninergic projections to the spinal cord well before postnatal day 50, it is possible that serotoninergic innervation of laminae I and II is provided by late growth of collaterals from axons that have been present in the marginal zone for some time.
Amidines↗
The origins of supraspinal projections to lumbosacral and cervical levels of the spinal cord in the gray short-tailed Brazilian opossum, Monodelphis domestica.
Retrograde tracing techniques were used to identify supraspinal neurons that project to sacral, lumbar and cervical levels of the spinal cord in the gray short-tailed Brazilian opossum, Monodelphis domestica. Injections of Fast Blue, True Blue or wheat germ agglutinin conjugated to horseradish peroxidase into the sacral or lumbar cord labeled neurons in hypothalamic and brainstem nuclei reported to innervate the same levels in other mammals. Injections at cervical levels produced extensive labeling in the same areas as well as labeling within the isocortex, the medial preoptic area, the central and basomedial amygdaloid nuclei, the medial and interposed nuclei of the cerebellum, and several additional areas of the brainstem. In some of the cases, lumbar injections of wheat germ agglutinin conjugated to horseradish peroxidase were combined with a contralateral hemisection of the thoracic cord in order to determine laterality. The origins of monoaminergic projections were assessed using the retrograde transport of True Blue from the cervical cord and immunofluorescence for serotonin and tyrosine hydroxylase. Our results suggest that the origins of supraspinal projections to the spinal cord of Monodelphis are similar to those described for the North American opossum, Didelphis virginiana. Differences appear to exist, however, particularly in the amount of isocortex containing corticospinal neurons, the existence of spinal projections from the amygdala and preoptic area, the degree of rubrospinal somatotopy, and the origins of certain monoaminergic projections.
Animals↗
Development of catecholaminergic projections to the spinal cord in the North American opossum, Didelphis virginiana.
The intent of our study was to determine when catecholaminergic axons grow into each of their adult targets in the spinal cord of the North American opossum (Didelphis virginiana) and to identify the origin of catecholaminergic axons in the lumbosacral cord at different stages of development. Tyrosine hydroxylase-like immunoreactive axons, presumed to be catecholaminergic, were demonstrated at different stages of development by the indirect antibody peroxidase-antiperoxidase technique of Sternberger. The neurons giving rise to such axons in the lumbosacral cord were identified by using the retrograde transport of Fast Blue and immunofluorescence for tyrosine hydroxylase-like immunoreactive neurons. At birth, 12-13 days after conception, tyrosine hydroxylase-like immunoreactive axons are present in the marginal zone throughout the length of the spinal cord. Such axons are particularly numerous in the dorsolateral marginal zone, the region containing most of them in adult animals. By postnatal day 3, a few immunoreactive axons are present in the intermediate (mantle) zone of the spinal cord; and by postnatal day 8, they are most concentrated in the presumptive intermediolateral cell column. Laminae I and II of the dorsal horn are not innervated by such axons until approximately postnatal day 15. By postnatal day 44, the distribution of tyrosine hydroxylase-like immunoreactive axons in the spinal cord resembles that in adult animals, although some areas may be hyperinnervated. At birth, tyrosine hydroxylase-like immunoreactive cell bodies are present in all of the brainstem areas providing catecholaminergic projections to the spinal cord in adult animals (Pindzola et al.: Brain Behav. Evol. 32:281-292, '88); and by at least postnatal day 5, lumbosacral injections of Fast Blue retrogradely label tyrosine hydroxylase-like immunoreactive neurons in all such areas. Retrogradely labeled immunoreactive neurons were also found in areas that do not contain them in adult animals. Such areas include the dorsal part of the nucleus coeruleus and certain areas of the reticular formation. During development, spinally projecting tyrosine hydroxylase-like immunoreactive neurons are numerous medial to the nucleus ventralis lemnisci lateralis (the paralemniscal region), whereas only a few are present in the same location in adult animals. Our results suggest that catecholaminergic axons grow into the spinal cord prenatally, that they innervate their adult targets postnatally and over an extended time period, and that during some stages of development they originate from areas that do not supply them in the adult animal.
Aging↗
Origins and terminations of bulbospinal axons that contain serotonin and either enkephalin or substance-P in the North American opossum.
We have shown previously that some enkephalin, substance-P, and serotoninergic neurons in the medullary raphe and adjacent reticular formation project to the spinal cord in the opossum. In the present study we have combined the retrograde transport of True Blue and immunofluorescence histochemistry to determine whether methionine enkephalin or substance-P containing bulbospinal neurons are serotoninergic. Furthermore, we have used the same immunofluorescence protocol to determine whether spinal axons contain the same substances. Neurons that immunostained for both enkephalin and serotonin were observed in many brainstem nuclei. However, those that projected to the spinal cord were limited to the nuclei raphe magnus and obscurus, and the ventral part of nucleus reticularis gigantocellularis, pars ventralis. Neurons that immunostained for both substance P and serotonin were fewer in number, but some of the ones in the above nuclei and within the nucleus raphe pallidus, projected to the spinal cord. Spinal axons exhibiting both enkephalin- and serotonin-like immunoreactivity were observed in the superficial laminae of the dorsal horn, lamina X, and the intermediolateral cell column, whereas those showing both substance-P and serotonin-like immunoreactivity were seen primarily in lamina X, the intermediolateral cell column, and the ventral horn. Some of the axons in the ventral horn were in close apposition to presumed motoneurons. Comparison of the above results with those obtained from previous studies of bulbospinal projections has allowed us to infer the origins of axons that innervate different spinal targets.
Animals↗
Physiologically significant inhibitory hypothalamic action of substance P on prolactin release in the male rat.
To evaluate a possible physiological role of endogenous substance P (SP) in the control of prolactin (PRL) release, conscious adult male rats were given injections of a specific antiserum against SP (anti-SP) into the third ventricle (3 microliters) or intravenously (0.5 ml). Third-ventricular injection of anti-SP induced a significant increase in plasma PRL levels when compared to values in control animals injected with normal rabbit serum (p less than 0.02). Plasma PRL concentrations were significantly elevated within 2 h after injection of antiserum and remained elevated for the 4-hour duration of the experiment. In contrast, injections of large doses of anti-SP intravenously had no effect on plasma PRL levels. In order to confirm the effect of SP itself, synthetic SP was injected intravenously and intraventricularly. Opposite effects of SP on PRL release were observed after intravenous and intraventricular injections of low or high doses of the peptide. A lower dose of SP (10 ng, 7.42 pmol) injected into the third ventricle suppressed the release of PRL (p less than 0.01), whereas higher doses (1 microgram, 0.74 nmol, or 5 micrograms, 3.71 nmol) had a stimulatory effect on PRL release (p less than 0.01). Similarly, a low dose of SP (0.1 microgram, 0.07 nmol) injected intravenously lowered plasma PRL (p less than 0.05). Large doses of intravenous SP (50 micrograms, 37.1 nmol) dramatically stimulated PRL release (p less than 0.001). To evaluate a possible direct action of SP on PRL release from the anterior pituitary, the peptide was incubated with dispersed anterior pituitary cells for 1 h.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
The early development of major projections to the dorsal striatum in the North American opossum.
We have employed immunocytochemical and axonal transport techniques to study the development of major projections to the dorsal striatum of the North American opossum. The opossum is born in a very immature state, 12-13 days after conception, and climbs into an external pouch where it remains attached to a nipple for several months. Its immaturity at birth and its protracted postnatal development make the opossum a good model for developmental studies. Although tyrosine hydroxylase-like immunoreactive (TH-LI), presumably dopaminergic, neurons were present in the ventral mesencephalon at birth (the presumptive substantia nigra and ventral tegmental area), there was no evidence for TH-LI axons in the striatal anlage. By postnatal day (PD)6, a few immunostained axons were found within the putamen. The subsequent growth of TH-LI axons into the striatum followed general caudal to rostral and ventrolateral to dorsomedial gradients and, at any age, they were most numerous in the areas exhibiting the greatest cytodifferentiation. By estimated (E)PD45, TH-LI axons were present in most, if not all, areas of the striatum. Serotoninergic (5-HT)-LI axons were found lateral to the presumptive striatum at birth but not within it. By PD7, however, a few 5-HT-LI axons could be identified in the putamen. The growth of 5-HT-LI axons into the striatum generally followed the same gradients described for TH-LI axons although at all ages their density was much less. Using the orthograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), evidence was obtained for the existence of thalamostriatal projections by PD5 and for corticostriatal projections by PD10. Crossed corticostriatal projections were present by EPD23. Our results suggest that the development of major projections to the striatum occurs postnatally in the opossum, rather than prenatally as in placental animals. The timetable for striatal innervation is discussed in light of the developmental sequences established for other motor circuits.
Animals↗
An electron microscopic study of somatostatin immunoreactive structures in lamina II of the rat spinal cord.
We are reporting the results of a light and electron microscopic study of somatostatin (SOM) immunoreactive (I) structures in lamina II of the lumbar spinal cord of the rat. At the light microscopic level, the observed distribution and morphology of SOM-I cell bodies and fibers confirmed published studies. At the electron microscopic level, SOM immunostaining in perikarya was localized to the golgi region. Immunostaining in cell bodies could be enhanced by colchicine treatment and after such treatment, it was noticeably increased in the cytoplasm. Synaptic contacts on SOM-I cell bodies were rare and SOM-I axons contacted unlabeled somata in lamina II. Some SOM-I dendrites participate in glomerular arrangements and they exhibited postsynaptic densities adjacent to the central profile of the glomerulus. Nonglomerular SOM-I dendrites and spines were postsynaptic to vesicles containing axons. Vesicle containing SOM-I axons presynaptic to larger dendrites were also observed in the outer portion of lamina II. Somatostatin has been implicated in nociception and some of the SOM-I structures reported here may be the anatomical substrates for SOM-induced analgesia.
Animals↗