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A K Hall

Publications and source records attributed to A K Hall.

At least 37 records · Page 2Linked to original sources

SA-1 antigen expression in small intensely fluorescent cells is associated with proliferation.

Sympathetic neurons, chromaffin cells, and small, intensely fluorescent (SIF) cells are thought to derive from a common sympathoadrenal precursor cell. Sympathoadrenal precursor cells and adrenal chromaffin cells have been shown to react with an antibody, SA-1; we now report that, like sympathoadrenal precursors, SIF cells during their proliferative phase transiently possess this epitope. Precursors and SIF cells were identified in double-label studies of the superior cervical ganglion (SCG) using SA-1 and an antibody that identifies a noradrenergic trait, tyrosine hydroxylase (TH). At E16 when the earliest SIF precursors were detected and at birth, while postmitotic principal neurons had lost SA-1 reactivity, many SIF cells expressed both TH and SA-1. As development proceeded, the proportion of SIF cells expressing only TH increased. In addition, some small SIF-like cells possessed only SA-1 reactivity at birth. Some SIF cells at P7 possessed SA-1, but it was absent at P10 and in the adult. Bromodeoxyuridine (BrdU) was used to identify proliferating SIF cells, and SA-1+ expression was correlated with the period of SIF cell proliferation. At late embryogenesis, the proportion of SA-1+ SIF cells that possessed BrdU was relatively large (17% at E20), and decreased as SIF cell division ceased (6% at P1). Our results indicate that SA-1 is present on SIF cells when these cells are capable of cell division. In addition, mature SIF cells lack SA-1 and are therefore antigenically distinct from the sympathoadrenal precursor. These data suggest that the expression of SA-1 is correlated with the ability of sympathoadrenal cells to proliferate, in the SCG early during embryogenesis, chromaffin cells both during embryogenesis and in the adult, and SIF cells during their transient period of division in the SCG.

Animals↗

Retinoids and a retinoic acid receptor differentially modulate thymosin beta 10 gene expression in transfected neuroblastoma cells.

1. Investigations have demonstrated that the gene encoding thymosin beta 10 (a 43-amino acid member of a family of related proteins originally described in the rat immune system) is a target for morphogenic retinoids in both human and rat neuroblastoma cells. 2. Structure-activity studies revealed that the stimulatory actions of retinoids upon the thymosin beta 10 gene reflect the differing affinities of retinoid analogues for a retinoic acid receptor. 3. To examine further the possibility that the trophic actions of retinoic acid upon expression of the thymosin beta 10 gene involved retinoid receptors, neuroblastoma cells were transiently transfected with an expression vector encoding the nuclear retinoic acid receptor (alpha) protein. 4. Northern blot and slot-blot analyses revealed that neuronal cells overexpressing RAR alpha-mRNA exhibited an enhanced sensitivity to exogenous and endogenous retinoic acid in terms of thymosin beta 10 mRNA. Although the RAR-alpha gene was expressed (at low levels) a priori in these neuroblastoma cells, retinoic acid (2 x 10(-7) M for 3 days) slightly stimulated RAR-alpha-mRNA accumulation. 5. Collectively, these findings indicate the retinoic acid receptor (alpha) is regulated by retinoid acid and that the developmentally regulated, retinoid-responsive thymosin beta 10 gene is a target for this nuclear transcription factor in cells derived from the neural crest.

Animals↗

Division and migration of satellite glia in the embryonic rat superior cervical ganglion.

While distinct precursors committed to a neuronal or glial cell fate are generated from neural crest cells early in peripheral gangliogenesis, little is known about the subsequent generation and maturation of young satellite glia from restricted glial precursor cells. To examine the division and migration of glial precursor cells and their satellite cell progeny, morphological, immunocytochemical and culture techniques were applied to the developing rat superior cervical ganglion. At embryonic day (E)18.5, numerous clusters of nonneuronal cells appeared transiently in the ganglion. Individual cells with a similar morphology were present in E16.5 ganglia, and are likely to represent the precursor cells which generate these clusters. The clustered cells were distinguishable from neighbouring neurons as well as from endothelial cells and fibroblasts. Morphologically similar cells were present in nerve bundles at E18.5 and surrounding principal neurons and nerve bundles in the adult ganglion. Double-label studies of the E18.5 ganglion with tyrosine hydroxylase to identify noradrenergic neurons and propidium iodide counterstaining to visualize all cell nuclei revealed that the cells in clusters stained with propidium iodide but lacked tyrosine hydroxylase immunoreactivity. To determine if cell clusters arose from division, bromodeoxy-uridine, a thymidine analogue, was administered to pregnant mothers between E16.5-E18.5, and ganglionic cells examined at E18.5 both in vivo and in vitro. Numerous non-neuronal cells divided during this period in situ and composed portions of clusters. When dissociated, superior cervical ganglion satellite glia reacted with an NGF-receptor antibody (MAb 217c) and possessed a flattened shape, in contrast to bipolar Schwann cells. Over half of the 217c-immunoreactive glia at E18.5 had incorporated bromodeoxyuridine during E16.5-18.5 in vivo. At birth, non-neuronal cells were no longer grouped in clusters, but were associated with neuronal cell bodies and processes. These findings suggest that, between E16.5-E18.5, glial precursors divide rapidly to form clusters, and that, after the peak of neurogenesis, daughter cells migrate within the ganglion to associate with nerve cell bodies and processes where proliferation continues at a slower rate. Distinct cellular and molecular interactions are likely to trigger the initial rapid division of glial precursors, initiate their migration and association with neuron cell bodies, and control their subsequent slower division.

Animals↗

Influence of cyclic AMP and serum factors upon expression of a retinoid-responsive gene in neuroblastoma cells.

Cyclic AMP can profoundly influence the growth and differentiation of neuronal cells in culture. In this study, the relationship between this second messenger signal transduction pathway, cell differentiation, and the expression of a retinoid-responsive, thymosin beta-10 gene was examined. Thymosin beta-10 and cognate mRNA were expressed at high levels in actively proliferating rat B104 neuroblastoma cells cultured in medium containing 10% FCS. These cells were induced to differentiate in the presence of the cAMP analog N6, 2'-O-dibutyryladenosine 3':5'-cyclic monophosphate (Bt2-cAMP) (1 mM) and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) (100 microM). Expression of thymosin beta-10 mRNA was markedly inhibited (greater than 90% and 70%, respectively) by these compounds. Addition of sodium butyrate (NaB, 1 mM) indicated that at least part of the inhibitory actions of Bt2-cAMP were due to esterase-induced release of butyrate from this compound. Adenosine (50 microM), a metabolic precursor to endogenous cyclic AMP, also inhibited accumulation of thymosin beta-10 mRNA (to less than 70% of control levels). The inhibitory action of Bt2-cAMP upon thymosin beta-10 mRNA levels was time dependent; levels were inhibited by greater than 50% 24 hours after addition of the cAMP analog and by greater than 90% after 72 hours. Serum starvation (0.2% FCS for seven days) provoked a marked increase in neurite out-growth; this morphological change was also accompanied by a modest inhibition of thymosin beta-10 mRNA accumulation. These findings together with previous observations imply that both cyclic AMP-dependent and retinoid-responsive mechanisms coordinate thymosin beta-10 gene expression during neuroembryogenesis.

1-Methyl-3-isobutylxanthine↗

Expression of thymosin beta-4 and related genes in developing human brain.

The retinoic acid-responsive thymosin beta-10 gene is known to be developmentally regulated in the human brain. We now report the novel finding that thymosin beta-4, a structurally related 5-kDa actin-sequestering protein, is also subject to a similar but not identical pattern of expression during normal human neuroembryogenesis. However, while thymosin beta-10 mRNA was undetectable (by northern blot analysis) in adult human brain, levels of thymosin beta-4 mRNA, although greatly reduced, were still present. Moreover, a novel thymosin beta-10-like gene was also found to exhibit a unique stage-specific expression during early human neural development. These experiments, together with previous findings, indicate that the products of the two thymosin genes, possibly in association with cytoskeletal elements, may play different roles during early neuroembryogenesis and neural maturation.

Adult↗

Bulimia nervosa. Four uncommon subtypes.

The histories and psychological profiles of more than 500 patients meeting DSM-III-R criteria for bulimia nervosa were reviewed. A total of 310 patients demonstrated the most characteristic pattern of bulimia, with finger-induced purging and occasional diet pill, diuretic, or laxative abuse. Seventeen patients reported binge eating with no self-induced vomiting but with severe laxative abuse (i.e., greater than or equal to 50 laxatives daily). A total of 126 patients reported bulimia with finger-induced purging and regular mild (i.e., 2-3 daily) laxative abuse. Eight patients reported bulimia without finger-induced purging, diuretic, or laxative abuse but with the regular abuse of ipecac as a means of inducing vomiting. Four clinical subtypes of bulimia were seen. These were overt bulimia, which occurred in 8.9% of the sample; obsessive-ritualistic bulimia, which occurred in 2% of the sample; sexually evocative bulimia (Fatal Attraction Syndrome), which occurred in 2.9% of the sample; and masochistic bulimia, which occurred in 4.9% of the sample. Each of these subtypes of bulimia are described and defined. The characteristic psychologic profile, clinical features, and implications for treatment and research are discussed.

Acting Out↗

Differential expression of thymosin genes in human tumors and in the developing human kidney.

Thymosins beta 4 and beta 10 are 2 structurally related polypeptides originally defined in the rat immune system. To date, no truly unambiguous functions have been formally ascribed to these small (less than 4.9 kDa) acidic proteins. Previous research has demonstrated relationships between expression of these genes and cell growth/differentiation. These observations prompted the present study which has used cDNA and synthetic oligonucleotide probes in combination with high-performance liquid chromatography (HPLC) to examine the differential expression of these 2 genes in normal and neoplastic human tissues and in the developing human kidney. Low levels of beta 4 and beta 10 mRNA species prevailed in normal tissues; in contrast, these gene transcripts were notably more abundant in malignant renal tumors and in the normal human embryonic kidney. These findings show that the thymosin beta 4 and beta 10 genes are constitutively expressed at higher levels in embryonic/neoplastic as compared to normal/benign tissues and that thymosin in beta 10 in particular may be a new molecular marker for renal-cell carcinoma as well as other malignancies.

Aging↗

Acute actions of 1,25-dihydroxyvitamin D3 upon chick pancreatic calbindin-D28K.

We have compared the relative responsiveness of pancreatic, intestinal and renal tissue calbindin-D28K protein content to the stimulatory actions of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] in vitamin D-deficient (-D) chicks. Tissue concentrations of calbindin-D28K were undetectable in the -D chick intestine but present, albeit at low concentrations (less than 1 microgram CaBP/mg protein) in the -D kidney and pancreas. Intestinal, pancreatic and renal calbindin-D28K content was stimulated 318, 9.8 and 2.9 fold respectively, 48 hours after -D chicks received a single dose of 1,25(OH)2D3 [6.5 nmol/animal]. The pancreatic calbindin-D28K content could be significantly stimulated as early as 5 hours after 1,25(OH)2D3 administrations in vivo. These findings support the contention that the pancreas is a target for vitamin D, and is consistent with the view that calbindin-D28K plays a role in normal pancreatic functions.

Animals↗

Vitamin D-independent expression of chick brain calbindin-D28K.

A combination of calbindin-D28K-specific cDNA probes and polyclonal antisera were used to investigate expression of the calbindin-D28K in the vitamin D-deficient avian brain in vivo in response to pharmacological doses of the vitamin D3 metabolite, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Serum calcium levels were stimulated (2-fold) and intestinal calbindin-D28K expression (between 10- and 30-fold) by 1,25(OH)2D3 (6.5 nmol/animal) after 12 h. In marked contrast, steady-state whole brain levels of calbindin-D28K as judged by enzyme-linked immunoassay (ELISA) remained constant. Northern gel analysis revealed that three species of calbindin-D28K mRNA (2.0, 2.6 and 3.1 kb) were present a priori in the vitamin D-deficient chick brain and that administration of pharmacological doses (6.5 nmol/animal) of 1,25(OH)2D3 failed to influence their relative abundance. Separate but parallel dot blot hybridization analyses also confirmed that brain calbindin-D28K-mRNA levels were not influenced by 1,25(OH)2D3. These experiments demonstrate at the molecular level that, in contrast to the intestine, the gene encoding calbindin-D28K in the brain is regulated by mechanism(s) or factors which are independent of vitamin D status.

Animals↗

Developmental regulation of thymosin beta 10 mRNA in the human brain.

A specific thymosin beta 10 cDNA restriction fragment was used to monitor thymosin beta 10 mRNA levels in the brain during normal human neuroembryogenesis. Northern analysis revealed a single hybridizing 600 nucleotide mRNA species which was abundant in embryonic/fetal human brain, yet absent in the adult tissue. These findings suggest that thymosin beta 10 must play an important role in early neuroembryogenesis.

Brain↗

Differential modulation of thymosin genes in the immature rat ovary by gonadotropins.

A cloned thymosin beta-10 cDNA and a synthetic oligonucleotide specific for the thymosin beta-4 gene were used to study the in vivo expression of these two genes in the immature rat ovary in response to exogenously administered gonadotropins. Despite the fact that both genes were co-expressed in the rat ovary, it became evident that they exhibit distinctly unique differential responses to in vivo hormonal challenge. Administration of pregnant mare's serum gonadotropin (PMSG) to immature rats provoked a pronounced stimulation of ovarian thymosin beta-10 expression, the maximal effect (2- to 4-fold) of which coincided with the time at which folliculogenesis was also maximally enhanced. In contrast, the transcriptional status of the thymosin beta-4 gene varied little in response to the PMSG. Administration of human chorionic gonadotropin (hCG) to PMSG-primed rats inhibited ovarian thymosin beta-10 but stimulated thymosin beta-4 gene expression. These findings suggest that despite a mutually high degree of homology, these two proteins may, under the influence of gonadotropins, play independent roles in normal ovarian function.

Animals↗

Early commitment of precursor cells from the rat superior cervical ganglion to neuronal or nonneuronal fates.

To determine whether postmigratory neural crest cells retain the capacity to give rise to multiple cell types, the clonal progeny of embryonic rat superior cervical ganglion (SCG) cells were examined in culture. Double labeling with BrdU and neurofilament antibodies demonstrated that neuron precursors from the E14.5 SCG continued to proliferate for several days in culture. Using the BAG retrovirus to examine the progeny of single cells, we obtained several kinds of distinct clones from SCG cultures after 3 days. At E14.5, during peak neurogenesis in vivo, neuron-containing clones composed of one to seven cells were common. At E17.5, after neurons have been born in vivo, most clones in vitro contained flat cells, primarily reflecting glial cell division. Even in cultures from E13.5 ganglia, mixed clones containing neurons and flat cells were rarely observed. These observations suggest that neuronal and nonneuronal cell precursors are specified during or before early gangliogenesis.

Animals↗

Developmental regulation of beta-thymosins in the rat central nervous system.

HPLC analysis of guanidinium hydrochloride extracts of neonatal and adult rat brain revealed a polypeptide that is present in high concentration in the immature nervous system, but whose levels decline dramatically in the adult. This polypeptide has been isolated and its complete amino acid sequence determined by gas-phase Edman degradation following specific chemical and enzymatic cleavages. The molecule is identified as thymosin beta 10, a member of a multigene family that encodes a structurally conserved series of small acidic polypeptides of uncertain function. Thymosin beta 10 is present in the developing nervous system as early as embryonic day 9. Levels subsequently increase to peak values between embryonic day 15 and postpartum day 3, before falling to adult values (about a 20-fold reduction) by postpartum day 14. The elevated levels of thymosin beta 10 in fetal and neonatal brain correlate with high levels of thymosin beta 10 mRNA, whereas the low values of the polypeptide in the adult and juvenile are mirrored by an approximate 15-fold reduction in specific mRNA. In comparison, the levels of thymosin beta 4 polypeptide, a homologue of thymosin beta 10, only decline by about 20% during the same developmental period. However, the mRNA encoding thymosin beta 4 is elevated in fetal brain, and its levels decrease approximately four-fold to a stable value around the time of birth. The reason for this discrepancy between thymosin beta 4 protein and mRNA levels is unknown. Thymosin beta 10 can also be detected by HPLC in fetal liver, where levels are approximately 5% of those in brain. In liver, thymosin beta 10 also declines following birth. It is concluded that beta-thymosin expression (as measured by steady-state mRNA and polypeptide levels) is both up- and down-regulated during different phases of maturation of the mammalian nervous system.

Aging↗

Retinoic acid regulates thymosin beta 10 levels in rat neuroblastoma cells.

A small acidic polypeptide, termed thymosin beta 10, has been identified and is present in the nervous system of the rat by the ninth day of gestation. Thymosin beta 10 levels rise during the remaining days of life in utero, and then decline to nearly undetectable values between the second and fourth week post partum. The present study investigates the possible developmental signals and mechanisms that might regulate the expression of thymosin beta 10 during neuroembryogenesis. Many cell lines derived from tumors of the central nervous system express thymosin beta 10, as well as its homologue gene product, thymosin beta 4. Because some of these cell lines respond to exogenously applied agents by increasing their apparent state of differentiation, we have determined whether thymosin beta 10 levels are coordinately modulated. In several neuroblastomas, including the B103 and B104 lines, retinoic acid elicits a time- and dose-dependent increase in the content of thymosin beta 10, but not that of thymosin beta 4. The increase in thymosin beta 10 polypeptide is associated with a marked increase in the specific mRNA encoding this molecule. The mRNA for thymosin beta 4 is unaffected by retinoic acid. This is in contrast with the situation in vivo, where the expression of both genes decreases after birth. Other agents that influence the morphology of B104 cells, such as phorbol esters and dibutyryl cyclic AMP, have no influence on beta-thymosin levels. A range of steroids, which like retinoids act upon nuclear receptors, was also inactive. The stimulatory action of retinoic acid is detectable within 4 h, and thymosin beta 10 peptide levels continue to rise for at least 4 days. The influence of the isoprenoid is fully reversible and exhibits structural specificity. We believe that this culture system is mimicking the early rising phase of thymosin beta 10 levels in brain and that endogenous retinoids may be candidate physiological regulators of this gene.

Animals↗

Thymosin gene expression is modulated by pregnant mare's serum gonadotropin, human chorionic gonadotropin, and prostaglandin F2 alpha in the immature rat ovary.

We have investigated thymosin beta 10 mRNA levels in the PMSG/hCG-treated immature rat ovary. Thymosin beta 10 mRNA was constitutively expressed as a single (greater than 600-nucleotide) abundant transcript in the immature rat ovary. Administration of a single dose (50 IU/rat) of PMSG to immature rats resulted in a gradual increase in steady state ovarian thymosin beta 10 mRNA content detectable as early as 12 h and maximal (2-to 3-fold stimulation above preinjection levels) 48 h after PMSG treatment. Ovarian thymosin beta 10 mRNA levels declined thereafter. In separate experiments treatment of PMSG (50 IU)-primed rats with hCG (25 IU) precipitated a dramatic (80%) inhibition of ambient ovarian thymosin beta 10 protein and thymosin beta 10 mRNA; both parameters remained suppressed for the duration of the hormone-induced pseudopregnancy (15 days). HPLC analysis also indicated the presence in the ovary of thymosin beta 4, a variant member of the same protein family; in general, ovarian thymosin beta 4 levels fluctuated in a manner reciprocal to that exhibited by thymosin beta 10. A luteolytic dose of prostaglandin F2 alpha (500 micrograms/rat) had little impact on ovarian thymosin beta 10 gene expression. These findings show that 1) PMSG stimulation of ovarian thymosin beta 10 biosynthesis involves increased expression of the thymosin beta 10 gene; 2) decreased expression of thymosin beta 10 is associated with luteinization, while increased thymosin beta 4 levels characterize this process; and that 3) thymosin beta 4 and beta 10 are coexpressed in this tissue and may play a significant but, as of yet, undefined role(s) in the ovary.

Animals↗

Principal neurons and small intensely fluorescent (SIF) cells in the rat superior cervical ganglion have distinct developmental histories.

Sympathetic ganglia contain 2 adrenergic derivatives of the neural crest: principal neurons and small, intensely fluorescent (SIF) cells. The developmental mechanisms responsible for the generation of these 2 cell classes in vivo are not well understood. To examine the possible developmental and lineage relationships between differentiating principal neurons and SIF cells, a fluorescence microscopic study utilizing antibodies against tyrosine hydroxylase (TH) and catecholamine histofluorescence has been combined with the ultrastructural examination of embryonic and postnatal rat superior cervical ganglia (SCG). On embryonic day 12.5, before neuroblasts had become postmitotic, the cells in the SCG possessed intense TH immunoreactivity and had weak to bright catecholamine histofluorescence, but no cells displayed the fine structure of mature SIF cells or neurons. At embryonic days 16.5 and 18.5, postmitotic principal neurons expressed more moderate levels of TH and catecholamines characteristic of the late embryonic and postnatal SCG. By contrast, a small number of cells containing intense TH or catecholamine fluorescence were present in embryonic day 16.5 and older ganglia. Almost all of the intensely fluorescent cells observed were found apposed to capillaries within the ganglion. These embryonic intensely fluorescent cells were larger than SIF cells seen postnatally. Ultrastructural examination of developing ganglia confirmed that cells containing numerous large, dense-cored vesicles (LDCVs) were a prominent feature of ganglia that also contained intensely fluorescent cells. In addition, some embryonic cells containing LDCVs were mitotic. From these and other studies, it seems likely that during development, neuron precursors, in response to differentiation factors such as fibroblast growth factor (FGF) and/or NGF, acquire overt neuronal traits and become postmitotic. Subsequently, cells resembling mature SIF cells appear next to blood vessels, where they may have received other instructional signals such as glucocorticoids. This developmental scheme suggests that the differentiation of principal neurons and SIF cells is independently regulated, and that the ability of SIF cells to convert into principal neurons observed in vitro cannot account for the generation of neurons in vivo.

Animals↗

Retinoic acid and serum modulation of thymosin beta-10 gene expression in rat neuroblastoma cells.

A cloned thymosin beta-10 cDNA was used to study modulation of thymosin beta-10 mRNA levels in the rat B104 neuroblastoma cell line in response to retinoic acid. Northern blot analysis revealed the presence of a single greater than 600-nucleotide thymosin beta-10 mRNA species that was constitutively expressed in proliferating neuroblastoma cells. Addition of retinoic acid to the culture medium induced a dose- and time-dependent increase in thymosin beta-10 mRNA abundance. Additional studies showed that although thymosin beta-4 and beta-10 are coexpressed in this cell line, the stimulatory action of retinoic acid is specific for the thymosin beta-10 gene. Serum was found to augment the stimulatory action of retinoic acid. Blockade of protein synthesis with cycloheximide abrogated the stimulatory action of retinoic acid upon thymosin beta-10 mRNA accumulation; this observation suggests that activation of the thymosin beta-10 gene in this cell line by retinoic acid is dependent upon the de novo synthesis of a labile protein. Collectively, these findings demonstrate that the developmentally regulated thymosin beta-10 gene is a target for morphogenic retinoids in cells derived from the neural crest.

Animals↗