Search PubMed⌕ Search

Biomedical subjects

P H Patterson

Publications and source records attributed to P H Patterson.

At least 55 records · Page 3Linked to original sources
PubMed1997

An antibody to the tetraspan membrane protein CD9 promotes neurite formation in a partially alpha3beta1 integrin-dependent manner.

The tetraspan cell surface glycoprotein, CD9, has been implicated in cellular signaling during growth and differentiation in the hematopoietic and nervous systems. Because CD9 expression is induced early in development in sensory and sympathetic neuroblasts, we investigated the role of CD9 in neurite outgrowth. We plated dissociated cells from neonatal sympathetic ganglia on immobilized anti-CD9 antibodies or antibodies against other cell surface molecules. We show here that B2C11, an anti-CD9 antibody that has been shown previously to activate Schwann cells in vitro, promotes robust neurite outgrowth from sympathetic neurons that is greater than that on other antibody surfaces and is comparable to neurite outgrowth on a collagen substratum. In addition, B2C11 causes dramatic morphological changes in neurons and glia from dissociated ganglia, including a flattening of these cells. Because CD9 interacts with integrins in many cell types including Schwann cells, and specifically with the alpha3beta1 integrin in some cells, we tested whether the effect of B2C11 on neurite outgrowth is mediated by this integrin. An anti-alpha3beta1 antibody, Ralph 3-1, attenuates the extent of neurite outgrowth on B2C11 and collagen, but not on laminin. Because the alpha3beta1 integrin has been shown to mediate neurite outgrowth on different substrates, these results provide a functional significance for the CD9-alpha3beta1 interaction; downstream signaling may be activated by this cis interaction on the cell surface in response to external cues that promote neurite outgrowth.

Animals↗
PubMed1997

Leukemia inhibitory factor is expressed in astrocytes following cortical brain injury.

The neuropoietic cytokine leukemia inhibitory factor (LIF) can act as a trophic factor, enhancing neuronal survival, and as a differentiation factor altering neuronal and glial gene expression. LIF also plays a role in the response to injury of the peripheral nervous system, as indicated by an increase in the amount of its mRNA within nonneuronal injury response in LIF knock-out mice. To determine if LIF is regulated after injury to the central nervous system, we surgically lesioned the cortex in adult rat brain. Using a quantitative RNAse protection assay, we find that LIF mRNA increases 30-fold following injury. The amount of this transcript goes up within 6 h after injury, reaches a peak at 24 h and returns to baseline by 7 days postlesion. In situ hybridization analysis reveals LIF transcript-containing cells scattered throughout the ipsilateral cortex close, but not immediately adjacent to the lesion site. Double-labeling with a variety of antibodies reveals that LIF mRNA is induced in GFAP-positive astrocytes as well as in a small number of microglial cells. The striking induction of LIF transcripts in glia suggests that this cytokine may play a key injury-response role in the CNS as it does in the PNS, where LIF has been demonstrated to regulate neuropeptide expression both in vivo and in vitro.

Animals↗
PubMed1997

LIF is an autocrine factor for sympathetic neurons.

Leukemia inhibitory factor (LIF) alters neuronal phenotypes both in vitro and in vivo. Since it can be produced by glia and other nonneural cells, LIF is a candidate target-derived differentiation factor as well as an injury-response factor. We here provide evidence that LIF can be produced by neurons and can act on the neurons that produce it. A reverse transcriptase-polymerase chain reaction assay detects LIF mRNA in rat sympathetic neuron cultures, and in situ hybridization combined with MAP2 immunocytochemistry indicates that most of the cells expressing LIF mRNA are, in fact, neurons. The neuronal lysate as well as the conditioned medium contains proteins that are specifically recognized by anti-LIF antibodies, and these antibodies also specifically stain the cultured neurons. In addition, concentrated sympathetic neuron conditioned medium can mimic the effects of LIF, and incubation of high-density sympathetic neuron cultures with anti-LIF antibodies reduces basal expression levels of LIF target genes such as particular neuropeptides, indicating that the endogenously produced cytokine is acting on the neurons under these conditions. Since we show that LIF transcript is expressed in sympathetic and sensory neurons in vivo as well, LIF could act in an autocrine fashion under a variety of physiological conditions.

Animals↗
PubMed1997

Cardiotrophin-1 induces the same neuropeptides in sympathetic neurons as do neuropoietic cytokines.

Cardiotrophin-1 (CT-1) was cloned from mouse embryoid body for its ability to induce growth of heart cells. Predictions of its secondary structure indicate that CT-1 belongs to a family of cytokines with a four-helical bundle structure, and sequence comparisons reveal a weak homology to leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF). Using a reverse transcriptase-polymerase chain reaction assay with rat sympathetic neuron cultures, we find that CT-1 induces and suppresses the expression of the same set of neuropeptide and neurotransmitter synthetic enzyme mRNAs as do LIF and CNTF. In addition, the effects of CT-1 and LIF are not additive, and CT-1 does not require a GPI-linked component to mediate its actions. Our functional data confirm that CT-1 is a member of the neuropoietic cytokine family and suggest that the CT-1 receptor complex contains the gp130 signal transducing component.

Animals↗
PubMed1997

Preservation of hatchery waste by lactic acid fermentation. 1. Laboratory scale fermentation.

In Experiment 1, two types of hatchery wastes, including cockerel chicks (CC), and shell waste (SW) blended with CC (60:40 CC:SW), were ground and mixed with a by-product carbohydrate (5, 10, and 15% wt/wt) with or without a bacterial culture and fermented for a period of 21 d. Cockerel chicks fermented with 15% carbohydrate and the culture had a pleasant smell and little H2S production. Elevated H2S concentrations were recorded for CC:SW samples at all carbohydrate levels when the culture was not added. No NH3 was detected from any treatments during fermentation. The addition of culture to the CC and CC:SW by-products resulted in pH values lower (P < 0.05) than those without culture on Day 21, and the 15% carbohydrate treatment significantly reduced pH beyond the 5% carbohydrate. Final proximate composition of CC and CC:SW samples with culture were not significantly different from those without culture added. In Experiment 2, carbohydrate was added at 10.0, 13.3, 16.7, and 20.0% wt/wt to CC and CC:SW in the presence of the bacterial culture. Shell waste alone was fermented with 15, 20, and 25% carbohydrate and the culture. Moisture level in this experiment was adjusted to approximately 70% for all treatments. The lowest pH for the CC and CC:SW treatments was observed at the 16.7% carbohydrate level. Shell waste pH was better maintained at the 20 and 25% carbohydrate levels. After fermentation for 21 d CC, CC:SW and SW treatments from Experiments 1 and 2 contained negligible Escherichia coli, and no Salmonella were detected.

Analysis of Variance↗
PubMed1997

Preservation of hatchery waste by lactic acid fermentation. 2. Large-scale fermentation and feeding trial to evaluate feeding value.

Two waste streams from a Leghorn hatchery were preserved and recycled by fermentation with a by-product carbohydrate and extrusion processing into new feed ingredients that were evaluated with broiler chickens. Cockerel chicks (CC) and a 60:40 ratio of CC:shell waste (CC:SW) were fermented in 189-L barrels for 21 d following grinding, then mixing with a liquid culture (0.2%) and carbohydrate source at 15 and 16.66%, respectively. At 2 wk, pH was 4.44 and 5.09 for the CC and CC:SW products compared with higher values of 6.54 and 6.98 for the raw ingredients at the onset. Negligible hydrogen sulfide and no ammonia gas were recorded during the fermentation period. At 21 d, the fermented CC and CC:SW were extruded, dried, and ground to meals containing CP and TMEn levels of 47.4%, 3,187 kcal/kg, and 33.1%, 2,696 kcal/kg, respectively. Broiler chickens were fed a control diet and the CC (5 and 10%) and CC:SW (2.5 and 5%) ingredient diets with corn and soybean meal for 6 wk to evaluate feeding value and carcass yield. Body weight, gain and feed conversion at 42 d for birds fed diets supplemented with CC or CC:SW at all levels were comparable to those of the control. Diets supplemented with hatchery by-product had no negative effect on carcass measurements except ready to cook carcass and wing yield, which were significantly greater for the 10% CC:SW birds than for the control. These data indicate that nutrient dense hatchery by-products can be preserved with fermentation up to 21 d and support broiler live performance and carcass yield as dietary ingredients equal to or better than a corn-soybean meal control.

Analysis of Variance↗
PubMed1996

Neuronal expression of leukemia inhibitory factor (LIF) in the rat brain.

Very little is known about the action of the leukemia inhibitory factor (LIF) in the central nervous system. To analyze LIF expression in adult rat brain, we used non-radioactive in situ hybridization with digoxigenin-labeled RNA probes, in combination with immunohistochemistry to identify the cell types expressing LIF mRNA. Hippocampal pyramidal cells, hilar interneurons and granule cells of the dentate gyrus display strong hybridization signals, while neurons of the cerebral cortex, cerebellum, cholinergic basal forebrain nuclei, and the striatum are labeled as well. The most prominent expression in cerebral cortex is in layers IV and VI. Glial fibrillary acidic protein-positive astrocytes did not display a detectable LIF hybridization signal suggesting that LIF mRNA is predominantly expressed in neurons in the normal, adult rat brain, although a presence in resting microglial cells cannot be excluded.

Animals↗
PubMed1996

Association of the tetraspan protein CD9 with integrins on the surface of S-16 Schwann cells.

The cell surface glycoprotein CD9 is a member of a group of proteins known as the "tetraspan" or "transmembrane 4 superfamily." Previous work with non-neural cells has shown that CD9 associates in cis with integrins and small GTP-binding proteins on the cell surface. To extend our recent findings showing that perturbation of CD9 alters Schwann cell adhesion, proliferation, and migration, as well as neurite outgrowth in sympathetic neurons, we have searched for CD9-associated proteins in S-16 Schwann cells. We demonstrate here that CD9 is specifically coprecipitated from S-16 cell extracts by antibodies against integrins alpha 3, alpha 6, and beta 1. In addition, double immunofluorescence labeling and co-capping experiments indicate that CD9 is specifically co-localized with these integrins on the cell membrane of S-16 Schwann cells.

Animals↗
PubMed1995

Differential regulation of motor neuron survival and choline acetyltransferase expression following axotomy.

Although it is well known that motor neuron survival following axotomy is enhanced with maturation, the ability of surviving neurons to express the cholinergic enzyme choline acetyltransferase (ChAT) following axotomy has not ben closely examined. Moreover, the utility of the facial nucleus in studies of motoneuron response to injury and to trophic factors, coupled with the increasing importance of the mouse in gene targeting, compelled us to investigate the age dependence of neuronal survival and ChAT expression in the mouse facial nucleus following axotomy. We cut the facial nerve at postnatal day (P) 4, 7, 14, 21, and 28 or in the adult and used Nissl staining and ChAT immunocytochemistry to quantitate survival and ChAT expression, respectively, following 1, 2, or 3 weeks' survival at each age. We confirm in this model that the rate and extent of motor neuron death following axotomy is reduced with increasing maturity. The surviving neurons maintain a high ChAT content through P21; however, axotomy from P28 through adulthood results in a striking reduction in ChAT immunoreactivity. That is, although axotomy at P21 results in 61% motor neuron survival, with virtually all of the surviving neurons being ChAT positive, axotomy in the adult results in 72% survival but only 9% of the neurons are ChAT positive. Thus, surviving motor neurons in the adult animals are only weakly cholinergic. These results indicate that a change in the regulation of ChAT expression occurs following P21 so that cell survival and enzyme levels are uncoupled. We suggest that the putative factor or factors that enhances motor neuron survival in maturity is not capable of maintaining ChAT expression.

Aging↗
PubMed1995

Adrenal chromaffin cells transdifferentiate in response to basic fibroblast growth factor and show directed outgrowth to a nerve growth factor source in vivo.

Chromaffin cells exposed to basic fibroblast growth factor (bFGF) in vitro express characteristics of sympathetic neurons, extend neurites, and become dependent on nerve growth factor (NGF) for survival. We explored whether the growth factor responsiveness of chromaffin cells could be exploited to enhance the transdifferentiation, neurite outgrowth and functional efficacy of chromaffin cells implanted into rats with unilateral 6-hydroxydopamine lesions. Cografts of neonatal chromaffin cells and fibroblasts genetically modified to produce bFGF were placed into the dopamine-depleted striatum of adult rats. Either control-transfected or NGF-producing fibroblasts were then transplanted 1 mm distal to the cograft. Chromaffin cells transdifferentiated under the influence of bFGF, as indicated by the growth of neurites and the expression of neuron-specific proteins. Distal grafts of NGF-producing fibroblasts successfully induced chromaffin neurites to traverse through the host parenchyma to the NGF source. In the absence of NGF fibroblast grafts, neither neurite extension nor good, long-term survival of the chromaffin-derived neurons was observed. Assessments of apomorphine-induced rotational behavior 2- and 4-weeks postgrafting revealed no behavior improvements in any of the groups. These results indicate that localized sources of growth factors are effective in inducing the transdifferentiation of grafted chromaffin cells as well as the extension of chromaffin-derived neurites into the host parenchyma. Such chromaffin cell-derived neurons are, however, functionally ineffective in this rat model of Parkinson's disease. Whether the lack of behavioral improvement reflected the tropic growth of neurites to an inappropriate striatal region or the noradrenergic nature of the chromaffin cell-derived neurons remains to be clarified. Nonetheless, these results caution that promoting transdifferentiation and neurite extension from engrafted chromaffin cells may not be sufficient to achieve desired functional effects of such grafts.

Adrenal Medulla↗
PubMed1995

FORSE-1, an antibody that labels regionally restricted subpopulations of progenitor cells in the embryonic central nervous system, recognizes the Le(x) carbohydrate on a proteoglycan and two glycolipid antigens.

A key problem in nervous system development is how distinct subpopulations of progenitor cells give rise to different adult brain structures. The labeling pattern of the FORSE-1 antibody subdivides the neuroepithelium of the embryonic forebrain into domains resembling those of certain transcription factors, suggesting that the FORSE-1 epitope may be involved in the specification of development compartments. Therefore, it is important to determine the identity of the antigen(s) recognized by FORSE-1. On immunoblots, FORSE-1 recognizes a single, high-molecular-weight species, which we have identified as phosphacan, a brain-specific chondroitin sulfate proteoglycan that binds neural cell adhesion molecules. This identification is based on cross-immunoprecipitations and immunoblotting using an anti-phosphacan antibody and FORSE-1. FORSE-1 also recognizes two major neutral glycolipids in embryonic brain. The FORSE-1 epitope is sensitive to endo-beta-galactosidase, suggesting that the epitope corresponds to a carbohydrate moiety. Moreover, immunoprecipitates of the proteoglycan bearing the FORSE-1 epitope bind antibodies that recognize the Le* carbohydrate, and immunostaining patterns of embryonic brain sections by FORSE-1 and a known anti-Le* antibody are identical. Finally, purified FORSE-1 specifically recognizes Le*-containing glycoconjugates in ELISAs. The pattern of FORSE-1 labeling, the identification of its epitope as Le*, which has implicated in cell adhesion, and the presence of Le* on phosphacan suggest that this carbohydrate epitope may play a role in adhesive interactions important for proliferation, cell migration, or axon guidance.

Animals↗
PubMed1995

Schwann cell CD9 expression is regulated by axons.

The cell-surface glycoprotein, CD9, has been implicated in hematopoietic and Schwann cell signaling in vitro. In vivo, Schwann cell CD9 expression follows a developmental time course that parallels that of myelin genes. Here we report that Schwann cell CD9 mRNA expression is regulated by axonal contact in culture and in vivo. Following adult rat sciatic nerve injury, CD9 mRNA expression in distal nerve sections is correlated with the presence of axons; CD9 is down-regulated when axons are degenerating and is reexpressed when axons are regenerating, but not if they are prevented from doing so by transection. CD9 regulation in cultured Schwann cells is also dependent on the presence of neurons such that dissociated Schwann cells down-regulate CD9 mRNA expression in culture, but in the presence of sensory neurons they continue to express CD9. Therefore, regulation of Schwann cell CD9 expression parallels that of myelin genes protein 0 and myelin basic protein in culture and in vivo. A signaling role for CD9 in nerve development and regeneration is proposed.

Animals↗
PubMed1995

Cytokines in Alzheimer's disease and multiple sclerosis.

Cytokines are well known as mediators of inflammation, and recent work has highlighted the role of these agents and inflammatory events in Alzheimer's disease and multiple sclerosis. The discovery of subclasses of T-helper cells has provided a critical framework to aid in understanding how the cytokine network regulates these diseases.

Alzheimer Disease↗
PubMed1995

CD9 plays a role in Schwann cell migration in vitro.

To identify molecules that regulate Schwann cell migration, we have generated a panel of monoclonal antibodies against Schwann cell surface antigens that modulate Schwann cell migration in in vitro bioassays. One of these antibodies, SMRA1, recognizes a 26 kDa Schwann cell surface membrane protein identified here as CD9. SMRA1 enhances Schwann cell migration on two biologically relevant substrates: living axons of cultured dorsal root ganglion neurons, and cryostat sections of sciatic nerve. This CD9-induced regulation of Schwann cell motility is correlated with a rise in cytosolic calcium and enhanced tyrosine phosphorylation of several Schwann cell proteins. These results, together with the findings of Hadjiargyrou and Patterson (1994), implicate CD9 as an important regulator of Schwann cell behavior in peripheral nerve.

Animals↗
PubMed1995

FORSE-1: a positionally regulated epitope in the developing rat central nervous system.

We designed a protocol to identify cell surface molecules expressed in restricted spatial patterns in the developing central nervous system (CNS) that might be regulated by regionally restricted transcription factors. The immunogen was a membrane fraction from NT2/D1 embryocarcinoma cells that were induced to differentiate into neurons and upregulate Hox gene expression in response to retinoic acid. One monoclonal antibody (mAb), FORSE-1, specifically labels the rostral rat CNS from the earliest stages. Staining is observed in the rostral but not caudal neural folds of the embryo prior to neural tube closure. Staining is enriched in the forebrain as compared to the rest of the CNS, until E18. Between E11.5 and E13.5, only certain areas of the telencephalon and diencephalon are labeled. Later, up to E17.5, FORSE-1 labeling is specifically restricted to the telencephalon, where a correlation with mitotic activity is apparent: the ventricular zone labels with FORSE-1, while the cortical plate is negative. The staining of the neuroepithelium is intensified by acetone fixation, which also reveals, between E11.5 and E13.5, a dorsoventrally restricted, FORSE-1-positive region of the spinal cord. After E18, the entire CNS is labeled, through adulthood. The mAb labels the surfaces of dissociated, living cells. Other, non-CNS areas of FORSE-1 labeling are nasal and otic placodes, nasal epithelium, nasal glands, and early (E9.5-10.5) endoderm. mAb FORSE-1 recognizes an epitope present on both a high-molecular-weight (> 200 kDa) proteoglycan from embryonic and early postnatal brain, and on a 80 kDa doublet that is restricted to the CNS in the adult. These findings suggest the FORSE-1 antigen as a candidate cell surface molecule for mediating regional specification from the earliest stages of CNS development.

Acetone↗
PubMed1995

CD9, a major platelet cell surface glycoprotein, is a ROCA antigen and is expressed in the nervous system.

We previously generated a monoclonal antibody (mAb), ROCA1, which binds preferentially to rostral versus caudal sympathetic ganglia and intercostal nerves. Two other mAbs, ROCA2 and B2C11, bind to the same structures but not in rostrocaudal gradients. All three mAbs recognize a 26 kDa cell surface protein. Amino acid sequence data obtained from the affinity purified 26 kDa protein showed some homology with human CD9, a tetraspan protein implicated in intercellular signaling in hematopoietic cells. Using the PCR, we obtained cDNA clones representing the entire rat CD9 coding sequence from sciatic nerve and sympathetic ganglia. ROCA1, ROCA2, and B2C11 each immunoprecipitate a 26 kDa protein from CHO cells stably transfected with one of the clones, demonstrating that the ROCA cell surface antigen is indeed rat CD9. We find that CD9 mRNA is widely expressed, with particularly high levels present in a number of neural tissues. In situ hybridization demonstrates that peripheral neurons and Schwann cells, as well as adrenal chromaffin cells express CD9 mRNA. Consistent with immunoblot analyses showing that, unlike the ROCA1 epitope, the 26 kDa protein is not expressed in a rostrocaudal gradient, we find similar levels of rat CD9 mRNA in rostral and caudal intercostal nerves. In developing postnatal rat sciatic nerve, CD9 mRNA levels are coordinately regulated with the expression of myelin genes. These results provide another example of a cell surface protein expressed by both hematopoietic and neural cells, and suggest a role for CD9 in intercellular signaling in the nervous system.

Aging↗
PubMed1995

An anti-CD9 monoclonal antibody promotes adhesion and induces proliferation of Schwann cells in vitro.

We have recently found that CD9, a cell surface glycoprotein involved in intercellular signaling in hematopoietic cells, is also expressed by neurons and glia in the peripheral nervous system. Antibody perturbation experiments were conducted to examine the function of CD9 in neural cells. Three anti-CD9 monoclonal antibodies (mAbs) (ROCA1, ROCA2, B2C11) were tested for their ability to promote adhesion of several Schwann cell lines (S-16, RN22, JS1), primary Schwann cells and PC12 cells. Only B2C11 promotes adhesion in all cells tested. Although ROCA2 immunolabels living cells strongly, it had no effect on the adhesion of any of these cells. In addition, ROCA1 and several positive-staining, control mAbs also had no effect. Another mAb, 192-IgG, directed against the low affinity NGF receptor, also promotes the adhesion of S-16, PC12, and primary Schwann cells. In addition to adhesion, contact of S-16 Schwann cells with B2C11 specifically induces morphological changes and robust proliferation. None of the other mAbs, including 192-IgG, induce proliferation of S-16 cells. These results provide evidence that CD9 may be involved in signaling, activation and growth regulation of cells in the nervous system.

Animals↗