ABSTRACT
INTRODUCTION: This is the first prospective longitudinal study carried out in a Chinese elderly population with the objective of identifying the incidence and predictors of falls. MATERIALS AND METHODS: This is a population-based cohort study in Hong Kong with 1517 ambulatory elderly Chinese recruited using a multi-stage sampling method. Baseline data on demographic, comorbid diseases, drugs, Activities of Daily Living (ADL) [Barthel Index and Lawton's Instrumental Activities of Daily Living (IADL)], Geriatric Depression Scale (GDS-15), cognitive assessment by the Abbreviated Mental Test (AMT), fear of falling, self-perceived mobility problem, hand grip strength, lower limb power, balance and gait tests were performed. Every subject was followed up for 1 year. RESULTS: Four hundred and one falls occurred in 294 fallers (19.3%) over 1 year of follow-up. The prevalence of falls and recurrent falls were 19.3% and 4.75%, respectively. The incidences of falls (i.e., the fall events) were 220, 324 and 270 per 1000 person-years for men, women and both gender, respectively. The independent predictors of falls were previous history of falls, advancing age, Parkinson's disease, knee extension power and gait speed. The independent predictors of recurrent falls were previous history of falls, self-perceived mobility problem, the knee extension strength and the Total Mobility Score of the Tinetti Balance and Gait Evaluation. CONCLUSIONS: The incidence of falls in the Chinese elderly was 270 per 1000 person-years. History of falls, old age, Parkinson's disease, decreased lower limb power and impairment in balance and gait function were important independent predictors of falls or recurrent falls in the Chinese elderly. Effective fall prevention programmes targeted at improving these risk factors for falls should be developed for the Chinese elderly in Hong Kong and Asia.
Subject(s)
Accidental Falls/statistics & numerical data , Asian People , Geriatric Assessment/methods , Wounds and Injuries/epidemiology , Accidental Falls/prevention & control , Aged , Female , Follow-Up Studies , Gait , Health Status Indicators , Hong Kong/epidemiology , Humans , Incidence , Male , Prevalence , Prospective Studies , Recurrence , Risk Assessment/methods , Risk Factors , Wounds and Injuries/etiologyABSTRACT
PURPOSE: Which functional tests on mobility and balance can better screen older people at risk of falls is unclear. This study aims to compare the Berg Balance Scale (BBS), Tinetti Mobility Score (TMS), Elderly Mobility Scale (EMS) and Timed Up and Go test (TUG) in discriminating fallers from non-fallers in older people. METHOD: This was a case-control study involving one rater who conducted a mobility and balance assessment on subjects using the four functional tests in random sequence. Subjects recruited included 17 and 22 older people with a history of single and multiple falls respectively from a public Falls Clinic, and 39 community-dwellers without fall history and whose age, sex and BMI matched those of the fallers. All subjects underwent the mobility and balance assessment within one day. RESULTS: Single fallers performed better than multiple fallers in all four functional tests but were worse than non-fallers in the BBS, TMS and TUG. The BBS demonstrated the best discriminating ability, with high sensitivity and specificity. The BBS item 'pick up an object from the floor' was the best at screening fallers. CONCLUSION: BBS was the most powerful functional test of the four in discriminating fallers from non-faller.
Subject(s)
Accidental Falls/prevention & control , Activities of Daily Living , Locomotion/physiology , Postural Balance/physiology , Aged , Aged, 80 and over , Analysis of Variance , Case-Control Studies , Confidence Intervals , Disability Evaluation , Female , Humans , Logistic Models , Male , Odds Ratio , ROC Curve , Risk Assessment , Sensitivity and Specificity , Task Performance and AnalysisABSTRACT
Laminin is a major component of all basement membranes. However, its composition varies with location because there are numerous forms of each of the three chains (alpha, beta, and gamma) that together comprise this heterotrimeric molecule. In the neuromuscular system, motor neurons and Schwann cells encounter unique trimers of laminin at different sites. The question thus arises as to whether these local differences in laminin composition act to direct the behavior of these two classes of cells. To address this question, we compared the responses of cultured rat motor neurons and Schwann cells to three forms of rodent laminin purified in our laboratory: Laminin-1 (Lmn-1; alpha1beta1gamma1); Laminin-11 (Lmn-11), a synapse-specific isoform consisting of alpha5beta2gamma1 chains; and a third preparation, a mixture of three kinds of laminin (Lmn-2/4/8), that is enriched for the alpha2, alpha4, beta1, beta2, and gamma1 subunits. Schwann cells attached best to a substrate of Lmn-2/4/8 and showed the weakest adhesion on Lmn-11. Interestingly, no such difference was seen with motor neurons; all three substrates promoted neuronal adhesion, survival, and neurite initiation equally well. With longer time in culture, however, these embryonic motor neurons extended extremely long processes on Lmn-1 and on Lmn-2/4/8, while those on Lmn-11 bore shorter neurites with unusually large, flattened growth cones. These results demonstrate that the behavior of Schwann cells and motor neurons can be regulated directly by the local laminin composition. The precise geometric relationship of these cells at the neuromuscular junction may therefore reflect the unique composition of laminin at this synapse.
Subject(s)
Laminin/analysis , Motor Neurons/physiology , Nerve Tissue Proteins/analysis , Protein Isoforms/analysis , Schwann Cells/physiology , Synapses/chemistry , Animals , Cell Adhesion/physiology , Cell Survival/physiology , Cells, Cultured , Embryo, Mammalian/cytology , Immunohistochemistry , Neurites/chemistry , Neuromuscular Junction/chemistry , Rats , Rats, Sprague-DawleyABSTRACT
CEP-1347, also known as KT7515, a derivative of a natural product indolocarbazole, inhibited motor neuronal death in vitro, inhibited activation of the stress-activated kinase JNK1 (c-jun NH terminal kinase) in cultured spinal motor neurons, but had no effect on the mitogen-activated protein kinase ERK1 in these cells. Results reported here profile the functional activity of CEP-1347/KT7515 in vivo in models of motor neuronal death or dedifferentiation. Application of CEP-1347/KT7515 to the chorioallantoic membrane of embryonic chicks rescued 40% of the lumbar motor neurons that normally die during the developmental period assessed. Peripheral administration of low doses (0.5 and 1 mg/kg daily) of CEP-1347/KT7515 reduced death of motor neurons of the spinal nucleus of the bulbocavernosus in postnatal female rats, with efficacy comparable to testosterone. Strikingly, daily administration of CEP-1347/KT7515 during the 4-day postnatal window of motor neuronal death resulted in persistent long-term motor neuronal survival in adult animals that received no additional CEP-1347/KT7515. In a model of adult motor neuronal dedifferentiation following axotomy, local application of CEP-1347/KT7515 to the transected hypoglossal nerve substantially reduced the loss of choline acetyl transferase immunoreactivity observed 7 days postaxotomy compared to untreated animals. Results from these experiments demonstrate that a small organic molecule that inhibits a signaling pathway associated with stress and injury also reduces neuronal death and degeneration in vivo.
Subject(s)
Apoptosis/drug effects , Axotomy , Carbazoles/pharmacology , Indoles/pharmacology , Mitogen-Activated Protein Kinases , Motor Neurons/drug effects , Motor Neurons/physiology , Animals , Animals, Newborn/physiology , Calcium-Calmodulin-Dependent Protein Kinases/antagonists & inhibitors , Cell Differentiation/drug effects , Chick Embryo , Choline O-Acetyltransferase/metabolism , Enzyme Inhibitors/pharmacology , Female , Hypoglossal Nerve/drug effects , Hypoglossal Nerve/enzymology , Hypoglossal Nerve/pathology , JNK Mitogen-Activated Protein Kinases , Motor Neurons/pathology , Rats , Rats, Sprague-DawleyABSTRACT
Presynaptic and postsynaptic membranes directly oppose each other at chemical synapses, minimizing the delay in transmitting information across the synaptic cleft. Extrasynaptic neuronal surfaces, in contrast, are almost entirely covered by processes from glial cells. The exclusion of glial cells from the synaptic cleft, and the long-term stability of synapses, presumably result in large part from the tight adhesion between presynaptic and postsynaptic elements. Here we show that there is another requirement for synaptic maintenance: glial cells of the skeletal neuromuscular synapse, Schwann cells, are actively inhibited from entering the synaptic cleft between the motor nerve terminal and the muscle fibre. One inhibitory component is laminin 11, a heterotrimeric glycoprotein that is concentrated in the synaptic cleft. Regulation of an inhibitory interaction between glial cells and synaptic cleft components may contribute to synaptic rearrangements, and loss of this inhibition may underlie the loss of synapses that results from injury to the postsynaptic cell.
Subject(s)
Laminin/physiology , Schwann Cells/physiology , Synapses/physiology , Animals , Cells, Cultured , Chick Embryo , Mice , Neurites/physiology , Schwann Cells/ultrastructure , Synapses/ultrastructure , Synaptic Membranes/physiology , Synaptic Membranes/ultrastructure , Tumor Cells, CulturedABSTRACT
PURPOSE: This study was conducted to examine the impact of a change in local alcohol laws on alcohol-related hospital usage. Heavy alcohol consumption and its complications are major health problems in the Arctic. For many years, the sale of alcohol in Barrow was illegal, i.e., "damp" status. In October 1994, the residents of Barrow voted to make both the sale and importation of alcohol illegal, i.e., "dry" status. METHODS: The Public Health Service Hospital in Barrow is the only physician-staffed health care facility for 4,000 residents in Barrow, as well as for five of its seven outlying, dry villages (approximately 2,000 more residents). We retrospectively studied the number of alcohol-related outpatient visits to our hospital from November 1993 through October 1994 (the damp year), and from November 1994 through October 1995 (the dry year). The data used were from the emergency visit log in our outpatient department. RESULTS: We found a 76.5% to 93.2% decrease in alcohol-related outpatient visits when comparing corresponding months during the dry year to those during the damp year. The average overall decrease in alcohol-related outpatient visits was 84.7%. These results were highly significant with a p-value of 0.0022. We conclude that the prohibition of alcohol in remote areas can be an effective method in reducing the amount of alcohol-related health problems and can help make a healthier community.
Subject(s)
Alcohol Drinking/prevention & control , Alcoholism/prevention & control , Outpatient Clinics, Hospital/statistics & numerical data , Alaska/epidemiology , Alcohol Drinking/legislation & jurisprudence , Alcoholism/epidemiology , Arctic Regions , Female , Health Knowledge, Attitudes, Practice , Humans , Male , Retrospective Studies , Seasons , Software , Statistics, NonparametricABSTRACT
CONTEXT: Community availability of alcohol affects alcohol consumption patterns and alcohol-related health and social problems. In Barrow, Alaska, an isolated community at the northernmost reaches of the United States, during a 33-month period, possession and importation of alcohol were legal, completely banned, made legal again, and then banned again. OBJECTIVE: To determine the impact of these public policy changes on alcohol-related outpatient visits at the area hospital. DESIGN: Retrospective review of outpatient records; time-series analysis of alcohol-related visits with respect to community alcohol policy. MAIN OUTCOME MEASURES: Total monthly outpatient visits for alcohol-related problems. RESULTS: There was a substantial decrease in the number of alcohol-related outpatient visits when the ban on possession and importation was imposed compared with baseline. When the ban was lifted, outpatient visits increased; when the ban was reimposed, the number of outpatient visits again decreased. Interrupted time-series analyses confirm that the alcohol ban, its lifting, and its reimposition had a statistically significant and negative effect on the number of alcohol-related outpatient visits (P<.05). CONCLUSION: In a geographically isolated community, the prohibition of alcohol can be an effective public health intervention, reducing the health problems associated with alcohol use.
Subject(s)
Alcohol Drinking/epidemiology , Alcoholic Beverages/supply & distribution , Drug and Narcotic Control/legislation & jurisprudence , Outpatient Clinics, Hospital/statistics & numerical data , Alaska/epidemiology , Alcohol Drinking/adverse effects , Alcoholic Intoxication/epidemiology , Alcoholic Intoxication/prevention & control , Commerce/legislation & jurisprudence , Humans , Models, Statistical , Outpatients/statistics & numerical data , Retrospective StudiesABSTRACT
Choline acetyltransferase (ChAT) is a functional and specific marker gene for neurons such as primary motor neurons that synthesize and release acetylcholine as a neurotransmitter. In adult mammals, transection of the peripheral nerve results in a loss of immunoreactivity for ChAT in the injured motor neurons without affecting their cell number. Using a quantitative RNase protection assay, we have investigated dynamic changes in ChAT mRNA levels following axotomy of motor neurons in the brainstem of adult rats. One week after transection of the left hypoglossal nerve, levels of ChAT mRNA in the ipsilateral side of the hypoglossal motor nucleus decreased dramatically to around 10% when compared to the uninjured contralateral side. When cut axons were chronically exposed to brain-derived neurotrophic factor (BDNF) for 1 week, ChAT mRNA levels were maintained at 63% of control levels. Thus, BDNF can abrogate the injury-induced loss of ChAT mRNA in mature motor neurons in vivo. In contrast, neither neurotrophin 4/5 nor nerve growth factor could prevent the decrease in message. This effect of BDNF on ChAT mRNA levels following peripheral injury to motor neurons demonstrates the existence of regulatory pathways responsive to neurotrophic factors that can "rescue" or "protect" cholinergic gene expression.
Subject(s)
Axons/physiology , Brain-Derived Neurotrophic Factor/pharmacology , Choline O-Acetyltransferase/genetics , Gene Expression Regulation, Enzymologic/drug effects , Motor Neurons/drug effects , RNA, Messenger/metabolism , Animals , Female , Motor Neurons/metabolism , Motor Neurons/ultrastructure , Phosphoglycerate Kinase/genetics , Rats , Rats, Sprague-DawleyABSTRACT
Laminins, heterotrimers of alpha, beta, and gamma chains, are prominent constituents of basal laminae (BLs) throughout the body. Previous studies have shown that laminins affect both myogenesis and synaptogenesis in skeletal muscle. Here we have studied the distribution of the 10 known laminin chains in muscle and peripheral nerve, and assayed the ability of several heterotrimers to affect the outgrowth of motor axons. We show that cultured muscle cells express four different alpha chains (alpha1, alpha2, alpha4, and alpha5), and that developing muscles incorporate all four into BLs. The portion of the muscle's BL that occupies the synaptic cleft contains at least three alpha chains and two beta chains, but each is regulated differently. Initially, the alpha2, alpha4, alpha5, and beta1 chains are present both extrasynaptically and synaptically, whereas beta2 is restricted to synaptic BL from its first appearance. As development proceeds, alpha2 remains broadly distributed, whereas alpha4 and alpha5 are lost from extrasynaptic BL and beta1 from synaptic BL. In adults, alpha4 is restricted to primary synaptic clefts whereas alpha5 is present in both primary and secondary clefts. Thus, adult extrasynaptic BL is rich in laminin 2 (alpha2beta1gamma1), and synaptic BL contains laminins 4 (alpha2beta2gamma1), 9 (alpha4beta2gamma1), and 11 (alpha5beta2gamma1). Likewise, in cultured muscle cells, alpha2 and beta1 are broadly distributed but alpha5 and beta2 are concentrated at acetylcholine receptor-rich "hot spots," even in the absence of nerves. The endoneurial and perineurial BLs of peripheral nerve also contain distinct laminin chains: alpha2, beta1, gamma1, and alpha4, alpha5, beta2, gamma1, respectively. Mutation of the laminin alpha2 or beta2 genes in mice not only leads to loss of the respective chains in both nerve and muscle, but also to coordinate loss and compensatory upregulation of other chains. Notably, loss of beta2 from synaptic BL in beta2(-/-) "knockout" mice is accompanied by loss of alpha5, and decreased levels of alpha2 in dystrophic alpha2(dy/dy) mice are accompanied by compensatory retention of alpha4. Finally, we show that motor axons respond in distinct ways to different laminin heterotrimers: they grow freely between laminin 1 (alpha1beta1gamma1) and laminin 2, fail to cross from laminin 4 to laminin 1, and stop upon contacting laminin 11. The ability of laminin 11 to serve as a stop signal for growing axons explains, in part, axonal behaviors observed at developing and regenerating synapses in vivo.
Subject(s)
Aging/physiology , Laminin/biosynthesis , Muscle, Skeletal/physiology , Neuromuscular Junction/physiology , Neurons/physiology , Peripheral Nerves/physiology , Synapses/physiology , Animals , Cells, Cultured , Chick Embryo , Culture Media, Conditioned , Embryonic and Fetal Development , Ganglia, Parasympathetic/cytology , Ganglia, Parasympathetic/physiology , Gene Expression Regulation, Developmental , Laminin/analysis , Laminin/physiology , Macromolecular Substances , Mice , Mice, Knockout , Motor Neurons/physiology , Muscle Development , Muscle, Skeletal/embryology , Muscle, Skeletal/growth & development , Neurons/cytology , Peripheral Nerves/embryology , Peripheral Nerves/growth & development , Rats , Tumor Cells, CulturedABSTRACT
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.
Subject(s)
Aging/physiology , Axons/physiology , Choline O-Acetyltransferase/analysis , Motor Neurons/cytology , Animals , Cell Survival/physiology , Facial Nerve/physiology , Facial Nerve/ultrastructure , Female , Immunohistochemistry , Male , Mice , Mice, Inbred BALB CABSTRACT
The mutation gly93-->ala of Cu,Zn superoxide dismutase (SOD) is found in patients with familial amyotrophic lateral sclerosis and causes motor neuron disease when expressed in transgenic mice. The progression of clinical and pathological disease was studied in a line of mice designated G1H. Clinical disease started at 91 +/- 14 days of age with fine shaking of the limbs, followed by paralysis and death by 136 +/- 7 days of age. Pathological changes begin by 37 days of age with vacuoles derived from swollen mitochondria accumulating in motor neurons. At the onset of clinical disease (90 days), significant death of somatic motor neurons innervating limb muscles has occurred; mice at end-stage disease (136 days) show up to 50% loss of cervical and lumbar motor neurons. However, neither thoracic nor cranial motor neurons show appreciable loss despite vacuolar changes. Autonomic motor neurons also are not affected. Mice that express wild-type human Cu,Zn SOD remain free of disease, indicating that mutations cause neuron loss by a gain-of-function. Thus, the age-dependent penetrance of motor neuron disease in this transgenic model is due to the gradual accumulation of pathological damage in select populations of cholinergic neurons.
Subject(s)
Aging/physiology , Amyotrophic Lateral Sclerosis/genetics , Amyotrophic Lateral Sclerosis/pathology , Amyotrophic Lateral Sclerosis/physiopathology , Animals , Cell Death , Female , Humans , Male , Mice , Mice, Transgenic , Mutation , Nerve Degeneration , Nerve Regeneration , Superoxide Dismutase/genetics , Vacuoles/ultrastructureABSTRACT
In the adult rat nervous system, motor neurons are recognized specifically by a monoclonal antibody, MO-1. Because binding by MO-1 is lost following axotomy, contact with the target may regulate this motor neuron-specific epitope. To test this hypothesis, we examined the recovery of MO-1 immunoreactivity in hypoglossal neurons following unilateral damage to the hypoglossal nerve. During the first week following nerve crush, neurons in the ipsilateral hypoglossal nucleus lost all immunoreactivity for MO-1. Antibody binding returned with time, and by 4 weeks, 80% of the injured neurons had recovered the MO-1 epitope. Since motor neurons reinnervate their original targets readily following nerve crush, it appears that MO-1 binding is recovered when motor neurons return to their original target muscles in the tongue. When the hypoglossal nerve was cut and inserted into a foreign muscle nearby (the sternomastoid muscle), the MO-1 epitope was not detected in the injured neurons, even when examined 6 weeks after surgery. However, if the sternomastoid muscle was denervated prior to insertion of the hypoglossal nerve, thus allowing the hypoglossal nerve to synapse with this foreign target, increasing numbers of hypoglossal neurons reacquired MO-1 immunoreactivity with time. Our results suggest that the MO-1 epitope is only expressed in motor neurons that are in synaptic contact with skeletal muscle. Thus, a property that distinguishes mature motor neurons from other neuronal phenotypes appears to be regulated by direct synaptic interaction with the postsynaptic target.
Subject(s)
Epitopes , Motor Neurons/immunology , Nerve Regeneration , Animals , Cell Transplantation , Denervation , Female , Hypoglossal Nerve/metabolism , Hypoglossal Nerve/pathology , Hypoglossal Nerve Injuries , Immunohistochemistry , Motor Neurons/transplantation , Muscles/innervation , Muscles/physiology , Nerve Crush , Rats , Rats, Sprague-Dawley , Synapses/physiology , Tongue , Transplantation, HeterotopicABSTRACT
Mutations of human Cu,Zn superoxide dismutase (SOD) are found in about 20 percent of patients with familial amyotrophic lateral sclerosis (ALS). Expression of high levels of human SOD containing a substitution of glycine to alanine at position 93--a change that has little effect on enzyme activity--caused motor neuron disease in transgenic mice. The mice became paralyzed in one or more limbs as a result of motor neuron loss from the spinal cord and died by 5 to 6 months of age. The results show that dominant, gain-of-function mutations in SOD contribute to the pathogenesis of familial ALS.
Subject(s)
Amyotrophic Lateral Sclerosis/genetics , Motor Neuron Disease/genetics , Superoxide Dismutase/genetics , Amyotrophic Lateral Sclerosis/enzymology , Amyotrophic Lateral Sclerosis/pathology , Animals , Brain/enzymology , Disease Models, Animal , Female , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Motor Endplate/pathology , Motor Neuron Disease/enzymology , Motor Neuron Disease/pathology , Motor Neurons/enzymology , Motor Neurons/pathology , Muscles/innervation , Muscles/pathology , Mutation , Pedigree , Spinal Cord/pathology , Superoxide Dismutase/metabolismABSTRACT
The extracellular matrix (ECM) at the neuromuscular junction (NMJ) is biochemically and functionally specialized, and bears molecules that can regulate both the formation and function of this peripheral synapse. We have previously purified one synaptic component of the muscle ECM--a unique laminin isoform named s-laminin--from a rat schwannoma cell line (Chiu et al., 1992). To develop new probes for the ECM, monoclonal antibodies were generated against other components produced by this cell line. One of these new antibodies, 9H6, binds selectively at the synaptic cleft of NMJs in adult rats, but not at extrasynaptic sites on the muscle surface. On Western blots, 9H6 recognizes a 150 kDa band that colocalizes, and copurifies with the laminin-binding, ECM glycoprotein entactin under both reducing and nonreducing conditions. N-terminal sequence analysis also indicates that the 9H6 antigen is related to entactin. However, polyclonal antibodies to entactin stain both synaptic and extrasynaptic sites. Thus, 9H6 appears to identify an entactin epitope with a very restricted distribution. Treatment with N-glycanase reduces the molecular mass of entactin and eliminates 9H6 binding, suggesting that the 9H6 epitope at synapses is dependent on glycosylation. Recent studies have shown that novel isoforms of laminin, collagen IV, agrin, and AChE are selectively sequestered at the NMJ. Our results indicate that the entactin present at the synaptic cleft also differs from entactin present outside the synapse. The synaptic form of entactin may contribute to the unique functions of the ECM at the neuromuscular synapse.
Subject(s)
Epitopes/analysis , Membrane Glycoproteins/analysis , Neuromuscular Junction/chemistry , Neuromuscular Junction/ultrastructure , Amidohydrolases , Animals , Antibodies , Antibodies, Monoclonal , Blotting, Western , Cell Line , Electrophoresis, Polyacrylamide Gel , Extracellular Matrix/chemistry , Extracellular Matrix/ultrastructure , Immunohistochemistry , Kidney/innervation , Membrane Glycoproteins/immunology , Neurilemmoma , Organ Specificity , Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase , Peripheral Nerves/chemistry , Peripheral Nerves/cytology , Rats , Tumor Cells, CulturedABSTRACT
Over the past several years remarkable progress has been made towards unraveling the complex mechanisms that regulate neuronal development. Because the end results of abnormalities in brain development are often developmental disabilities, it is timely to review several recent advances in the field of neurobiology. This review, with contributions from several co-authors, provides a synopsis of breakthroughs from the fields of embryology, cell biology, and molecular genetics that hold promise for exciting clinical application. This article is arranged to reflect the stages of normal development. Understanding the mechanisms underlying neuronal induction, regional specification, neuronal specification, migration, axonal growth, neurotrophic factors, and myelination should clarify the pathophysiology of numerous neurological disorders, and provide new insights into their treatment.
Subject(s)
Cell Differentiation/physiology , Nervous System/cytology , Animals , Axons/physiology , Cell Differentiation/genetics , Cell Movement/genetics , Cell Movement/physiology , Humans , Mutation , Myelin Proteins/genetics , Nerve Fibers, Myelinated/physiology , Nerve Growth Factors , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/physiology , Neurons/cytology , Oligodendroglia/cytology , Oligodendroglia/transplantationABSTRACT
In adult mammals, transection of the hypoglossal nerve results in a dramatic loss of choline acetyltransferase (ChAT) in the hypoglossal motor neurons without affecting their cell number. This injury-induced reduction in ChAT is prevented when brain-derived neurotrophic factor (BDNF) is applied to the proximal end of the transected nerve. In contrast, application of ciliary neurotrophic factor (CNTF) has no such effect, even though both factors are known to rescue developing motor neurons from cell death. These results suggest that BDNF may regulate the phenotypic expression of ChAT in mature motor neurons, and indicate that the sensitivity and response of motor neurons to such neurotrophic agents change with development.
Subject(s)
Axons/physiology , Motor Neurons/drug effects , Nerve Growth Factors/pharmacology , Nerve Tissue Proteins/pharmacology , Animals , Brain-Derived Neurotrophic Factor , Cell Death/drug effects , Choline O-Acetyltransferase/metabolism , Ciliary Neurotrophic Factor , Female , Hypoglossal Nerve/cytology , Hypoglossal Nerve/drug effects , Hypoglossal Nerve/enzymology , Immunohistochemistry , Motor Neurons/enzymology , Motor Neurons/physiology , Phenotype , Rats , Rats, Sprague-Dawley , Tongue/innervationABSTRACT
Somatic motor neurons begin to express the transmitter synthesizing enzyme, choline acetyltransferase (ChAT) and the low-affinity nerve growth factor receptor (NGFR) during embryonic development. However, as motor neurons mature in postnatal life, they lose immunoreactivity for NGFR and acquire a motor neuron-specific epitope that is recognized by the monoclonal antibody, MO-1. The present study was undertaken to examine the effect of nerve injury in adult rats on these three developmentally regulated markers in two populations of somatic motor neurons. Unilateral transection, ligation, or crushing of the sciatic nerve resulted in a loss of MO-1 binding and a concomitant rise in immunoreactivity for NGFR within axotomized motor neurons in lumbar levels of the spinal cord. These changes, detectable within 5 days following nerve injury, are reversed with reinnervation, but persist if reinnervation is prevented by chronic axotomy. Thus, regulation of the expression of NGFR and the MO-1 epitope appears to be critically dependent upon interactions between motor neurons and target muscles. These observations are also consistent with the idea that during regeneration, neurons may revert to a developmentally immature state; in motor neurons, this state is characterized by the presence of NGFRs and the absence of the MO-1 epitope. Transection of the hypoglossal nerve, a purely motor nerve, resulted in a similar loss of MO-1 binding and a selective rise in NGFR immunoreactivity in neurons within the ipsilateral hypoglossal motor nucleus. In addition, immunoreactivity for ChAT was also lost in axotomized hypoglossal motor neurons. In contrast, injury to the sciatic nerve, which bears both sensory and motor axons, did not result in any detectable change in ChAT immunoreactivity in spinal motor neurons.
Subject(s)
Epitopes/immunology , Motor Neurons/immunology , Nerve Tissue Proteins/immunology , Receptors, Nerve Growth Factor/immunology , Animals , Antibodies, Monoclonal/immunology , Axons , Choline O-Acetyltransferase/biosynthesis , Gene Expression Regulation , Hypoglossal Nerve/physiology , Hypoglossal Nerve Injuries , Ligation , Nerve Crush , Nerve Regeneration , Nerve Tissue Proteins/biosynthesis , Rats , Rats, Sprague-Dawley , Receptors, Nerve Growth Factor/biosynthesis , Sciatic Nerve/injuries , Sciatic Nerve/physiology , Spinal Cord/growth & developmentABSTRACT
The extracellular matrix (ECM) at the vertebrate neuromuscular junction is a repository of functionally important molecules, some of which can regulate the formation of synapses during regeneration. One candidate molecule is s-laminin, a 185-kDa homologue of the laminin B1 chain. Whereas several members of the laminin family are present throughout the ECM ensheathing muscle fibers, immunoreactivity for s-laminin is found selectively at synaptic sites in adult and embryonic rats, and is detectable at a time when synaptogenesis is taking place during development. We have reported previously that a rat schwannoma cell line, D6P2T, produces and releases large amounts of s-laminin in culture. We have now purified s-laminin from medium conditioned by these cells by using a simple three-step procedure. Serum-free, conditioned medium is separated by ion-exchange chromatography on DEAE-Sephacel, followed by size-exclusion chromatography on 500 HR-Sephacryl. Finally, s-laminin is dissociated from other ECM components by agarose gel electrophoresis under reducing conditions and recovered in solution by extracting slices of agarose gel. The purified preparation displays one silver-stained band that is recognized by three monoclonal antibodies known to bind to different epitopes on s-laminin. Lectin-binding studies demonstrate that s-laminin is a glycoprotein and bears many of the carbohydrate moieties present on the B1 and B2 chains of laminin. Thus, the three 185-220-kDa members of the laminin family are related in both their protein and carbohydrate domains.
Subject(s)
Laminin/isolation & purification , Lectins/metabolism , Synapses/metabolism , Animals , Blotting, Western , Culture Media , Electrophoresis, Polyacrylamide Gel , Isomerism , Laminin/metabolism , Neurilemmoma/chemistry , Neurilemmoma/pathology , Tumor Cells, CulturedABSTRACT
In order to identify early events in the differentiation of motor neurons, the expression of several developmentally regulated, neuronal molecules was investigated by immunohistochemistry on consecutive sections of cervical spinal cord. Motor neurons are among the first neurons to be born and to differentiate within the embryonic rat spinal cord. They undergo their terminal mitosis on embryonic days 10 and 11 (E10-11) and acquire detectable levels of the transmitter synthesizing enzyme, choline acetyltransferase, by E11.5. Staining with antibodies to the 68 kD neurofilament protein revealed motor neurons extending processes out the ventral root as early as E10.5. Monoclonal antibodies to two different epitopes on the cell adhesive molecule, NCAM, bound to myotomes on E10.5, and began to recognize ventral horn neurons by E11. Two other markers of developing neurons, the growth-associated protein, GAP-43, and the surface glycoprotein, TAG-1, were clearly detected on young motor neurons by E11.5. Thus, during the 36 hours following the final mitosis of their precursors, motor neurons acquire cytoskeletal, enzymatic, and cell surface components that distinguish them from other developing cells within the spinal cord. Not all of the newly acquired molecules continue to be expressed by motor neurons. Immunoreactivity for TAG-1 was lost by E12.5, followed by a gradual reduction of immunoreactivity for GAP-43 and the highly polysialylated form of NCAM. By E15, only antibodies to choline acetyltransferase (Phelps et al., J. Comp. Neurol. 307:1-10, 1990), and to neurofilaments, selectively stained motor neurons within the embryonic spinal cord. The transient presence of GAP-43, TAG-1, and the embryonic form of NCAM coincides with a period of vigorous axonal growth and declines when motor neurons reach their targets. This report describes the temporal sequence of early stages in the differentiation of the rodent motor neuronal phenotype. Some of these changes may be related to interactions with their synaptic partners.
Subject(s)
Motor Neurons/physiology , Neurofilament Proteins/analysis , Spinal Cord/embryology , Animals , Antibodies, Monoclonal , Cell Adhesion Molecules, Neuronal/analysis , Cell Differentiation , Contactin 2 , Embryonic and Fetal Development , Female , GAP-43 Protein , Male , Membrane Glycoproteins/analysis , Molecular Weight , Motor Neurons/cytology , Nerve Tissue Proteins/analysis , Pregnancy , Rats , Rats, Inbred Strains , Spinal Cord/cytology , Spinal Nerve Roots/embryologyABSTRACT
Components of the extracellular matrix (ECM) have been implicated in the regulation of neuronal migration, axonal growth, and synaptogenesis. We have examined cultures of glial cells, Schwann cells, and schwannomas for the expression of two components of the ECM, laminin and s-laminin, using immunohistochemical and Western blot techniques. Laminin is a potent promotor of neurite outgrowth in cultures of both central and peripheral neurons, and is present in all ECMs. In contrast, s-laminin (for synaptic laminin), a recently described homolog of laminin, is highly localized at the neuromuscular synaptic cleft (Sanes and Chiu, Cold Spring Harbor Symp. Quant. Biol. 1983;48:667-678; Chiu and Sanes, Dev. Biol. 1984;103:456-467) and shows selective adhesivity for motor neurons (Hunter et al. Cell 1989;59:905-913). While the distribution of these ECM components have been well documented in situ, the sources of these extracellular molecules are unclear. We report that astrocytes cultured in serum-free medium maintain an organized ECM that only bears laminin immunoreactivity; s-laminin appears to be sequestered intracellularly. However, both molecules are found in the astrocyte conditioned medium. Thus, under these growth conditions, astrocytes produce and release laminin and s-laminin, but only incorporate the former into an ECM. In contrast, neither molecule is present in comparable cultures of oligodendrocytes. Although no established ECM is seen in cultures of Schwann cells or schwannomas, laminin and s-laminin immunoreactivity are present within cells and in the conditioned media. These results indicate that certain populations of non-neuronal support cells and cell lines can produce and release both synaptic and extrasynaptic components of the ECM. The assembly of these different molecules into an organized basal lamina may require the presence of additional factors or interaction with neurons.