Protein expression patterns of identified neurons and of sprouting cells from the leech central nervous system

Wu, Xueqing; Ritter, Barbara; Schlattjan, Jan Henrik; Leßmann, Volkmar; Heumann, Rolf; Dietzel, Irmgard D.

doi:10.1002/1097-4695(20000905)44:3<320::aid-neu3>3.0.co;2-4

Cited by 7 publications

(1 citation statement)

References 53 publications

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“…After rehydration, IEF voltage was raised gradually to 8000 V and gels were run at 207C until reaching 180 000 V?h. After focusing, proteins were reduced and subsequently carbamidomethylated with 130 mM DTT and with 135 mM iodoacetamide, respectively, in 6 M urea, 375 mM Tris-HCl, 2% SDS, 20% glycerol, 0.02% CBB, pH 8.8 [20]. Second dimensions were run on 12% polyacrylamide/0.8% piperazine diacrylamide in Tris/Tricine buffer as described by Schägger and von Jagow [21].…”

Section: -Dementioning

confidence: 99%

Proteome modifications of the medicinal leech nervous system under bacterial challenge

Vergote¹,

Macagno²,

Sautière³

2006

Proteomics

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Once considered as lacking intrinsic immune mechanisms, the CNS of vertebrates is now known to be capable of mounting its own innate immune response. Interestingly, while invertebrates have been very useful in the interpretation of general vertebrate innate immunity mechanisms, only scarce data are available on the immune response of nervous tissue within this group. This study provides new data on the innate immune response of medicinal leech Hirudo medicinalis CNS. We identified several spots in 2-D gels of leech CNS proteins that showed specific changes following bacterial challenge, thus demonstrating the ability of the leech nervous system to mount a response to an immune stress. Protein identifications were based on comparison of sequence data with publicly available databases and a recently established leech ESTs database. The broad nature of the identified proteins suggests a clear involvement of cytoskeletal rearrangements, endoplasmic reticulum stress, modulation of synaptic activity and calcium mobilization, all during the first 24 hours of this response. Moreover, several of these proteins are specifically expressed in glial cells, suggesting an important role for glial cells in the immune response of the leech nervous system, similar to what has been observed in vertebrates.

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Section: -Dementioning

confidence: 99%

Proteome modifications of the medicinal leech nervous system under bacterial challenge

Vergote¹,

Macagno²,

Sautière³

2006

Proteomics

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Extracellular matrix glycoproteins inhibit neurite production by cultured neurons

De‐Miguel

Vargas

Arias³

et al. 2002

J of Comparative Neurology

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We have analyzed the role of extracellular matrix glycoproteins in the formation of a bipolar outgrowth pattern of identified leech neurons in culture. Adult anterior pagoda (AP) neurons cultured on the inner surface of the ganglion capsules that surround central nervous system, generate two processes oriented in opposite directions. This pattern differs from those produced by these neurons cultured on other substrates, and is similar to the pattern of developing AP neurons at embryonic day 10. We used different lectins to identify subsets of glycoproteins in the extracellular matrix (ECM) of the capsules and to study their contribution to the formation of the bipolar outgrowth pattern. ECM glycoproteins binding to peanut agglutinin (PNA) or Galanthus nivalis aglutinin (GNA) lectins were detected in ganglion capsules and in ganglion extracts that had been separated by electrophoresis and blotted to nitrocellulose membranes. Four protein bands bound to PNA lectin and six other bands, including laminin subunits, bound to GNA lectin. Other lectins failed to recognize any of the proteins. For AP neurons cultured on capsules, addition of PNA lectin to the culture medium produced a dose-dependent increase in the number of primary neurites without affecting their shape, length or number of branch points. However, PNA lectin used as substrate did not affect sprouting of AP neurons. Our results suggest that PNA-binding extracellular matrix glycoproteins regulate the formation of the bipolar pattern of AP neurons by inhibiting the formation of neurites.

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Steps in the formation of a bipolar outgrowth pattern by cultured neurons, and their substrate dependence

De‐Miguel

Vargas

2002

J. Neurobiol.

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We studied the steps in the formation of the bipolar outgrowth pattern of cultured adult Anterior Pagoda (AP) neurons of the leech growing on a central nervous system (CNS) homogenate as substrate. This pattern, which consists of two primary neurites directed in opposite directions plus some bifurcations, resembles their embryonic pattern but is different from the patterns they develop in culture on leech laminin or Concanavalin A as substrates. In eight neurons that were studied, one primary neurite formed and branched several hours before the second one. Time-lapse video analysis showed that between 12 and 36 h of growth, the more proximal branch of the early neurite migrated retrogradely, rotated, and formed the second primary branch. Both neurites elongated until the total neurite length reached 130-160 microm, when the elongation of primary neurites became synchronous with the retraction of secondary processes, suggesting competition. The substrate dependence of these events was tested by plating AP neurons on leech laminin. On this substrate AP neurons produced multiple independent primary neurites with branches. Retraction of some large branches was followed by their regrowth, and did not correlate with the changes in other neurites. We propose that the dynamics in the formation of the bipolar outgrowth pattern of AP neurons arise from inhibitory extracellular matrix molecules, which reduce the synthesis of precursors for neurite formation.

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Protein expression patterns of identified neurons and of sprouting cells from the leech central nervous system

Cited by 7 publications

References 53 publications

Proteome modifications of the medicinal leech nervous system under bacterial challenge

Proteome modifications of the medicinal leech nervous system under bacterial challenge

Extracellular matrix glycoproteins inhibit neurite production by cultured neurons

Steps in the formation of a bipolar outgrowth pattern by cultured neurons, and their substrate dependence

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