Expression of synaptic membrane proteins in gerbil pinealocytes in primary culture

Redecker, P.; Pabst, Heike; Gebert, Andreas; Steinlechner, Stephan

doi:10.1002/(sici)1097-4547(19970301)47:5<509::aid-jnr6>3.0.co;2-l

Cited by 16 publications

(13 citation statements)

References 36 publications

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“…Details and further references concerning these antibodies can be found in previous studies of synaptophysin (Navone et al 1986;Redecker and Bargsten 1993), synaptobrevin II (Redecker 1996;Redecker et al 1997), SNAP-25 (Redecker et al 1996b(Redecker et al , 1997, munc-18-1 , and syntaxin I (Redecker 1996;Redecker et al 1997; an additional monoclonal antibody against syntaxin I, which had not been used in the aforementioned studies, was obtained from Synaptic Systems, Göttingen, Germany, code no. 110 101).…”

Section: Antibodiesmentioning

confidence: 99%

“…Before being immunostained, paraffin sections were subjected to microwave heating in 0.01 M sodium citrate buffer (pH 6.0) for 10 min at a power setting of 900 W. Sections were then processed either for the avidin-biotin-peroxidase complex (ABC) technique or for immunofluorescence staining as outlined previously (Redecker and Bargsten 1993;Redecker et al 1995Redecker et al , 1997Redecker 1996). Briefly, the ABC method included incubation with primary antibodies diluted in phosphate-buffered saline (PBS; anti-synaptophysin 1:2000, anti-synaptobrevin 1:1000, anti-SNAP-25 1:3000, anti-munc-18 1:200, anti-syntaxin 1:1000, antidynamin 1:500, anti-S-antigen 1:800, anti-vimentin 1:200, antilaminin 1:1000, and anti-Ki-67 1:100) containing 0.1% Triton X-100 for 24 h at 4°C, followed by incubation for 30 min at room temperature with the second antibody, viz., biotin-labeled goat antirabbit or anti-mouse IgG (Jackson Immuno Research, West Grove, USA, code nos.…”

Section: Immunohistochemical Protocolmentioning

confidence: 99%

“…The absence of cross-reactivity of the secondary antibodies was assured by using preadsorbed secondary antibodies tailored for multiple labeling procedures and was additionally tested by omitting one of the two primary antibodies. Mounted sections were examined and photographed with a Leitz Orthoplan photomicroscope equipped with fluorescence filters for the detection of Cy2 or Cy3 fluorescence (Redecker et al 1997). To improve the prints of color micrographs, images were digitized and their contrast and brightness were adjusted by means of Adobe Photoshop software.…”

Section: Immunohistochemical Protocolmentioning

confidence: 99%

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Expression of synaptic vesicle trafficking proteins in the developing rat pineal gland

Redecker

2000

Cell and Tissue Research

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Adult mammalian pinealocytes contain several synaptic membrane proteins that are probably involved in the regulation of targeting and exocytosis of synaptic-like microvesicles (SLMVs). Immunohistochemical techniques have now demonstrated the spatiotemporal expression pattern of some of these proteins during rat pineal ontogenesis. Various synaptic vesicle trafficking proteins are detectable in proliferating epithelial cells of the pineal anlage even at embryonic day 17.5 (E 17.5), with the exception of syntaxin I (weakly expressed from E 19.5) and dynamin I (whose levels increase markedly during the first postnatal week). Numerous cells exhibiting strong immunoreactivity for synaptobrevin II, SNAP-25, synaptophysin, and munc-18-1 are distributed throughout the increasingly compact gland at E 19.5 and E 20.5; however, their number declines toward the proximal deep part of the organ. Groups of postmitotic cells situated at the surface of the developing gland exhibit marked immunoreactivity for the aforementioned proteins and lie close to the laminin-immunoreactive outer limiting basement membrane or to its remnants in regions of basement membrane dissolution. We also show that synthesis of vimentin and S-antigen seems to begin earlier during pineal development than previously recognized. Thus, synaptic vesicle trafficking proteins are the earliest molecular markers of pinealocyte differentiation known to date, being expressed well before the onset of rhythmic hormone secretion in the pineal gland, where they may play a role in morphogenetic events. Components of the extracellular matrix such as laminin may be critically involved in the upregulation of synaptic membrane protein expression. The dynamin immunostaining pattern indicates that SLMVs of pinealocytes begin to undergo regulated cycles of exo/endocytosis during postnatal week 1.

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Section: Antibodiesmentioning

confidence: 99%

Section: Immunohistochemical Protocolmentioning

confidence: 99%

Section: Immunohistochemical Protocolmentioning

confidence: 99%

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Expression of synaptic vesicle trafficking proteins in the developing rat pineal gland

Redecker

2000

Cell and Tissue Research

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“…The process terminals may form a gap junction-like structure to the neighboring pinealocytes or glial cells (Fig. 5) (Saez et al, 1991;Redecker et al, 1997). This suggests that the synapse-like structure is the site for exocytosis of MVs for glutamate-mediated intercellular signal transduction.…”

Section: Presence Of Sv2b In Other Neuroendocrine Cellsmentioning

confidence: 99%

“…Pinealocytes express a SNARE complex including an N-ethylmaleimide-sensitive fusion protein and synaptobrevin 2 but are devoid of some proteins found in synaptic vesicles such as synapsins (Moriyama and Yamamoto, 1995b;Moriyama et a!., 1995bMoriyama et a!., , 1996Redecker et al, 1997). The rate of vesicular release is quite slow (around second order in MVs) compared with that in synaptic vesicles (around millisecond order) (Yamada et al, 1996a,b;Yatsushiro et al, 1997).…”

mentioning

confidence: 99%

Synaptic Vesicle Protein SV2B, but Not SV2A, Is Predominantly Expressed and Associated with Microvesicles in Rat Pinealocytes

Hayashi

Yamamoto

Yatsushiro³

et al. 1998

Journal of Neurochemistry

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Abstract:Microvesicles are endocrine counterparts of neuronal synaptic vesicles, and accumulate and secrete classic neurotransmitters. In mammalian pinealocytes, microvesicles accumulate L-glutamate through a vesicular glutamate transporter and secrete it through exocytosis. To characterize the molecular organization of microvesicles in more detail, we investigated in this study the expression and localization of synaptic vesicle protein 2 (SV2) in rat pinealocytes. RT-PCR analysis indicated that transcripts specific for two isoforms, SV2A, a ubiquitous form present in neuronal and endocrine cells, and SV2B, a neuron-specific form, are amplified in pineal RNAs. Northern blotting with specific transcripts indicated that the mRNA for SV2B is predominantly expressed, whereas that for SV2A is below the detection limit. Site-specific antibodies against SV2B recognized a single 72-kDa polypeptide in the pineal membrane fraction, whereas anti-SV2A antibodies did not recognize any polypeptides. Immunohistochemical analysis of cultured cells indicated that SV2B is expressed in pinealocytes but not in other types of cells. SV2B is present in somata and is especially rich in processes, which are filled with microvesicles. SV2B is colocalized with synaptophysin and synaptotagmin, markers for microvesicles. Immunoelectron microscopy indicated that SV2B is associated with microvesicles. These results indicated that SV2B, but not SV2A, is expressed in rat pinealocytes and associated with microvesicles. As SV2B is also expressed in cultured aTC6 clonal pancreatic a cells, SV2B is not a protein specific for neurons. Key Words: SV2B-Expression -Microvesicle (synaptic-like microvesicle) -Pinealocyte-Pineal gland -Synaptic vesicle-aTC6-Endocrine cell.

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Putative neurohemal release zones in the stomatogastric nervous system of decapod crustaceans

Skiebe¹,

Wollenschläger²

2002

J of Comparative Neurology

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The stomatogastric nervous system (STNS) of decapod crustaceans has long been used to study the modulation of small neural circuits. Profiles in the sheath of the nerves and ganglia of the STNS, which contain only dense-core vesicles, have been described in electron microscopical studies (Friend [1976] . These profiles resemble those found in neurohemal organs and suggest the presence of neurohemal release zones in the STNS. To map these putative neurohemal release zones, a combination of two antibodies was used in the present study. A synapsin antibody recognizing vesicle proteins of clear vesicles was combined with a synaptotagmin antibody recognizing vesicle proteins of clear and dense-core vesicles. Exclusive synaptotagmin-like staining, therefore, indicated the regions with only dense-core vesicles. Such a staining was found in a mesh in the perineural sheath of nerves in the STNS of all three species investigated. In the crayfish Cherax destructor and the lobster Homarus americanus, the stained mesh was located in the sheath of nerves connecting all four ganglia of the STNS, whereas in the crab Cancer pagurus it was found on different nerves, which are more directly exposed to the hemolymph in this species. Exclusive synaptotagmin-like staining was also found in a putative neurohemal release zone in the sheath of the circumoesophageal connectives and the postoesophageal commissure in C. destructor. These data suggest that an important source of modulation of the networks and the muscles of the stomach is a compartmentalized release of neurohormones from zones in the STNS.

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Expression of synaptic membrane proteins in gerbil pinealocytes in primary culture

Cited by 16 publications

References 36 publications

Expression of synaptic vesicle trafficking proteins in the developing rat pineal gland

Expression of synaptic vesicle trafficking proteins in the developing rat pineal gland

Synaptic Vesicle Protein SV2B, but Not SV2A, Is Predominantly Expressed and Associated with Microvesicles in Rat Pinealocytes

Putative neurohemal release zones in the stomatogastric nervous system of decapod crustaceans

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