Biomedical subjects

Esther Asan

Publications and source records attributed to Esther Asan.

10 recordsLinked to original sources

PubMed2006

Bruchpilot, a protein with homology to ELKS/CAST, is required for structural integrity and function of synaptic active zones in Drosophila.

Neurotransmitters are released at presynaptic active zones (AZs). In the fly Drosophila, monoclonal antibody (MAB) nc82 specifically labels AZs. We employ nc82 to identify Bruchpilot protein (BRP) as a previously unknown AZ component. BRP shows homology to human AZ protein ELKS/CAST/ERC, which binds RIM1 in a complex with Bassoon and Munc13-1. The C terminus of BRP displays structural similarities to multifunctional cytoskeletal proteins. During development, transcription of the bruchpilot locus (brp) coincides with neuronal differentiation. Panneural reduction of BRP expression by RNAi constructs permits a first functional characterization of this large AZ protein: larvae show reduced evoked but normal spontaneous transmission at neuromuscular junctions. In adults, we observe loss of T bars at active zones, absence of synaptic components in electroretinogram, locomotor inactivity, and unstable flight (hence "bruchpilot"-crash pilot). We propose that BRP is critical for intact AZ structure and normal-evoked neurotransmitter release at chemical synapses of Drosophila.

Adaptor Proteins, Signal Transducing↗

PubMed2005

"Subcellular proteomics" of neuromelanin granules isolated from the human brain.

"Subcellular proteomics" is currently the most effective approach to characterize subcellular compartments. Based on the powerful combination of subcellular fractionation and protein identification by LC-MS/MS we were able for the first time to 1) isolate intact neuromelanin granules from the human brain and 2) establish the first protein profile of these granules. This compartment containing neuromelanin (NM) is primarily located in the primate's substantia nigra, one of the main brain regions that severely degenerates in Parkinson disease. We used mechanic tissue disaggregation, discontinuous sucrose gradient centrifugation, cell disruption, and organelle separation to isolate NM granules from human substantia nigra. Using transmission electron microscopy we demonstrated that the morphological characteristics of the isolated NM granules are similar to those described in human brain tissue. Fundamentally we found numerous proteins definitely demonstrating a close relationship of NM-containing granules with lysosomes or lysosome-related organelles originating from the endosome-lysosome lineage. Intriguingly we further revealed the presence of endoplasmic reticulum-derived chaperones, especially the transmembrane protein calnexin, which recently has been located in lysosome-related melanosomes and has been suggested to be a melanogenic chaperone.

Calnexin↗

PubMed2005

Glial and neuronal expression of polyglutamine proteins induce behavioral changes and aggregate formation in Drosophila.

Patients with polyglutamine expansion diseases, like Huntington's disease or several spinocerebellar ataxias, first present with neurological symptoms that can occur in the absence of neurodegeneration. Behavioral symptoms thus appear to be caused by neuronal dysfunction, rather than cell death. Pathogenesis in polyglutamine expansion diseases is largely viewed as a cell-autonomous process in neurons. It is likely, however, that this process is influenced by changes in glial physiology and, at least in the case of DRPLA glial inclusions and glial cell death, seems to be an important part in the pathogenesis. To investigate these aspects in a Drosophila model system, we expressed polyglutamine proteins in the adult nervous system. Glial-specific expression of a polyglutamine (Q)-expanded (n=78) and also a nonexpanded (n=27) truncated version of human ataxin-3 led to the formation of protein aggregates and glial cell death. Behavioral changes were observed prior to cell death. This reveals that glia is susceptible to the toxic action of polyglutamine proteins. Neuronal expression of the same constructs resulted in behavioral changes similar to those resulting from glial expression but did not cause neurodegeneration. Behavioral deficits were selective and affected two analyzed fly behaviors differently. Both glial and neuronal aggregates of Q78 and Q27 appeared early in pathogenesis and, at the electron microscopic resolution, had a fibrillary substructure. This shows that a nonexpanded stretch can cause similar histological and behavioral symptoms as the expanded stretch, however, with a significant delay.

Age Factors↗

PubMed2005

Motoneuron survival after C7 nerve root avulsion and replantation in the adult rabbit: effects of local ciliary neurotrophic factor and brain-derived neurotrophic factor application.

BACKGROUND: The authors investigated the extent and time course of motoneuron cell death after C7 ventral nerve root avulsion under conditions resembling the trauma mechanism in clinical situations. In addition, they evaluated the effect on motoneuron survival of locally applied ciliary neurotrophic factor and brain-derived neurotrophic factor, with the aim of improving preconditions for successful regeneration of peripheral motor innervation. METHODS: Forty-four New Zealand White rabbits were operated on using a dorsal approach. The dorsal spinal nerve roots of segment C7 were cut, and the ventral roots were completely pulled out from the spinal cord. In seven experimental groups, ciliary neurotrophic factor, brain-derived neurotrophic factor, or both were applied to the lesion site using different application methods and compared with two control groups. One or 3 weeks after the operation, the animals were euthanized and segments C6 to C8 were studied histologically. In group 9, the avulsed rootlets were replanted into the ventrolateral spinal cord and the effect of replantation on motoneuron survival was assessed at 3 weeks postoperatively. RESULTS: The results indicated that within a period of 7 days, 54.4 +/- 12.1 percent of the motoneurons in segments C6 to C8 died without any therapy. Local application of ciliary neurotrophic factor or brain-derived neurotrophic factor lowered motoneuron loss significantly to 16.9 +/- 14.3 percent and 28.0 +/- 11.4 percent, respectively (p < 0.05). The reduction in motoneuron loss persisted after 3 weeks' survival time (23.1 +/- 4.3 percent in ciliary neurotrophic factor-treated animals, and 22.3 +/- 8.4 percent in brain-derived neurotrophic factor-treated animals, p < 0.05). Survival rates were not significantly higher after treatment with a combination of both factors (motoneuron loss, 33.5 +/- 7.1 percent). CONCLUSION: The authors conclude that the early application of neurotrophic factors appears to be a promising technique to improve motoneuron survival after nerve root avulsion.

Animals↗

PubMed2004

News and views in Histochemistry and Cell Biology.

Advances in histochemical methodology and ingenious applications of novel and improved methods continue to confirm the standing of morphological means and approaches in research efforts, and contribute significantly to increasing our knowledge about structures and functions in all areas of the life sciences from cell biology to pathology. Reports published during recent months documenting this progress are summarized in the present review.

Animals↗

PubMed2003

Innovative techniques and applications in histochemistry and cell biology.

Recent studies documenting novel histochemical methods and applications in cell biology and in other areas of the life sciences have again rendered insights into structure and functions of tissues, cells, and cellular components to the level of proteins and genes. Particularly, sophisticated microscopic techniques have proved to be able to significantly advance our knowledge. Findings of recent investigations representing this progress are summarized in the present review.

Animals↗

PubMed2003

Interrelations between monoaminergic afferents and corticotropin-releasing factor-immunoreactive neurons in the rat central amygdaloid nucleus: ultrastructural evidence for dopaminergic control of amygdaloid stress systems.

Ample evidence implicates corticotropin-releasing factor (CRF)-producing neurons of the central amygdaloid nucleus (CeA) in vegetative, endocrine, and behavioral responses to stress and anxiety in laboratory rats. Monoaminergic systems are involved in modulating these responses. In the present paper, interrelations between CRF-immunoreactive (ir) neurons, and noradrenergic, serotonergic, and dopaminergic afferents were studied using single and double immunolabeling for light and electron microscopy in the rat CeA. Dopaminergic axons formed dense plexus in the CeA overlapping with the localization of CRF-ir neurons, and their terminals formed frequent associations with CRF-ir somata. Contacts of serotonergic axons on CRF-ir neurons were few, and contacts of noradrenergic axons were the exception. Ultrastructurally, symmetric synapses of dopaminergic terminals on CRF-ir somata and dendrites were found. More than 83% of CRF-ir somata were contacted in single ultrathin sections. About half of these possessed two or more contacts. Of non-ir somata, 37% were contacted by dopaminergic terminals, and only 13% of these had two or more contacts. Correlative in situ hybridization indicated that CeA CRF-ir neurons may express receptor subtype dopamine receptor subtype 2. In conclusion, dopaminergic afferents appear to specifically target CeA CRF neurons. They are thus in a position to exert significant influence on the rat amygdaloid CRF stress system.

Amygdala↗

PubMed2002

Progress in focus: recent advances in histochemistry and cell biology.

Advances in histochemical and cell biological techniques enable increasingly refined investigations into the cellular and subcellular distribution of specific molecules and into their role in dynamic processes; thus progress in these fields complements the growing knowledge in genomics and proteomics. The present review summarizes recent technical progress and novel applications.

Animals↗

PubMed2002

A mouse model for Sorsby fundus dystrophy.

PURPOSE: Sorsby fundus dystrophy (SFD) is a rare, late-onset macular dystrophy caused by mutations in the tissue inhibitor of metalloproteinases-3 (TIMP3) gene. The known mutations introduce potentially unpaired cysteine residues in the C terminus of the protein and result in the formation of higher-molecular-weight protein complexes of as yet unknown composition and functional consequences in the pathologic course of SFD. To facilitate in vivo investigation of mutant TIMP3, the authors generated a knock-in mouse carrying a disease-related Ser156Cys mutation in the orthologous murine Timp3 gene. METHODS: Site-directed mutagenesis and homologous recombination in embryonic stem (ES) cells was used to generate mutant ES cells carrying the Timp3(S156C) allele. Chimeric animals were obtained, of which two displayed germline transmission of the mutated allele. Molecular genetic, biochemical, electron microscopic, and electrodiagnostic techniques were used for characterization. RESULTS: At 8 months of age, knock-in mice showed abnormalities in the inner aspect of Bruch's membrane and in the organization of the adjacent basal microvilli of the retinal pigment epithelium (RPE). Changes resembling those in the mutant animals were also present to some extent in normal littermates, but only at an advanced age of 30 months. Long-term electrodiagnostic recordings indicated normal retinal function throughout life. The biochemical characteristics of the mutant protein appear similar in humans and knock-in mice, suggesting common molecular pathways in the two species. The localization of the mutant protein in the eye is normal, although there is evidence of increased Timp3 levels in Bruch's membrane of mutant animals. CONCLUSIONS: The knock-in mice display early features of age-related changes in Bruch's membrane and the RPE that may represent the primary clinical manifestations of SFD. In addition, our immunolabeling studies and biochemical data support a model proposing that site-specific excess rather than absence or deficiency of functional Timp3 may be the primary consequence of the known Timp3 mutations.

Animals↗

PubMed1999

Chemosensory Perception in the Gut.

The ability of the gut mucosa to sense the chemical composition of chyme is important for gastrointestinal functions. The demonstration of gustducin and transducin, two alpha-subunits of GTP-binding proteins involved in gustatory signal transduction, in gastrointestinal epithelial cells provides first clues to the molecular basis of enteric chemosensitivity. Nitric oxide may play a role as a secondary messenger.

Journal Article↗