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Biomedical subjects

M Chalfie

Publications and source records attributed to M Chalfie.

At least 37 records · Page 2Linked to original sources
PubMed1995

A mutated acetylcholine receptor subunit causes neuronal degeneration in C. elegans.

Neurotoxicity through abnormal activation of membrane channels is a potential cause of neurodegenerative disease. Here we show that a gain-of-function mutation, deg.3(u662), leads to the degeneration of a small set of neurons in the nematode C. elegans. The deg.3 gene encodes a nicotinic acetylcholine receptor alpha subunit, which in the region of transmembrane domain II is most similar to the neuronal alpha 7 subunits from rat and chicken. The u662 mutation changes a residue in the second transmembrane domain, the domain thought to form the channel pore. A similar change in the equivalent amino acid in the chick protein produces channels that desensitize slowly. Channel hyperactivity may underlie the degenerations seen in the C. elegans deg.3(u662) mutants, since antagonists of nicotinic acetylcholine receptors suppress the deg-3(u662) mutant phenotypes.

Amino Acid Sequence↗
PubMed1995

Control of neuronal development in Caenorhabditis elegans.

Recent research into the development of the nervous system of the nematode Caenorhabditis elegans indicates the importance of multiple cell interactions and combinatorial gene expression. As many of the genes needed for C. elegans neuronal development have counterparts with similar activities in Drosophila melanogaster, the mechanisms of cell specification may be broadly conserved.

Animals↗
PubMed1995

The differentiation and function of the touch receptor neurons of Caenorhabditis elegans.

We have identified several genes required for four aspects of the differentiation and function of a set of six touch receptor neurons in the nematode Caenorhabditis elegans: (1) the generation of appropriate cells; (2) the specification of those cells to differentiate as touch receptors; (3) the maintenance of the differentiated state; and (4) the expression of products need for the cell function. Three major conclusions about the development of the touch cells arise from the analysis of these genes. First, specification of cell fate is a combinatorial process. At least seven genes, none of which are expressed solely in these cells, are needed to restrict the expression of touch-cell features in the appropriate cells. Second, the differentiated state must also be maintained. Three genes appear necessary for this maintenance function. Third, regulation of development is not strictly linear; at least one gene is needed at more than one stage of differentiation. In addition to being interested in the factor that determine cell fate, we are also interested in understanding the molecular basis of mechanosensory transduction. The function class genes are particularly important in this regard, especially those that when mutant result in the loss of the touch response without producing any obvious morphological defects in the touch cells.

Animals↗
PubMed1995

Green fluorescent protein.

Several bioluminescent coelenterates use a secondary fluorescent protein, the green fluorescent protein (GFP), in an energy transfer reaction to produce green light. The most studied of these proteins have been the GFPs from the jellyfish Aequorea victoria and the sea pansy Renilla reniformis. Although the proteins from these organisms are not identical, they are thought to have the same chromophore, which is derived from the primary amino acid sequence of GFP. The differences are thought to be due to changes in the protein environment of the chromophore. Recent interest in these molecules has arisen from the cloning of the Aequorea gfp cDNA and the demonstration that its expression in the absence of other Aequorea proteins results in a fluorescent product. This demonstration indicated that GFP could be used as a marker of gene expression and protein localization in living and fixed tissues. Bacterial, plant and animal (including mammalian) cells all express GFP. The heterologous expression of the gfp cDNA has also meant that it could be mutated to produce proteins with different fluorescent properties. Variants with more intense fluorescence or alterations in the excitation and emission spectra have been produced.

Amino Acid Sequence↗
PubMed1994

Green fluorescent protein as a marker for gene expression.

A complementary DNA for the Aequorea victoria green fluorescent protein (GFP) produces a fluorescent product when expressed in prokaryotic (Escherichia coli) or eukaryotic (Caenorhabditis elegans) cells. Because exogenous substrates and cofactors are not required for this fluorescence, GFP expression can be used to monitor gene expression and protein localization in living organisms.

Animals↗
PubMed1994

Gene interactions affecting mechanosensory transduction in Caenorhabditis elegans.

Genetic screening has identified a group of mec (mechanosensory) genes that are required for the function of a set of six touch-receptor neurons in the nematode Caenorhabditis elegans. Such genes potentially encode components of the mechanosensory apparatus. We have cloned one of these genes, mec-10, which is a member of the degenerin gene family (genes such as mec-4 and deg-1 that can be mutated to cause neurodegeneration). Because components of an amiloride-sensitive sodium channel (alpha, beta and gamma rENaC) from rat share considerable sequence similarity with the C. elegans genes, it is likely that degenerins may function as channel proteins. Here we show that two degenerin homologues (mec-4 and mec-10) are expressed in the same cells, although each provides a unique function. Based on genetic data of mutations affecting mec-10-induced degeneration, we propose that the products of three genes (mec-4, mec-10 and mec-6) form a complex needed for mechanosensation, and that several other mec genes may be important in regulating the putative channel complex.

Amino Acid Sequence↗
PubMed1994

Mutations in the Caenorhabditis elegans beta-tubulin gene mec-7: effects on microtubule assembly and stability and on tubulin autoregulation.

We have sequenced 45 mutations in mec-7, a beta-tubulin gene required for the production of 15-protofilament microtubules in the nematode Caenorhabditis elegans, and have correlated sequence alterations with mutant phenotypes. The expression patterns of most alleles have also been determined by in situ hybridization and immunocytochemistry. Most (12/16) complete loss-of-function alleles, which are recessive, result from nonsense mutations, insertions, or deletions; three others disrupt a putative GTP-binding domain. Three of the four loss-of-function, missense mutations result in elevated mec-7 message levels, suggesting a defect in tubulin autoregulation that may be attributable to a loss in the ability to form heterodimers. Most (8/9) mild alleles are caused by missense mutations. Two mild alleles appear to increase microtubule stability and lead to the elaboration of ectopic neuronal processes in mec-7-expressing cells. Most (15/23) mutations that cause severe dominant or semidominant phenotypes are clustered into three discrete domains; four others occur in putative GTP-binding regions. Many of these dominant mutations appear to completely disrupt microtubule assembly.

Alleles↗
PubMed1993

Cooperative interactions between the Caenorhabditis elegans homeoproteins UNC-86 and MEC-3.

The POU-type homeodomain protein UNC-86 and the LIM-type homeodomain protein MEC-3, which specify neuronal cell fate in the nematode Caenorhabditis elegans, bind cooperatively as a heterodimer to the mec-3 promoter. Heterodimer formation increases DNA binding stability and, therefore, increases DNA binding specificity. The in vivo significance of this heterodimer formation in neuronal differentiation is suggested by (i) a loss-of-function mec-3 mutation whose product in vitro binds DNA well but forms heterodimers with UNC-86 poorly and (ii) a mec-3 mutation with wild-type function whose product binds DNA poorly but forms heterodimers well.

Amino Acid Sequence↗
PubMed1993

Homeobox genes in Caenorhabditis elegans.

It is estimated that approximately 60 homeobox genes occur in the nematode Caenorhabditis elegans. These genes are required for specifying the cell fate of both precursor and terminally differentiated cells. In some cases, highly specific cell functions, such as migration pattern or synaptic connectivity, require the action of these genes.

Animals↗
PubMed1993

Combinatorial control of touch receptor neuron expression in Caenorhabditis elegans.

Six touch receptor neurons with distinctive morphological features sense gentle touch in Caenorhabditis elegans. Previous studies have identified three genes (lin-32, unc-86 and mec-3) that regulate touch cell development. However, since other cell types also require these genes, we suspected that other genes help restrict the expression of touch cell characteristics to the six neurons seen in the wild type. To identify such genes, we have examined mutants defective in genes required for the development of other C. elegans cells for changes in the pattern of touch cell-specific features. Mutations in seven genes either reduce (lin-14) or increase (lin-4, egl-44, egl-46, sem-4, ced-3 and ced-4) the number of touch receptor-like cells. The combinatorial action of these genes, all of which are required for the production of many cell types, restrict the number of cells expressing touch receptor characteristics in wild-type animals by acting as positive and negative regulators and by removing cells by programmed cell death.

Animals↗
PubMed1992

Regulation of the mec-3 gene by the C.elegans homeoproteins UNC-86 and MEC-3.

The mec-3 gene encodes a homeodomain protein with LIM repeats that is required for the specification of touch cell fate in Caenorhabditis elegans. Previous experiments suggested that mec-3 expression requires the product of the unc-86 gene, a POU-type homeoprotein, and mec-3 itself. We have analyzed the control of mec-3 expression by identifying potential cis regulatory elements in the mec-3 gene (by conservation in a related nematode and by DNase I footprinting using unc-86 and mec-3 proteins) and testing their importance by transforming C.elegans with mec-3lacZ fusions in which these sites have been mutagenized in vitro. Both unc-86 and mec-3 proteins bind specifically to the promoter of the mec-3 gene, suggesting that both proteins may be directly involved in the regulation of the mec-3 gene. In addition, the footprint pattern with mec-3 protein is altered in the presence of unc-86 protein. In vivo transformation experiments reveal that some of the binding regions of the two proteins are needed for general positive control and maintenance of mec-3 expression while others have no detectable, unique function. Interestingly, the unc-86 gene appears to be required not only to initiate mec-3 expression but also to maintain it.

Amino Acid Sequence↗
PubMed1992

Developmental and abnormal cell death in C. elegans.

Genetic analysis in Caenorhabditis elegans has identified several genes that function in normal developmental death as well as genes that can mutate to cause inappropriate cell death. The processes whereby some of these abnormal deaths occur depend on genes that participate in normal programmed cell death; others occur by an independent mechanism whereby mutation of members of a gene family leads to cell lysis. Molecular characterization of these 'death' genes in C. elegans is beginning to provide insight into the normal and aberrant mechanisms of cell death.

Animals↗
PubMed1991

The mec-4 gene is a member of a family of Caenorhabditis elegans genes that can mutate to induce neuronal degeneration.

Three dominant mutations of mec-4, a gene needed for mechanosensation, cause the touch-receptor neurons of Caenorhabditis elegans to degenerate. With deg-1, another C. elegans gene that can mutate to induce neuronal degeneration and that is similar in sequence, mec-4 defines a new gene family. Cross-hybridizing sequences are detectable in other species, raising the possibility that degenerative conditions in other organisms may be caused by mutations in similar genes. All three dominant mec-4 mutations affect the same amino acid. Effects of amino-acid substitutions at this position suggest that steric hindrance may induce the degenerative state.

Amino Acid Sequence↗
PubMed1990

The identification and suppression of inherited neurodegeneration in Caenorhabditis elegans.

The dominant mutation deg-1(u38) results in a toxic gene product that leads to the late-onset degeneration of a small number of neurons in the nematode Caenorhabditis elegans. Both intragenic and extragenic mutations as well as changes in wild-type gene dosage can delay or block the time of onset of the neuronal deaths. The deg-1 gene has been cloned and a partial complementary DNA reveals that the gene encodes a novel protein that may act as a membrane receptor. Because the late-onset loss of specific sets of neurons, often as a result of dominant mutations, is characteristic of several human neurodegenerative diseases, the analysis of the deg-1 gene and its suppressors may provide a means of understanding the mechanisms underlying some of these human diseases.

Age Factors↗
PubMed1990

Organogenesis in C. elegans: positioning of neurons and muscles in the egg-laying system.

One of the final stages in the development of egg-laying behavior in the nematode C. elegans is the organization of 8 motor neurons (2 HSN and 6 VC cells) and 8 muscles into a motor system to control the opening of the vulva. Using mutations that disrupt the development of specific components of the egg-laying system and laser microsurgery to ablate selected precursor cells, we have determined that the guidance of the egg-laying neurons and muscles, and in particular the VC neurons and vulval muscles, into the vulval region is dependent on interactions with surrounding epithelial and gonadal tissue and appears to be independent of neuron-neuron and neuron-muscle interactions. The development of the egg-laying system can be described as a series of cell interactions in which certain cells arise through induction and subsequently provide inductive cues themselves.

Animals↗