Beruflich Dokumente
Kultur Dokumente
SESSION 2015-16
SWAGATA BOSE
Roll No-18
CLASSIFICATION
Kingdom- Animalia
Phylum- Nematoda
Class- Chromadorea
Order- Rhabditida
Family- Rhabditidae
Genus- Caenorhabditis
Species- elegans
INTRODUCTION-
In the lab, it is grown by spreading a layer of E. coli onto a plate and letting the animal feed on the
bacterial lawn.
It was considered as the only organisms to have its complete “ connectome” (2012) a neuronal “wiring”
diagram.
They have a short generation about 3 days from egg-laying to adulthood. Its brood size is less than 300.
It was first introduced and used by Sydney Brenner (1963) to study development and neurology.
His fundamental contributions, ranging from molecular biology, developmental biology to genomics
played a crucial role in science and technology.
Key contributions made by Sydney Brenner were - Discovery of messenger mRNA and Solving genetic
code.
The new challenge in front of him was the Development of the “Nervous system of the model
organisms, Caenorhabditis elegans”, and the two vital reasons for selecting C.elegans as the model
organism was its simple nervous system consisting only 302 neurons and it was also amenable to
genetic analysis.
It has 100 Mb genome which is fully sequenced and is equal to 19000 genes. They play a vital role in the
study of gene expression.
It has a very short life span of about 2-6 days which wholly depends on the temperature( About 50⁰C-2
days, 20⁰C-3 days, 06⁰C-5 days).
It is a self – fertilize (hermaphrodite) organism, which produces 300-350 offsprings. Its fertilization
occur with true males and number of offsprings may exceed upto 1000 .
Its Post -embryonic cell lineages were determined by Sulston and Horwitz, 1976.
It shows Programmed cell death mechanism which was discovered by Horwitz, 1982.
Its complete embryonic cell lineages determined by Deppe et al, Sulston 1983.
Its complete connectivity of nervous system was established on (1986). The RNAi and miRNA discovered
in worms (1991-98) (Nobel Prize: Fire, Mello 2006).
The first use of GFP in animals(1994)was done (Nobel Prize: Chalfie, 2008)and also the first animal
whose whole genome was sequenced(97Mb, now 100.3Mb).
Similarities between C . elegans and Humans-
Both organisms , development proceeds through :
Both of them are having a nervous system, muscular system which help in movements and a gut for
completion of various metabolic processes. It is capable of performing very simple behaviors.
It produces sperms and eggs and undergo reproduction, although normally as a hermaphrodite in case
of C.elegans. After their reproductive phase , the worms gradually undergo senescence where body
metabolisms slows down and the organism ultimately dies.
This is how a very simple organisms exhibits all the phases of life in a short span of time.
About 70% of human genes have a clear C. elegans homolog (this includes human disease genes).
Human genes can often rescue the worm mutants.
C.elegans are well characterized organisms having biological functions similar to humans.
They are helpful for applying strategies in modern science and medicines to find out different ailments
occur to human.
Its one of the remarkable feature is its reaction with UV light and emit an intense blue fluorescence that
is similar to lysosomes only as lysosomes also have acidic interior and capacity of endocytosis , it also act
as storage organelles.
• Hermaphrodite- self fertile female having 5 pair of autosomes and XX sex chromosome
• True males- only produce sperms ( not used as model organisms) 5 pairs of autosomes and XO
sex chromosome
The 5 autosomes represented as I-V and X (6 total). The chromosomes are non-disjunctional, with one
less chromosome, males develop . They have 79 more neurons and 25 more muscle cells than
hermaphrodites, used for mating.
Hermaphrodite produces many progeny about 300 worms. Adult hermaphrodites have 959 somatic
LIFE CYCLE-
Under favorable environmental condition, the hatched larvae develop through 4 stages or molts (L1 to
L4) .
Rapid life cycle is about 3 days at 20 degrees Celsius from eggs -> L1-L4 -> adults -> eggs.
Offspring: A hermaphrodite can produce about 300-350 offspring under self-fertilization and more if it
mates with males.
The Duaer stage is the “ Alternative life form of C. elegans “ the name was derived from German ( Duaer
- Permanent)
C. elegans enter an alternative third larval stage when, culture plate is over crowded or when food
scarcity is there.
Duaer larvae are stress resistant, thin and mouth is sealed ; hence cannot take in food. They remain
viable for few months (about 3 months). These worms can live 10 times more than their normal life
span.
C. elegans can adopt an alternative life form, called the dauer larval stage, if plates are too crowded or if
food is scarce. Dauer larvae are thin and can move but their mouths are plugged and they cannot eat.
Interestingly, dauers can remain viable for three months. They appear to be non-aging: dauer larvae can
roam around for months and then reenter the L4 stage when they encounter a food source and live
about 15 more days.
At 25ºC a generation time is about 3-4 days, where as it extends to almost a week at 16ºC. Some
mutations in C. elegans are also temperature sensitive, only showing a phenotype at either 16ºC or
25ºC.
C. elegans cell lineage
One amazing advantage of worms is that every worm has the exact same number of somatic cells.
Somatic cells arise by an INVARIANT cell lineage. The divisions are invariant in pattern, timing, and
In the presence of food, cell divisions resume and postembryonic developmental program begins 3
hours after hatching. a first stage larva has approximately 671 cells, 113 undergo programmed death in
the course of development. About 10% of the remaining 558 cells in a newly hatched larva (51 in
hermaphrodites, 55 in the male), are blast cells that divide further.
DRUNKEN WORMS REVEAL A GENETIC BASIS OF ALCOHOL RESPONSE-
Gene responsible for causing alcohol sensitivity discovered by Andrew Davies in June 10, 2004
Subtle differences are seen between worm strains in a gene for a brain protein called NPR-1 (Natriuretic
peptide receptor 1). NPR-1 explains differences in the worms alcohol sensitivity.
Researchers used alcohol concentrations comparable to those that causes intoxication in humans.
-Hawaiian worms showed a dramatic recovery from initial intoxication than did the English worms.
In accordance to the genetic analysis of the two strains led to the difference between them due to the
gene for NPR 1. NPR 1 act as a neuronal receptor-a molecular “socket” in brain cells.
Genetic difference caused the production of higher levels of NPR 1 protein in the alcohol-sensitive
English worms decreased their ability to recover.
Above figure depicts-
Former figure represents the England strain of worm with slower recovery and the latter one
represents the Hawaiian strain of worm with faster recovery.
Locomotion-
This figure shows one wild type (in upper left corner) and 5 locomotion defect worms. As it can be seen,
the moving pattern of mutants is different from wild type. This is to show how locomotion defect can
affect worm’s motion.
Natural Variation in NPR-1 gene modifies Ethanol Responses of wild strains of C. elegans-
Allelic variation that alters the functional level of NPR-1, a neuropeptide Y (NPY) receptor like protein,
can account for natural variation in the acute response to ethanol in wild strains of C. elegans
In C. elegans, including NPR-1 most of the proteins has been postulated to play a role in mediating
ethanol responses. Same tissue concentrations of ethanol that produce intoxication in humans have
neurodepressive effects on multiple behaviors of C.elegans. Level of response to ethanol-acute
adaptations that occur shortly after ethanol administration.
The NPR-1 gene encodes a predicted G-protein coupled receptor in the neuropeptide Y (NPY) receptor
family. Variation in the acute response to ethanol between genetic basis for differences in
human populations that individuals has a significant impact on determining have different
ethanol sensitivity and susceptibility to alcoholism.
Here we show that allelic variation that alters the func- variation in ethanol responsiveness have
yet to be retional level of NPR-1, a neuropeptide Y (NPY) receptor-. like protein, can account for
natural variation in the C. elegans is an ideal organism in which to analyze
acute response to ethanol in wild strains of Caeno- natural variations in behavioral responses
due to the C. elegans.
NPR-1 negatively regulates the powerful genetic tools available and the existence of
development of acute tolerance to ethanol, a neuroadaptive- multiple wild isolates obtained from
different process that compensates for effects of ethanol. Furthermore, dynamic changes in the
NPR-1 pathway neuronal proteins have orthologs in C. elegans.
Downregulation of NPR -1
Interestingly, clumping associated with ethanol exposure is only observed when removed from ethanol
Activating effect of ethanol removed from physiological environment leads to downregulation of NPR 1
pathway exposed. Animal with reduced or no NPR-1 pathway function demonstrate faster rate of acute
tolerance development .
Ethanol Exposure-
Behavioral Responses of C. elegans to Ethanol in the graph is shown below.
- Ethanol (200 mM) causes a small decrease in the amplitude of body bends.
- Complete flattening of the body bends can occur at 400 and 500 mM ethanol.
Characterized by their large conductance for potassium ion (K⁺) through cell membrane (leakage
channel). These channels are activated or opened when-
Opening of BK channel allow K⁺ to flow passively through the channel,down the electrochemical
gradient.
Efflux of K⁺ from the cell, leads to cell membrane ( hyperpolarization ) an increase in the electrical
potential across the cell membrane.
-neuronal excitability
BK CHANNEL-tetramer
BK channel is a tetrameric structure.
Each monomer of the channel forming α–subunit product of KCNMA1 gene, modulatory β–subunits
( KCNMB1, KCNMB2, KCNMB3 or KCNMB4 ) can associate with tetrameric channel .
STRUCTURE OF BK
A unique transmembrane domain ( SO ) that precedes the 6 transmembrane domains (S1-S6).
CTD contains four primary binding sites for Calcium (Ca⁺⁺) called “Calcium Bowls”.
C.elegans : Identification of BK as the major mediator of depressive behavioral
effects of Ethanol
Its approach for identification is “Forward genetic screen”.
BK channel plays a role in the development of acute functional tolerance (AFT) to ethanol. Loss of
functioning of SLO-1 results in inability in developing AFT.
TAG Lipase regulates to develop AFT. Manipulation occurs in level of TAGS( TAG lipase, lips-7 ).
By mutation : TAGS changes the basal behavior of phenotypes results to Hyperactivation of SLO-1 gene
development of AFT. This result provides genetic evidence for a link between lipid environment and BK
channel.
How long has NASA been studying C. elegans?
The first experiment utilizing C. elegans in space was conducted on STS (Space Transportation System)-
42, which launched in January 1992. The experiment was designed to study the biological effect of
exposure to cosmic rays. This mission contributed to helping scientists understand how to protect space
travelers on long missions.
It underlies my strong conviction that the rigorous, detailed and analytic study of the biology of any
organism is likely to lead to findings of importance in the understanding of other organisms.”
Cheap to maintain
Present no biohazard
-Easy to view Specific cells, cellular architecture which can be examined like GFP
- Individual cell lineages can be easily traced and directly inspected by DIC light microscopy.
Specific cells, cellular architecture can be directly examined using reporters like Green fluorescent
protein(GFP)* .
Ilys genes(lysozyme) exhibit distinct patterns of pharyngeal expression.
GFP= GREEN FLUORESCENT PROTEIN .Small protein derived from luminous jelly fish and can be
introduced as a transgenic tag to mark any protein in the animal.
2. Neurobiology
be conducted.
3. Aging
• Short life span helps to conduct genetic screens to find longevity genes.
The present study seeks to elucidate the mechanism by which chromatin organization impacts
organismal aging by studying C. elegans protein (Defective proventriculus) Dve-1.
• Generate C. elegans strains mutated in Dve1 and characterize the function of Dve1 during C.
elegans development and aging.
• Depression, Psychosis
• Parkinson’s, Alzheimer’s
• Pain .
Metabolic
• Type II diabetes
• Obesity .
Other
• Cardiovascular (arrhythmia)
• Oncology
• Muscle disease .
C. elegans in the Drug Development Line
Hence, Caenorhabditis elegans is considered as the model organism with enormous biological
applications.
REFERENCES
response
Classification
Introduction
C.elegans as a model organism
Similarities between C.elegans and Human
Comparison between 2 weeks organism and 70 years organism
Microanatomy
Reproduction and development
Life cycle
Drunken worm reveals a genetic basis of alcohol response
Natural variation in NPR-1 gene
Downregulation of NPR-1 pathway
Ethanol exposure
BK Channels and its Structure
Genetic evidence between BK channel and Lipid
Advantages of C.elegans as a research tool
Application in biological research areas
References