Genomics of sensory systems - University Of Maryland

Genomics of sensory systems - University Of Maryland

Lecture 13 : Mechanoreception 10/14/09 Science / Nature papers

Broadly important phenomena Short format (4 pgs) Very dense

Multipanel figures Long figure legends Equivalent to several regular papers

Clear writing Questions 1.

What are the mechanoreceptive molecules? How do you know if you found the right molecule?

2. 3. Are the molecules common

through all organisms? How is the signal transduced? How do you prove a proteins function?

Find a mutant or mutate gene knock out Knock gene in to rescue mutant Show similarity to known sequences in other organisms (homologous)

Label and express - see if gene is turned on in right cells In situ expression of nompC in sensory organs

macrochaete microchaete

Proboscus bristles Chordotonal organs of halteres

Walker et al fig 7 Halteres also beat and are used as

gyroscopes during flight Chordotonal organs are used to sense beating

and provide feedback during flying Skin connected to

cytoskeleton by receptor Ion channel

Tubulins MEC - mechanosensory proteins identified from NOMPC

Textbook example - nompC NompC is new member of the TRP family of ion channels

GFP expression of nompC in C. elegans

Li et al 2006 : Sixth sense in worms Trp-4 = CeNOMPC How is it working in the worm? What are the function of the cells

where it is present? Li Figure 1

Li et al 2006 Trp-4 mutants move differently Bend more

Bend more frequently Putting trp-4 back in returns to

wild type Rescue TRP-4 expressed in the dopamine neurons

Dopamine neurons Interneurons

Wormbase Expression of TRP4 in dopamine neurons

Supp Fig S1a,c CEP neuron Li Figure 2 - What do these cells sense?

Li Figure 2 - What do these cells sense? Cat-2

DVA neuron - expresses trp-4 Expression of TRP4 in DVA and DVC

Supp Fig 1b Fig 3 Shimomura isolated fluorescent

protein from jellyfish Crystal structure Structure is made of barrel from 11

beta sheets and one alpha helix going up center S65 - Y66 - G67 react to form a

chromophore which fluoresces GFP naturally fluoresces

Absorbs light at UV or blue wavelengths Emits light in green

GFP can be used to monitor when and where a gene is expressed GFP expressed in mechanoreceptors of C. elegans

G-CaMP Ca+2 M13 M13 chain

of myosin light chain kinase cpEGFP

CaM Ca+2 binding protein GCaMP [Ca+2] dependent

fluorescence Inc in Ca+2 Nakai et al 2001

100 M Ca+2 Express GCaMP in DVA cell Tie down end of worm

Trp-4 mutants - no Ca+2 signal Li Fig 4

Are you convinced? Does trp-4 transduce bending motion? Is this the whole story?

What other experiments could they do?

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