the making of the avian feather – an extreme lightweigth ...the making of the avian feather – an...

The Making of the Avian Feather – an Extreme Lightweigth Structure

1

��Chen Siang “Gene” Ng

Institute of Molecular and Cellular Biology&

Department of Life ScienceNational Tsing Hua UniversityNational Tsing Hua UniversityNational Tsing Hua University

1

Institute of Molecular and Cellular Biology

The Bird is a Special Kind of Reptiles

2

Feather Evolution

Sues HD. Nature. 2001 Apr 26;410(6832):1036-7. 3

Feather diversity

4Chen CF, et al. Annu Rev Anim Biosci. 2015;3:169-95.

5

Feather diversity

Chen CF, et al. Annu Rev Anim Biosci. 2015;3:169-95.

The flight feather is composed of the feather shaft (rachis and calamus) and the feather vane (barbs and barbules)

Materials Today Volume 20, Number 7 September 2017

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Physical (structural) colors of feather

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Materials property chart for biological materials: toughness versus Young’s modulus

Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015

Samurai armor

Keratinized skin appendages

9

Alibardi L, Toni M. Prog Histochem Cytochem. 2008;43(1):1-69.

Lee CH, Coulombe PA. J Cell Biol. 2009 Aug 10;186(3):409-21.

!- and "-keratins ��

10

Intermediate filament structure of ��keratin


Detailed structure of: (a) molecular unit of α intermediate filament: the heterodimer


Structure of the beta-keratin filaments


Schematic illustration of the biosynthesis of keratin


Schematic illustrating the development from an immature feather (pin feather) to an adult feather with electron micrographs showing the keratin development


Structures of body contour feathers and wing flight feathers at different growth phases

Ng CS*, Wu P*, et al. Genome Biol Evol. 2014 Aug 24;6(9):2258-73.

��Prof. Wen

�Prof. WenProf. Wen-

��Prof. WenProf. Wen-HsiungHsiungHsiung Li

��Prof. Cheng

�Prof. ChengProf. Cheng-

��Prof. ChengProf. Cheng-Ming

�Ming Ming ChuongLiLi

Diagram of normal developing and mature embryonic and adult chicken feathers

19Ng CS*, Wu P*, et al. PLOS Genet. 2012 Jul;8(7):e1002748

The frizzle chicken phenotype

Ng CS*, Wu P*, et al. PLOS Genet. 2012 Jul;8(7):e1002748

��Dr. Wen-Tau Juan

Sequence analyses identified a 23 a.a. deletion in a conserved region of KRT75

•The entire part of link L2 was removed.•Disrupt the structure over the coiled-coil segments of 2A and 2B


Contrast between feathers that have misexpressed GFP (Control, right wing) and KRT75-MT (left wing)


Striking differences in feather patterns on different body regions

23

Chuong CM, et al. Curr Opin Genet Dev. 2000 Aug;10(4):449-56.

Prum RO. J Exp Zool. 1999 Dec 15;285(4):291-306.

Prum RO. Nature. 2010 Nov 4;468(7320):E1

Central Dogma of Molecular Biology

RNA was extracted from epidermis of developing feather follicles after plucking

RNA Extraction!"

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We studied α- and β-keratin gene expression patterns in five types of feather epidermis

cEB

cLB

cEF

cMFcLF

34 α-keratin

genes

149 β-keratin genes

26Ng CS*, Wu P*, et al. Genome Biol Evol. 2014 Aug 24;6(9):2258-73.

27

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Ng CS*, Wu P*, et al. PLoS Genet. 2012 Jul;8(7):e1002748

The Structure of Feather

Structures of avian skin appendages and RNA-seq analysis

28Wu P*, Ng CS*, et al. Proc Natl Acad Sci U S A. 2015 Dec 8;112(49):E6770-9.

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Combinations of α- and β-keratin genes contribute to the morphological and structural diversity of different avian skin appendages


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Molecular Approaches for Functional Studies

Dominant Negative Mutation

Antisense

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Functional analysis using mutant chicken keratin forms based on those found in the human !-keratin mutation database led to abnormal phenotypes

Ng CS*, Wu P*, et al. Genome Biol Evol. 2014 Aug 24;6(9):2258-73. !"

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We utilized a retrovirus transgenic system to ectopically express mutant α-or antisense β-keratin forms

α- and β- keratins are not just structure proteins and also participate in gene regulation.


Combinations of α- and β-keratin genes contribute to the morphological and structural diversity of different avian skin appendages


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��National Synchrotron Radiation Research Center

��Dr. Yao-Chang Lee

Fourier Transform Infrared Spectroscopy

Internally reflected infrared beam in ATR crystal

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Feather samples

FOV

FIV

CNPP

plumulaceous

IR spectrum of chicken flight feather and counter feather

Spectral curve-fitting in IR spectroscopy for determining the secondary structure of protein in chicken FIV

beta-turns 1616-1618 cm-1

parallel beta-strand

1633-1635 cm-1

unordered structure

1643-1647 cm-1

alpha-helix 1651-1655 cm-1

beta-turns 1666-1669 cm-1

parallel beta-strand

1681-1683 cm-1

anti-parallel beta-strand

1690-1694 cm-1

FIV FOV CNPP plumulaceous0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

0.20

beta-turns parallel beta-strand unordered structure alpha-helix beta-turns parallel beta-strand anti-parallel beta-strand

FIV FOV CNPP plumulaceousbeta-turns 3.795% 7.27% 3.51% 4.55%

parallel beta-strand 18.43% 18.58% 13.52% 13.59%

unordered structure 9.02% 0.53% 10.31% 1.92%

alpha-helix 5.46% 14.58% 4.14% 13.65%

beta-turns 7.42% 6.73% 2.95% 11.17%

parallel beta-strand 3.95% 2.06% 1.63% 5.16%

anti-parallel beta-strand

2.83% 3.43% 2.80% 2.73%

β-keratin genes of Gallus gallus

Ng CS*, Wu P*, et al. Genome Biol Evol. 2014 Aug 24;6(9):2258-73.

Dynamic evolution of β-keratins in the archosaur lineage

Greenwold MJ, et al. BMC Evol Biol. 2014 Dec 12;14:249. doi: 10.1186/s12862-014-0249-1.

How can different morphotypes of feather be generated?

43

Fractal branching network

Blood vessel Lung

Feather Evolution

Sues HD. Nature. 2001 Apr 26;410(6832):1036-7. 45

We compared the gene expression patterns in different types of feathers and different portions of a feather and identified morphotype-

specific gene expression patterns

Ng CS, et al. BMC Genomics. 2015 Oct 6;16(1):75646

Tube development

Tube development

Vasculature development

Blood vessel development

The genome size, numbers of α- and β-keratin genes, and keratinized skin appendages of amniotes

47Ng CS, Li WH. Genome Biol Evol. 2018 Oct 1;10(10):2572-2586.

How can the diversity of feather be generated in the same species of bird?

48

Birds can be briefly classified into altricialand precocial species

49

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檔ப٢Chih-Kuan Chen

Regulatory Differences in Natal Down Development between Altricial Zebra Finch and Precocial Chicken

50

Chicken � Zebra finch ��

Chen CK, Ng CS, et al. Mol Biol Evo. 2016 Aug;33(8):2030-43.

51Chen CK, Ng CS, et al. Mol Biol Evo. 2016 Aug;33(8):2030-43.

FGF16 overexpression suppressed the natal down growth, reduced the bone length, and increased the epithelial thickness in E12 chicken

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Bimetallic stripThe frizzle feather rachis an asymmetric beta-keratin stripe

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Keratin nano-fibersunder X-raymicroscope

Surface profile ofthe interlocked

vane

! Prof. Cheng-Ming Chuong (?�)

! Prof. Ping Wu (��)

! Prof. Randall B. Widelitz

! Prof. Margaret Dah-Tsyr Chang (-�2)

! Prof. Chia-Wei Li (�'5)

! Prof. Chih-Feng Chen (.�()

! Prof. Hsu-Chen Cheng (9��)

! Prof. Ping-Chi Tang (&�,)

! Prof. Shuen-Ei Chen (."�)

! Dr. Yi-Chiao Chan (4�;)

! Dr. Wen-Tau Juan (��3)

AcknowledgementsPIs and Postdocs

! Prof. Wen-Hsiung Li (��1)

! Prof. Jason Isheng Tsai (8�*)

! Prof. Chih-Ming Hung (!�7)

! Dr. Mei-Yeh Jade Lu (�#:)

! Dr. Tzi-Yuan Wang (��)

! Dr. Wen-Lang Fan ($�%)

! Dr. Chien-Hao Su (B�6)

! Dr. Meng-Shin Shiao (<��)

! Dr. Meng-Ru Ho ( ��)

! Dr. Woei-Fuh Wang (�C@)

! Dr. Joyraj Bhattacharjee ()�))

53

Students and RAs! Chih-Kuan Chen (.��)

! Siao-Man Wu (�D+)

! Jiun-Jie Chen (.�/)

! Yu-Ting Lai (= �)

! Wen-Sui Lo (A�>)

! Jinn-Jy Lin (�0�)

54

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the making of the avian feather – an extreme lightweigth ...the making of the avian feather – an...

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