the making of the avian feather – an extreme lightweigth ...the making of the avian feather – an...
TRANSCRIPT
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
��
��
��
��
���
Physical (structural) colors of feather
!""#$%&&'(')*+,-'..'/01"/23)$-034&!056/23)$6+'7*6(0.0381.68*'"!*3$&
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
Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015
Intermediate filament structure of ��keratin
Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015
Detailed structure of: (a) molecular unit of α intermediate filament: the heterodimer
Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015
Structure of the beta-keratin filaments
Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015
Schematic illustration of the biosynthesis of keratin
Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015
Schematic illustrating the development from an immature feather (pin feather) to an adult feather with electron micrographs showing the keratin development
Wang B, Yang W, McKittrick J, Meyers MA. Progress in Materials Science 76 · July 2015
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
Ng CS*, Wu P*, et al. PLOS Genet. 2012 Jul;8(7):e100274821
Contrast between feathers that have misexpressed GFP (Control, right wing) and KRT75-MT (left wing)
Ng CS*, Wu P*, et al. PLOS Genet. 2012 Jul;8(7):e1002748
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!"
#$%&'%()%*+,%-*)$.(,%!//!%-01%!234!/567289:8/;<2!,
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
���
��
��
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.
�
�
�
�
Combinations of α- and β-keratin genes contribute to the morphological and structural diversity of different avian skin appendages
29Wu P*, Ng CS*, et al. Proc Natl Acad Sci U S A. 2015 Dec 8;112(49):E6770-9.
�
�
�
�
Molecular Approaches for Functional Studies
Dominant Negative Mutation
Antisense
30
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. !"
!"#$%&'()*+#+ ,-))(+.+#'")%/01������2304(5)#6789%.&. :*"%
;<=>? @ABA8CAB=?
D<EF8G<H<?
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.
32Wu P*, Ng CS*, et al. Proc Natl Acad Sci U S A. 2015 Dec 8;112(49):E6770-9.
Combinations of α- and β-keratin genes contribute to the morphological and structural diversity of different avian skin appendages
33Wu P*, Ng CS*, et al. Proc Natl Acad Sci U S A. 2015 Dec 8;112(49):E6770-9.
�
�
�
�
��������National Synchrotron Radiation Research Center
�����Dr. Yao-Chang Lee
Fourier Transform Infrared Spectroscopy
Internally reflected infrared beam in ATR crystal
���
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
�� ��
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
!""#µ$!!"#
!"#$%$&'#'()*$+#,
-./,
-.0,
µ$%
!"#$%&'( )(%(*+,&-+./ )+0+,&-+./
&
'
()*+,,-#./#0-12.3)3#4.52#*.66)0)/5#7./*3#86#39$$)509
Bimetallic stripThe frizzle feather rachis an asymmetric beta-keratin stripe
CMU��� AS������� NCHU���
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
㈖⹀劅猺⫛⧁孉䛟猻 壨≊傛猺㣊㤻䒀濆猻䙬㤒↢猺䇵疵蓦⎔猻
扽㤒⣪猺⪤痌翗階猻 ∶ㆎ猺⪤痌翗階猻
╪勝濕猺䫵䵗▊䛧猻 楔乒嗽猺榿办㌈䖡猻
㩸䲪峡猺办㺼妬䥞猻
崒哳䜽 㨯䚿㉅ 堂∲
55