!"#"$%&

!"#$#$%&"'()*+#,#$"&-&./0)12234(5',2)+(--(4.) 6#-7("&)$%&).'8&)9):;,<=#,)#:)$%&).&>;(445) ?'-#"7%'<).7;".)9)<";"(4)24(,?.)',)$%&)-#,#$"&-&.@) ?*=2I63@&N=*0&@634& 5A)42B'0=C0D&>1A(D7=E&& FA4)38A)32&G3@24=*(=& & ;31CBH(**0D&?*=2I63@E&& ;=4C04&408()7&)7&J=6&

V4=72&R&F0J6*0B5J(2A.&$WW#&& ?*=2I63@&O0J=*0&@634& ?*=2I63@&N=*0&

>1A(D7=&N=*0&

K&-&L00C@M& P)A7@2)7.&N=DD07.& SA(T782)7&R& '()*+&,$-.&/012340&$#.&564(78&9:$%.&;4+&<=407&>+&?0204@07&& Q(1)*@)7.&R&?I70.&9::%& '0*)U.&9:$,&&

A%&,)?'?)$%'.)B&,#-)?&4'B&"5).5.$&-)&B#4B&C) !4($57;.)9)1<%'?,(0).&(.#,(4D)(..#<'($&?)E"&&?&".) Q(1)*.&G7D04@07.&R&P)70@.&9::!&& X434=*&8*=7D@&=40&D04(U0D&Y4)J&J)D(Z0D&=6)14(70&[6A04)J)70\&@L0=2&8*=7D@& N=*0&5A)42BH0=C0D&>1A(D7=E& H400D(78&a&,&I4@&

Ab6E""LLL+=4C(U0+)48"6*=2I63@" )47(2A)4AI71A3@B=7=c73@"U(D0)B:W=&&

?A04)J)70@E&D(]04&HI&@0^.&(7D(U+& _=44(@.&_)**=7D.&X=J04)7&.&02&=*+&9:$,& ;(=J)7D&`&(7D(U+&U=*30&

N=*0& ?*=2I63@&

SA(T782)7&R&'0*)U.&9:$,&& V4=72&R&F0J6*0B 5J(2A.&$WW#&&

$& !"#"$%&

A%&,)9)F%&"&)?#&.)-(=,2)#<<;")',)G(.-(,'(C))) K&.<"'E&)$%&);,;.;(4)"&7"#?;<=B&):&($;"&.)9) A%5).#)-(,5)-;4=3-(4&)*HIJ)#:)$#$(4/)2"#;7.C) E&%(B'#")*-(=,2)$"(',./)#:)$%&)-(4&)&<%'?,() N=c78&84)36&*)1=c)7@E& N)44)L.&G7D04@07.&R&Q(1)*.&9::W& 9%d&A)**)L&*)8@& >1A(D7=E&$&@(D0&4024=12@& $-d&H344)L@&R&D0640@@()7@&(7&@)(*& 64()4&2)&J=c78& $,d&37D04&4)1C@&)4&4)1CI&)U04A=78@& e2A04E&&U=4(0Df+& '37D*0@&[K$::&*=480&@604J\& Y)4J&(7&06(D(DIJ(@E&Y=@2& V4i2h704.&Q(^)7.&R&P)70@.&9::#+&&

N=c78& F4=(7@& Y4)J&$WW%&2)&9::#+&[7`!9\+&& ;(@24(H3c)7&)Y&J=c78&84)36@&

Ab6E""LLL+j)347=*@+31A(1=8)+0D3"D)("$:+$:#%"!99#,-&U(D0)& P)A7@2)7.&5J(2A.&?I70.&02&=*+&9::-) Ab6E""LLL+D=(*IJ=(*+1)+3C"70L@"=4c1*0B9--:gg%"'(h=440B01A(D7=BJ=c78B4(23=*B1=62340DB1=J04=BJ=*0@BY)**)L(78BY0J=*0B*(70BF>QBA)34@BD=I+A2J*&& Ab6E""0U)*3c)7)YJ=JJ=*@+H*)8@6)2+1)J"9:$!":%"2A0B0U)*3c)7B)YBJ=7D(H*0+A2J*&&

K&.<"'E&)$%&)"&7"#?;<=B&)$"(<$)#:):&-(4&)-#,#$"&-&.) K&.<"'E&)$%&)-&$(E#4'<)(<=B'$5)#:):&-(4&)1<%'?,(.) ?*=2I63@.&)7*I&*0k&)U=4I&D0U0*)60D+&& >1A(D7=&6)31A&D0U0*)6@&=2& @2=42&)Y&J=c78&@0=@)7+&& >^2047=*&20J604=2340&D=2=& 401)4D0D&Y4)J&$&Y0J=*0+&

•! ?40@3J0D&Y04c*(h=c)7&!&$& •! O(7=*&=4)3@=*&!&9& •! >7204@&734@04I&!&g&&

_(H047=c)7&

V4i2h704.&Q(^)7.&R&P)70@.&9::#& N)44)L&R&Q(1)*.&9::W&&

9& !"#"$%&

A%($)'.)$%&):;,<=#,)#:);$&"',&).&<"&=#,.)',)-#,#$"&-&.C) A%($)("&)$%&)<%(44&,2&.)#:)&>$"&-&45)(4$"'<'(4)%($<%4',2.C)

N=*0&FE&)607& o713H=c)7E&088&(7&6)31A.&$:B$$&D=I@& O0J=*0&?4)80@204)70&`&X*)@0D& Q(1)*.&G7D04@07.& R&P)70@.&9::!+&& $9BA)34&mg:BD=I&)*D&6388*06388*0n&[>1A(D7=\&&(7&6)31A&

V0@2=c)7E&64()4&2)&088B*=I(78.&g&L00C@& 5IJH)*@E&D(]04072&6)63*=c)7@&)Y& •! 5J=**&088@.&gB,&JJ& 5A)42BH0=C0D&>1A(D7=& •! V=(7&U)*3J0&(7&32043@& p(@J(**04&R&N1<0*U0I.&9::g& •! 52=I&(7&32043@&37c*&$WB&2)&9:B@)J(20&@2=80&

Ab6E""LLL+=4C(U0+)48"@A)42BH0=C0DB01A(D7=" 50*L))D&R&P)A7@)7.&9::%l&Q(1)*&R&G7D04@07.&9::-& 2=1AI8*)@@3@B=13*0=23@"U(D0)B:W=+A2J*&&

A%($)'.)$%&):;,<=#,)#:)$%&)+L!)',)-#,#$"&-&)-'4MC) G#7'<.)$#)"&B'&F)9);,?&".$(,?) N/?&`&37(q30&J)7)240J0&*=12=c)7&64)20(7& 1.! Draw a phylogeny of these mammalian groups: Prototheria (monotremes), Metatheria (marsupials) & Eutheria (placentals). Place these traits on the phylogeny: lactation, sexually dimorphic venom gland & >=4*I&B&>1A(D7=& extra-tarsal spur, loss of venom function in the crural gland. /=20&B&>1A(D7=& 2.! Describe the adaptive function of the large male extra-tarsal spurs in the reproductive behavior of the /=20&?*=2I63@& N/?E&634(Z0D&N/?& platypus. Diagram the change in crural gland weight with seasons & explain the trend. How is the g!& function of the crural glands modified in echidnas? Trace the path of pheromonal signals in the brain, g:& as learned in quiz section. 9!& 3.! Describe the reason for the mating train in echidnas. The sperm bundles of monotremes travel 2-3 faster than typical mammalian sperm. What does this suggest about male-male competition? Describe 9:& the design of the penis of monotremes. How does it function? $!& 4.! Birds typically have 1 functional ovary. Compare the monotreme ovaries to other mammals & birds. $:& 5.! Diagram the cycle testosterone production by echidnas or platypus over a year. When is their breeding season? Why would the breeding season shift later & later in a north to south gradient in Australia? !& N/?E&634(Z0D&N/?& 6.! How did scientists use temperature recorders to track behavior of female echidnas? Compare the :& temperature patterns of a female hibernating vs. active above ground, vs. in a den. Predict the /(6(D@& ?4)20(7& X=4H)AID4=20& production of estradiol in females over an annual cycle & compare that with progesterone production by females. What are the hypothesized functions of estradiol & progesterone in females? V4(jZ2A@.&V4007.&/01C(0.&02&=*+&$W#,& 7.! Compare the gestation & incubation periods in the Echidna. What changes occur during gestation? R&V4i2h704.&Q(^)7.&R&P)70@.&9::#& Describe the size & yolk content of monotreme eggs. Describe the altricial hatchlings of monotremes. 8.! How does a female echidna take care of her young? Why is maternal size correlated with juvenile QFE&7)&240=2J072l&'5GE&H)U(70&@043J&=*H3J(7& mass at weaning? How does lactation differ early vs. late for the echidna? Hypothesize why the milk '=1(24=1(7E&H=1(24=1(7&=7cH()c1l&&F'5E&F4(@&@=*(70&H3]04& changes over time. What are the benefits of the unique Monotreme Lactation Protein? Ab6E""LLL+=4C(U0+)48"6*=2I63@")47(2A)4AI71A3@B=7=c73@"U(D0)B:WD&& >7j=6))4(.&V4=72.&Q(1)*.&02&=*+&9:$,&

g& !"#"$%&

Selected References

1.! Australian Platypus Conservancy. 2013. The platypus: A very special Australian. http://www.platypus.asn.au/ 2.! Enjapoori AK, Grant TR, Nicol SC, et al. 2014. Monotreme lactation protein is highly expressed in monotreme milk and provides antimicrobial protection. Genome Biol Evol. 6(10):2754-2773. 3.! Grant TR & Temple-Smith PD. 1998. Platypus biology: Recent advances and reviews. Phil Trans R Soc Lond B 353(1372):1081-1091. 4.! Grijfiths M, Green B, Leckie RMC, et al. 1984. Constituents of platypus and echidna milk, with particular reference to the fatty Acid complement of the triglycerides. Australian Journal of Biological Sciences 37(6) 323 – 330. 5.! Grützner F, Nixon B, & Jones RC. 2008. Reproductive biology in egg-laying mammals. Sex Dev 2:115–127. 6.! Harris RL, Holland BR, Cameron EZ, et al. 1,2014. Chemical signals in the echidna: differences between seasons, sexes, individuals and gland types. Journal of Zoology 293: 171–180. 7.! Johnston SD, Madden C, Nicolson V, & Pyne M. 2006. Identifying the sex of Short-beaked Echidnas. Aust Vet J. 84(1-2):63–65. 8.! Johnston SD, Smith B, Pyne M, et al. 2007. One-sided ejaculation of echidna sperm bundles. Am Nat. 170(6):E162-E164. 9.! Julien P, Brawand D, Soumillon M. 2012. Mechanisms and evolutionary patterns of mammalian and avian dosage compensation. PLoS Biol. 10(5):e1001328. 10.! Kuruppath S, Bisana S, Sharp JA, et al. 2012. Monotremes and marsupials: comparative models to better understand the function of milk. J Biosci. 37(4):581-588. 11.! Lefevre CM, Sharp JA, & Nicholas KR. 2010. Evolution of lactation: Ancient origin and extreme adaptations of the lactation system. Annu. Rev. Genomics Hum. Genet. 11:219–238. 12.! Liem KF, et al. 2001. Functional Anatomy of the Vertebrates: An Evolutionary Perspective Third ed. Thomson Brooks/Cole Publ. 13.! Livernois AM, Graves JAM, & Waters PD. 2012.The origin and evolution of vertebrate sex chromosomes and dosage compensation. Heredity (Edinb). 108(1):50–58. 14.! Menkhorst E, Nation A, Cui S, & Selwood L. 2009. Evolution of the shell coat and yolk in amniotes: a marsupial perspective. J Exp Zool. (Mol Dev Evol.) 312B(6):625–638. 15.! Morrow G, Andersen NA, & Nicol SC. 2009. Reproductive strategies of the short-beaked echidna – a review with new data from a long-term study on the Tasmanian subspecies (Tachyglossus aculeatus setosus). Australian J Zool. 57:275–282. 16.! Morrow G, & Nicol SC. 2009. Cool sex? Hibernation and reproduction overlap in the echidna. PLoS ONE 4(6):e6070. doi:10.1371/journal.pone.0006070 17.! Nixon B, Ecroyd HW, Dacheux J-L, & Jones RC. 2011. Monotremes provide a key to understanding the evolutionary significance of epididymal sperm maturation. J Andrology 32(6):665-671. 18.! Nicol S, Andersen NA. 2007. The life history of an egg-laying mammal, the echidna (Tachyglossus aculeatus). Ecoscience 14(3):275-285. 19.! Nicol S, Andersen NA, & Jones SM. 2005. Seasonal variations in reproductive hormones in free-ranging echidnas (Tachyglossus aculeatus): Interaction between reproduction and hibernation. Gen Compar Endocrinol. 144:204–210. 20.! Oates JE, Bradshaw FJ, Bradshaw SD, & Lonsdale RA. 2002. Sex identification and evidence of gonadal activity in the short-beaked echidna (Tachyglossus aculeatus) Monotremata: Tachyglossidae): non-invasive analysis of faecal sex steroids. Aust J Zool. 50:395-406. 21.! Oftedal OT. 2012. The evolution of milk secretion and its ancient origins. Animal 6(3):355-368. 22.! Rismiller PD, & McKelvey MW. 2003. Body mass, age and sexual maturity in short-beaked echidnas, Tachyglossus aculeatus. Compar Biochem Physiol A 136:851–865. 23.! Selwood L, & Johnson MH. 2006. Trophoblast and hypoblast in the monotreme, marsupial and eutherian mammal: evolution and origins. BioEssays 28:128–145 24.! Wagner GP, & Lynch V. 2005. Molecular evolution of evolutionary novelties: the vagina and uterus of therian mammals. J Exp Zool. (Mol Dev Evol) 304B:580–592 25.! Waters PD, Wallis MC, Marshall Graves JA. 2007. Mammalian sex - origin and evolution of the Y chromosome and SRY. Seminars Cell & Developmental Biol. 18:389–400. 26.! Whittington CM & Belov K. 2014. Tracing monotreme venom evolution in the genomics era. Toxins 6(4), 1260-1273. 27.! Williams GA, Serena M, & Grant TR. 2013. Age-related change in spurs and spur sheaths of the platypus (Ornithorhynchus anatinus). Aust Mammalogy 35(1):107-114.

,&