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REPORT DOCUMENTATION PAGE SRL
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TH RBSTRAET
‘hope elo thine (Usury tens of ala popes Doles He sha he abeieu 2 mgrech est
fr Fer hivemen, fe ade seid wih se gms Cangaced vith una enea so, he et
‘eeeplthave nce ajar! (Ueto df pesca fe reer apa achigh anit
{ernere i th serie nage ving mst ns are wrk ulead en ave oes. tbe eae of De
Ppresdeccuch nate tuolet a anand the lie ene, sel itn. and perrmance Melo TR
Fert, Tice eh fra wun recepls it ptepone uae Io Bala of he eae Wa 1 ae
evep aid inporea esis wssled ik sac and hc ap opie sea rooesng alge whch re aed the alld
Fectanum of tila! mse which es ipl iy of ves of xohdon famlvets clos: mapas,
[etas orl swollen a ao pps avestenr ine berm arora. singe aed rect sprouse oF Be
homes eer Rd hen impwowed Py cong sprain said absorbs wl ete eases, AaAHona at
tench nes mifete denote ne para rarer whch ne me pe sh Te
TE SUBTECT TERNS
uti
BEST AVAILABLE COPY
Final Performance Report
atroduetory Remarks:
‘The project focussd on the infarcd (28) seasory systems of so-called pyraphilous bests
‘shish show she beliavionr to approach forest fires. Fur this reason the beetles are equipped
‘with TR sensory angens. Cempered with uncooted tecuieal TR sensors, dhe insect IR receptors
have some major advantage: (i) here fs wn need for tomperatue conmsamey (Ihe reueptoes
‘optate at high ambient tempratuces; (Gi) de receptors ave rugged having a smart desian, and
work under horch environmental conditions, Additionally, the neuroual neon whi
processes the TR informacion in the cental nervous system ofthe beele, obviously cen
‘exracltherelavant signals fom considerable noise
‘The objective of the pruposea research ws f broadca ot unerlanding of the biolngical
Significance, speci lunction anu performance of biological IR reception. This included the
‘search for bitherto unknown Lt roceptors ia pyrophilous insect, The nal aim of the research
was wo farther develop unl impzave a technjal uncooled IR sensor and the apprnpriake signal
processing algevthuns which ae based wn the principles ane mechanisms ofits biological
_modola thigh were improved by millins of years of evolution,
To achieve ihe objectives, mumphological, behavioural, newophysiologial, asd therma-
physical investigatims have baen performed, Using the obtained results, the prototype oF te
[hotomerhant IR detector had been improved by combining appropriate sold absorbers with
sensitive mechanascnsors. &dditionaly. an approach ws made to detsomine some kmporart
Parameters which ure use ln spevify echnical IR seasons.
De, Helmut Sehmiv : PE
“Vice graduate swdents fished thelr Uipluma Ds under this grant,
1. The prothoravie influred organs of pyrophitons beetle Acanthocne
nigricans (Acamhoenemidac)
uring the fem ofthe grant a hitherto urknoom TR zeeeptor wae discovered in che Avstatian
“lide-ssh* boctle decamthocucenas nigra
(funily Acamhecnetidae}
Acanthoenenmus nigricans fs a small dg: becils having a body length of only 3-6 mun. The
species nigricans isthe only recent speview within the genus 4eanphoenemus and is distributed
allover Aust ts eontrast fo its incunspienous appearance, A. nignvates shiws
rernakable behuviour us beetles af both sexes are atacted by forse firs, The beetles imasle
fred
x bumt area immediately after the flames are extinguished and approach areas where
lowing remnants nf ees ce hot uses ure stil preseul, The reaany for this so-called
Pyrophilous belusiour i hardly understood bur it oan be syeculucd thatthe desetbed “pat
spots” serve as meeting places For the sexes. Must probably te Females deposit their eye
‘nto the ash or nade te burk of tom tees. Currently the lrvul food is unknown, Beaune
Jurwe wore obviously exported vu of Anstsfia inside wnod, its very probable thar the
corbin layer of feesly hurt res represents the primary source of food for the firs instars
‘Nothing was huavin about host receptacs in. nigricans. However pat of unusual
Ihypomeral stmemis fa font oF the fore eoxac has been deserided (Lanenence mud Bitton
1994), Bach structure consists ofa uy cunicular die (ameter 100-1514 ya) which could
serve as an absorber fur slectomagneiceadiston, We subjected these structures ta inept
ncspholoyical and physiological invemigaions with respect ob a possible fre-televant
sousory function. Ry out morpuclogival and wltastuctorsl investigations, we wens able co
Geseribe @ new type of inset sensillum which most protubly is involved in IR reception.
Material und methods
Azimol
Adull bates were colloted in 2002 and 2043 on freshly hum areas fa Westcen Asta,
-snimals were kept fr several was
in pantie boxes and fe ith sins, pean and
‘alate; drinking water was given ad ibid,
‘Scamsing leetron microscope
|2 utes were air-eed aad plved on hulders by LefeTahs (Plann, dinmete> 12 mum) with
ther the doysel or the Jtcral side. Sensory dive fm 7 beetles were isolated aud cfeaned by
sonication im argiatur of ehlevoform / ethanol (2:1) foc 2 min, Dises wore aired again and
tmoented om hders in an upright position which ullawed examination frm al sides,
Spoeimens were sputtered wilh gold and cxamined witha Cambridge Stereoscan 200 STM.
Lig end sranomission oleerton microscuny
Sensory dises af 19 heeles wore isolated al immodiately ieee in ive glutaraldehyde
fixative (9% glutaraldehyde in 0.05 mol I" cacodylate buffer, pld 7.1; osmolarity 386-400)
‘mosmol 7) and fixed overaipbl. The dlises were ther. washed in buifer pastfined with 1.5%
(2404 in the same buffer, delydisted hough an ascending ethuenl series and euibedded in
[Epon 812. Senvchin and ultrathin sections were cu witha Reichert ltracoe Mictototwe wine
lass or diamond knives. Seroichn seotions (0-5 unt} wore staid with w 8% tahun
blucthorsx sohiton sud exureined with 2 Leitz DM RDF light microscope, Digit images of
the sections were ikem with a Nikom Coolpix S000, Uleathinsestions were stained with
‘way acetal and feud vitro andl examined With» Zeiss EM L09 TEM,
BDerecunsiraction of the sensory dsc
‘Toinvecigate the nusaber el postion a sensory cell sencata inside the sensory dive, «sero
F210 semithin erose-aserions was eul Uusugh a sensory dise. The digital fmages of the
seoions wes
stored um a PC nnd the swage Asaira (Version 3.0, TG Toe, San THego.
USA) was ued for 2D recoustruction. Dus te inferi quality, 34 om ofthe 240 images could
snot be used. Therefore, the distances bern tn images were adjusted indi vial to
‘eampensite forthe Lose of thse seston. Tis outer cell merubraues of te sormu could mot
be rave and identified reliably, There, we used the mele and ma
coli of the sensory
cls for soma idewitication. To ewensnty setomatic dewtfiction bythe wnftvare, oncel aud
‘nucleoli as well asthe cuticle were cotoured in cach section usiug a digitizing tay
Riscooplnsiology
exracellarelectrophsiological eocardings were performed fn prothoracie discs from 4
beelles by clesroltically shaspened tun
buticle of the dise by a miceomanipulator. To minimiza daraays ofthe dive bythe eecteode,
lecenodes hich were gradually driven int the
pence a ae culicte was stopped a5 soon as neneenu] uclvity of the apsmuunevisly active
units was envonutered, Signals were amplified 10 x and displayed on an oscllosoape, In
onger to mie the natural ston Gj a forest Bre) we stated the IR organ sith smell
‘orm emiller which wus mounted 40a assay Conn the beetle an onl he ate ty 400°
CC. The heede was glued with its eu! side up ome a plate und w shuller beeen the hot
emitter and the beetle allowed defined cxpasure to ce die chermal radiation. The to
dimersiqnal surise af the emitter, uel towards Une beetle, wast? x 25 cm. fn a serien of
‘oxpevingnts we irae the peufhoran wil Une sensory die for J soe. fo etimste The
ceortespondixg increase in owicula temperate, we used any thermocouple (Fype K:
‘Chromega’Alomega. diameter of the bead 33 wen) Hecause it was technically impossible to
mou the bead undor dhe outcr cuticle of the intact sensory dite, we glued it with « mint
amopnt of Syndeticon ln the inaer surface af upfece of abdaminal cuticle of su ar-ried
bere. As the abdominal curcle (abou 15 ym} is Twice as thick as cutie from the cuter side
ofthe sensory dise 7 $ umm), temperarae wales mia be slightly underestimated, The wires
ofthe thorm:cenuple were connected ta an electronie thermomcter (HHL 202. Newport Omega,
aveurary +9:2°C) and the cuticle wus expose tn the thermal ominer from distance of 40
sum, Mean texmperarate increase of the entice was 4.7” C = 0.78 € (n= 8 expen
Results
‘The protiorate TR organs
‘One paie of TR organs is Toeated on the ptothoems in bth sexes. The io organs restated
itctly enterin tothe coxa of the protharacic Tepe (Fig, 1), The most suing eamponemt of
an IR urgsa isa ure oe less tound cuticular dis hich is sumznc sumken inf the surfiee
of the prothorm. At its pasar edgs, the dite is held bya stalk. hich arigiates a the dons
anmerior border ofthe coxal savy. Below the dive, a hemisphsrial air-filled eaviy is
situated. Because the opening of te caviy is somewhat larger than the Uiemeter ofthe dive,
‘xchange of air hetvtcon the ouvity andthe outside ts possible, The die, which has beewe
Found tn be ionervated by sensory eal somata (Sclunit/ etal, 2002) represent the seta TR
roeeptor, whoreas the eaviy esr he rogarded as an auxilinuy siucture
Outer morphology of he sano die
A sensory dis has a dlamoter helseen 10 uni 18S pam, The size roughly oceelates with the
shat ofthe beetle, In some beetles, she outer surface appears slightly carved ina wherews
the surface af the underside always is bulged out conexty (Pigs. 28-D).'Ihe stalk shows an
‘ovat esos sation witha nae diameter of abe 30 ym. We Found no indication tha the
stall has any exible properties.
‘Namerous small sexslfa can be found om the ayter surface of he dic Fig. 2B}, Hawever he
seruilla ure not evenly dist ted. Most seuss concentrate with an anterior sericiceular
area an the npper surface. Some senile (abut 13 %,N +9 dises) were even found on the
anterior edge ofthe dae (Fig. 2€). We nover found a sensillum am the underae Mast
‘rububly, all seuitla baloee to the seme type and san be identified by their cuticular
anparulus which consists of w seal cullule peg susvounded by a socket, At Sis bas, the pex
has adiameter af whoul2 wom anda soa! rapt of about 35 a.
Fig.2
Morphology end wlrtracture of he sensi
Series of cross and longiradial soosions through che sensitin evento vat the sma wulioular
peg vonssts af massive cuticle. Nether pores nora Tnmen inside the peg was fount, roan
below, the peg is innervated by a single itary receplor cell (se reconstruction ofa sensilla
{iv Fig, 3), The dendrite onter segment (1005) i eylineeelly and anbranchel. The DOS has w
‘diameter of ess Ulan 0.5 psu and contains oaly sparse neunweubol. Ava spoeifc festore, the
entire DOS in emsbeatbed by an electron des aateriel whieh most probably i he
hypertphiod dendrite sheath, The oulermunt tip of Ibe DOS terninutes whe base uf the pe
ad is comnocied te its cutee byt presumed dendritic sheath whioh — atleast fn some
sttlons seems 1 insert ito tke basal pat of the peg, No mibular body was found,
A ciliary sonsaietion subdivides the dendritic outs (DOS) and the dendritic inner segment
{DIS), However, becanse dhe DOS shows already a small diameter, thae is na decreare in
iumeter (hum the beso! part Uke DOS lawanls the dendiic coastiction. Two basal bodies
‘were fond jut below the eonsircton. The husal dies were intereumectel by cool
‘laments which donot extend nv furiser int the proximal region of ths JIS. Below the
ccliury constietion the DIS strongly broadens. As a peedominnnt festa, many deep
invaginations ofthe cell memirane into the fumen ofthe cell wete abserved. Thin elke
Processes of glial cell ure squeezed besseen the menabranes aad a lage nunc of
smuwehoudtia weze ays found inside the DIS.
Ilse same region eval he easily idealfed by 4 lage muclens hut the BAS and the soma are
‘at clealy dstingnishable as thy slinw the seme sractural features (Le. thin but deep
Invaginations of glial cells nto the call Juen and tots of mitochondria), We thund 2-8
the thecayen, wichogen and lutmagen cel)
eaveloping cells vost probably represe
‘which aminly enwrop ne dendsie aud the distal region af ta soma coll, Consequently, mest
‘of the oall soma is cnvefopet hy a thin lil Inver. Tn general, 2-3 nuclei of enveloping cells
‘were found in the region ol the eiliaey coustistion, Die tothe trom entanglement af ell
‘material inside tho disc, an unequivncal identification of snwle enveloping ells could ao be
made, Based on commplatessucs of wrath sections Uyrough atoral ofS sensilla. graphical
sconnstructinn ofa singe sensilhum was mate (Tig. 3.
Fig.3
First eectrophysiologleed recordings from the multipolar new ones ince the disc
‘The recordings toma the sonsony lise (N'= 4 hestles) she that Oke mullipolr news had
omgarng activity al toms emaperatare, The thequeny’of spomsanones activity of single units
was hetwcen § and 1m Hand eas unaffected by sound (Goice or hand clappiug), moderate ait
sruvemuents or goals toneh of the sutras
ice hy a bristle. To contcas, exilim 12
he VK ration from Ghe Use emit iacteaced fing ftequeney in aphasie-tonic way
(ig, 4), Wo also stimotacd the receptor with eur wr ol sie ight Com a eedtiebt
Delium-neom lave, which was used In former swine to simple the 1 organs of pl vipers
dependently of how the temperature was increased, an increased spi ftequeney reli
In geuetal. a higher steedy-satevompersture was ceded hy u higher discheeve Lequency of
‘he units. Censation of dhe sinus dahibited the generation of aston potentials
SUMMARY. 4 migricunc has one prof wigue profborace sensory organs which most
probably serve a infin reespmns, Hach organ enasists of a euicutar dsc which is xe
‘over au nielled cavity, On the aur settee ofthe die, aber 10 tiny euticular sensilla are
situated, The poreless ter peg ofa sensillum is 3.5 pon Jong and is suirounded by &
cuticular wall One ciliary sontory cell inmervates the peg, the dendle of itch I divided
nu) an euler and et inner dente seamoat, A tubular body could nat be found inthe
‘outer dendritic tip. As a special feature, the outer dou
slzcttondenge marc, which moat pecbshly represen the hypertrophied dendeitic sheath
segment is omvcloped by an
“The Lunar dandsitie pment and the soma ars feted indistinguishably farming a commen.
ccTlulur space. Tin, leulike exeusios of ela] ells deeply extond inte thet onlarged laren
‘which glen contains large numbers of milochondein, The sill othe sensory dst of A.
lgptoans Sbsionsls sepeoscat a new type of inset sensilum of hitherto unknuven function
Tint electophyslological investigations indleate thet hey may funetion as wer ossplors
Publication
TL. Sehaity, A. Schmliz 8, Trenner,H, lockmann (200%) A new type of iecl inured
congan of fow thermal mass, Naiuneistensthufen 89: 225.209
E, Rrolss, A. Schmitz and IE. Schone: Moephology oft specialized sensitfa iu the
flothoracie sensory organ of Acarwhoensms nigricans (Clooplena, Avanthocnensiaae)
Arthroprad Structure and Developavent (subuaited for pbLin Artleoped Soreeare and
revetopmerd
sls thesis: Diploma Uuesis uf punduate student Bva Kecis (finished ia Avgost 2003)
tte: Punktionsmenphologische Churtersieruny der seciulsierten Shaneszallen des
prothorakalen Intrrotorgans von Aearthacenw nigricans (Coleoplea, Avuntaocaetilae
iptornhelt, 2003, Fostilu fir Zale, Universit Bon}
Tl Physiology and evolution of the inftarcd receptors of pyrophilous beetles
‘the genus Merinma
A therrmumenstive multipolar neuron innervates each of the four abdominal IR receptors of
ho Avstrafian hupeestid boctle Merinia arena. Flectrphysinlogicl mvordinus showed Ut
ie neuron is ative niin g oad snngs of body temporsins (ested bemtcan 10° Cand 40
(). We heated ie caceptors wit a ced diode laser
5 men? up © 1. Wiens In general, warming caused an inerense of receptor activity.
sued 100 9 300 ms aller onset of itadiation. Aor peak
66 ue) at intusitics ranging From
Penk discharge frequencies were
ficqnencics were reached, ditinet adptatin tnok place wilhin secs (Fig, 5). A linear
Jnerease in itmdiation intensity eansed an exponential! incase in peak frequendisy. Lawes
‘hreshold wane fund fo be ol 40 mes? where latencies were 47 ms, At the highest imensity
‘ested (1.3 Wren), pon Froquoncics increase up to about 300 Hz und lulencies Tecrased to
2am
