BIOELtCTnIC RF.

AC':IOIiS or TIlt :UDBRAItl TECTUH OF

CHICK EMBRYOS ro LICl:T
ZhW'Dal EyollUtalonnoy lUokh1ll11 i H. L.
!"hiol0611. Vol., 1, lIo. 2, pp 176-1e2,
1965. Laboratory of Compa.rative Onto
of the Nervous System,
Brain Institute ot the
or Medical Dciences of the UGSR,
Molcow.
ABSTRACT. Ilectrieal relponsea to ,taulat1on with single,
rhythmical or paired flashes vere recorded t'r0lll the dorsal
lurtaee or the Illidbrain teet\llll. Potentials evoked by single
fluhe. were shown to occur in the contralateral midbrain
teet... .. the 13th dl or incubation. If. ::lont. of the
cues, relponsea ..lUlled a tn-phasic pattern (pas! t1ve-neRa
Uve-positive), the negathe eoaponent of larger UIJll1
tude. In emb!'Joa, U1P11tude ot the nep,atl "Ie CClIImOnent
vas tOWld to vary vithin to 00 \.IV, in 19-1lay Hlbryo!l - ",i tl:
in 50-100 being over 100 In 20-day Both amnl1
and shape ot the eYOked potentials considerable
individua1 variations, they alftO tended to verI in the StlllM! embryo
during an experiaent. The to rhythmical flashes shoved
180rhytbmicity at frequencies uJ) to 5 t'luhes per sec 1n
en'bJ7oa, and l1Jl to 10 nuhes per sec In 2o-da,y embryos. On
ulation \I1t.h pairod fluhea, the ablolute retractive period YQ
tound to lut 60 to 200 lUec In 18-2o-da,y eIlIbr:--oa. It haa thus
been shown,that in the chick eabr,ro tunctional develOJlllent or
tbe v':.ual analyzer attains a fairly high level in the last rre
natal
-1
the attention of' nhY3iolor;i3ts has bep.n ,lravn to a stu<!y
of the of the fUnctional maturetior. of different
development ot functions of the visual as one of the
main 1n terrestrial vertebratc5 has
:!etno;iG of evoited responses. w:lich arise ir. dU'ferent sections or
';hc r_"le.l:.::cr uron !l.:lequate atir.lulat1on ".:.1.(> i lle. "T nurinp. pl{'c
exci tati(;>n of tile optic nerve. ha.ve fI.1.wnys seemec1 pr01"klsinc for t:X
plar.ntior. of 1:E'culiarities or <levelo:nment vi SUFI.: fu.'1ct.ion in
cess 0: or,tofer.esis (1-10]. I1owe.,er. s'.1ch invc:>tigeticrlC have mostl:r beer.
limite.i to t:,c period of postnatal lire. ':'here have heen only at
tempts at. stucyi:"lf, ooteDt1als durin'; :'i,e ..or,ic Tlcriod C:cve:o""
Lent of f01Jl [11). paJor1ty of clcctronhyslo1op;icnl ',Ior':fi car
rleri out on vnrious ::J.a.umAls (rabbits. cats, ::1onkevs, etc.) nT.u.
presentutivcs CJ!' other classes of vertebrates. in "articulpr fowl. hav:
been in this respect.
t'c "'al!ted to trace the process of t:t<> functicnf\.l ma.turation of v<tr
1.ous secticr.s of the visual analyze.- In fowl. from the early ::t!1l;es
or entr/or.ic ti.eveloptlent. Thia work Is tl.e first sten in evoLed. r":!
eponses of toe tectum (MBT) of the higher visual center of fowl to
photostlnulation 1n ci;1clt embryos.
, study Wa6 maue on 95 ch1ck eQbryos of the russian white variety from
IG to :1 davrs or incubation. For ot the b10potcntio.ls frOltl tr.e
surface we the proceUures. We dissected a sJIlAll sectior of
the abo're the head of the embryo, cleared the subdural membrane with m1n
ertU. oil, and then cu.t the eI:1bryonic Il'embranes. We extr&e't.ed tbe head or
the em'tryo frOM. tne raised it above the level at the shell and fixed
it in this position with a immobilized the embryo vith an
injection of D1plac1n solutiDn (O.3-0.
L
We ex
t:.e dorsal. surface ot the ontic lobes of the midbrain UDder a binocu
1u tr.iCT05COPC. ve relllOved the <iura t18.ter and arachnoid. We J'llaced the
ez:tbrJo irl a chamber wUh constant telll:oerature (38C) anei hUll1 d.1 ty. For re
cording lhe electrical activity ve used electrodes or thin silver vire
vitoh a s!lhere at one end (sphere cllameter 0.3 1lIll.). The electrode. vere at
tached to zprinr..a vhich usured contact vi tb the brain. \Ie attached an In-
di rrererlt. needle electrode to the rear muac:les. neeordinp; of the electri cal
Qct1vity VQS r..ade on a 1nk-writing electroencephalograph of the
type vi th 8yqJet.r1eal input. (Frequ.ency tranall11ss ion band t'rorn O. J
to 100 t:z. aaxUl\D senllt1vity or channels: 2 microvolts 1 llII1 of re
cording).
rhythmIc and palreJ flashes of from a l'hotosti
mulator in the frequency raDRe tram 0.1 to 30 H2. Flnsh time wu 30 micro
seconds. In stimulation, the lenr,th of series of
vas lC-l; Tile interval 1etveen tvo wac one minute.
-2
lap of the paaowat1mulator vu CIlI IlVIlY th<: r.".!<:. We the
experilllent roO earlier than ?O minuten nrter e'J"t.ir.l.;uishlnr Ill] H/:ht In the'
chnmbcr. Art (EJCO) vas ur.eC: 1.0 "'0111 tm' nt'lnTl:'.l vi \.(.1 'l.ct.l
vlty of the e-.bryo.
P.I:rmLTS OF TIlE 'rr.3TS
StiaulaUon ot by a Bingle did not. f'voke any
ehanp. in the bioelectric activit)" of the MBT or the chick prior to
the 16th dq or embl")"onic Ure. Froa thia time on, and before hatchine, ve
recorded the ncked potenUala (!:p) at vadows pointli of the 40raal surface
or tne c:ontralateral IlBT in reaponee to a I'I1ngle ,motoltiTrlU1us. 7hey vere
characterized by a specific 8IIIOuot or amplitude, .hape, oolarlt)" of cOl!lpO
nent., et.e.
The eYOked potential in l8-day old chick embryos conslst.ed of three
componenta: a surtaee-poaitive oscillation ot quite
811p11tUde, Il nesathe cOIIlpOI1eDt follovlng 1t and a slow posHhe vave
UTe 1, Di. In a number or teat., dependlnc on the presence of early or latc
poaltivity, it Val to record m'no- or double phaRe Ft alonr,
the triple-phase::P. Th@ negaUve cClllronent vas more constant and sharyly
exnreeled. In lIloJst cases the NIlpl1tudl" of conpor.ent
the amplitude ot the positive vave follovinr. it by 2-3 In this case
the duration or the negatlYe component was several shorter thftn du
ration or the vaYe.
One of the chara.cterhUc features or the ::;P 1n embryos is its
great variation of EP amplitude and duration of which occurred
in the course ot the teat even in the ssoc embryo, not to speak of the grcnt
Indi vidual varl abi 11ty ot these lUllOWltS. In most of the thO! tude
of the negative component tluctaated tron 20 to 20 while its du
ration vas trOln 50 t.o 70 microlleconuti. Another chnracteristic
or chick etr.bryo r;p 1n the given perioJ of deve10mcmt vns t:le u:lstel\uineos
ut their app8araaee. 01't.en at the atart of the tests llnht flashes \lid
not evoke &nT changes in the electrical actiVity of the !!DT. Then, (LS the
stiMulation vas continued, EP's or smAll amplitude to be reeorded (on
the order ot 10 microvolt. (Figure 1, A-D)). rrndually their in
creoaed and, reaching a certain specific value, at this ror
minutes or tens or alnutel on the teat cond1 tic::.a.
In ohlttlng to the
response to each flash vas recorded to limit 1. C_
9
).
For 18-d"Y old embT)'OlJ this limit vas /I, frequency or 5 t'lBShes TIer second
1, F). In this way the nature of the PB? reaction depended on the
t'requeney ot the Btlnulation used. With cCllllParaUvely rare st.iJllUlation
reaction eonaieted ot typical evoked resronaea vith sharply expressed
nents. Wi th an increase in the frequency or the 11ght. slow
vaye. bet;an to prevail. Wi tn a rurther incr@..e or the rapid
-3
d

A ' t"V
, 11-" sec
B I

, $
I

On no. III 0' ,_, 11 100 to., 1 f I
0'''''' ,
pp " 0
, ""',.' 4ft .." 1'$ ,tI_dt' .....
,"
au to b-- .. .t.,tIlt
1. P.loelectric reac
tions of the of 1
old chick eMbr,yo to photo
st1Jllulation
of stimulr.tion
(numher of fllUlh('1I
second): A and L -' 0.2
indirect continuation of A);
C - 1; - 2; - 3; F -
r, - 10; l: - !laind fleshes
with interval of 150, 300
and 100 microseconds. 1
of electrical nc
tivity ot' 1 -
[The upward movement or the
pen to
vity ot actlTe
responses began to appear again, 1!ULinly of nep:ative eomr-()nents
of EP (Figure 1. A-F).
In res!'onse to tbe use or 8. fliekef'lng 111")lt vi th a frequency or In
flaz:.es a second or more. a certain exaltat10n or compression of bacltr;roun,l
activi ty vas observed which vas not aeeom!,af.ied by 13orh:rthM.1c res!,onsec.
Orten on-type and. less rarely. orr-type relponses arORe in resnonse to nwitch
Ina on the st1aula't1oa.
In old elllbr;yoa the LP were by a 8Illplitu(]e
and a remanent mani fes t ation Indi vi dual va.riability. althoup,h observed. vaa
not as 3ignU'icant &8 1n lE-cia)- olto cIIlbry08. The ampli tude of the
negative vas 50-100 aierovolts (up 20n in isolated
duration - about 50 Ill1croseconds. The shape of the Er and the rolarl ty
of its cex:lponenta did not differ trom thOle in the If.-/le.y old
2. A).
,
.at -+ +
4 4
A
,1.:, He
.. ,
.1
.. ..
"
I
_.
s , 1 ,
II
S 00
v

,
III
2. Dioelectrlc reac
tion of ttBT of 19-da.y old
chick mcryo to !>hotostilL
ulation.
Frequency of stimulation
(number of nashes ner
second): A 0.2i P. - 1.
C - 2. 0 - 3; E 5. r
10. n - paired flashes vith
intervals of 200 and 100
microseconds. Other
boll sane as 1n 1.
,,"w'.bI.t 1'1 ..... 14

.. ----'l
o ,

.:.....-----+------.....-
Continuous stimulation ot the eye by light vith a frequeney
or 0.1 per second. continued 20-30 Ildnutel. did not evoke a noticeable re
4uctlon of EP eapl1tude. Hovever. EP 8IIlP11tude did not renain strictly
c:onltant duriD!J the entire period ot stiJllU1ation: it increased and decreased
periodic:al1. altbough to a very inaip;n1ticant <leeree (by no !!lOre thl\
or the in!t1al uount). f:tll11U1at1on or the eye vi th a f'rectuenc!y or ().) '!"CT
second tor 10 lIlinutes also evoked no reduction of re9T'lOnllC ampl1 turle. !:ov
eler. at this frequency or nlckerinr; of fallin:,: out of crrtnin
EP rran the series ot isorhyth:nlc reS!'ODses vas observer!.
The _turf" of re9ponses of 19-day em1;.ryos to rh:,-thlr i c l1i';ht Itiaul.1
vU.h & trequeaq ot 1-5 per .eooD4 did not Utter troa t.hat ot the 18-d.q
embryos. Tole only distinguishing feature vas Do. shuper reproc1uction of rhy-
thin at a stimulation of 5 flashes per sp.conu 2. A
t'rcquency of 10 !>Cr second vas not reproduced 2. r-). In
certain cues. the rhenomenon of rh:Ithm t.ransformat-ion oec:urred at S tlJll
wation frequency. A rhythm or 5 nashes seeond vas \Wed :s nstead of 10.
-5
C ' __ _
A 't I
I t.::., Ite
B
.. , .... ..... ...... l ..... 1 ::-i"',;:'(' '.. . .:Ol.lcC'tri:: '{',.
act,ion of ?o-d,v 01
r
1 ('\,1
etir,!":rc to nhot03t1Jnulnt1vn.
t l
or
c , I-+'-+I-+ l _
or
A - 0.2; - 1;
C - 1; D - ?; r. _ v
_ f ---IJ,JJ,J;J),J,J,lJ,J,I,J).ItJ)J,J,J,J,IJ)J,J)AJ,
lr.. - Other
I)
sane in 1.
, _---lllo6l.&.I... ... ....... +,,11104'''''.. ..... ---:- I ... l+I+' .... I... 1+
,
.- ' __"'''IllI' _
, lllll'
t ""',J".I. ...
W
'l_.......'....
p
, __-4 _. ' '''._''-

G 1 __-:-- -..----
Figure 3 shows iili'r' respon to stimuli or var,,-ing treQuenc:. lr,
ar. embryo ..tter 20 ot incubation. A characterist.ic of F.P eTOke4 b7 a
single Tlhoto.t1mulus IS .. re.1d\&&l cha.rge, vbieh consists of a R1"OUl'l or Slow
vaves that tollow EP (ngu.re 3. A). In IlIOBt cues t.he az:splitude ot the
negative EP cCDJ'lOnent exceeded 100 microvolts. In stimulation or the eye by
a tlickering 118ht vtth a frequency or 3 flashes per second the :mT reaction
uau&ll:r eoaa1sted of tY1'lIc:&l PI. at each fluh. in contrast to the '.tnT re
action or 10- and 19-day emb17oB, which vu exnressed prll'l.!U"lly by slov
(Figure 3. D). In response to photostimulatlon vi tll a freet-eney of 10 flas:J
.s per lecond a rbTthla or 10 per aecond vas recorded in the :.mT. but i:: the
maJorl t;y ot instanc the index or ....iJ!li1ation vas no than O. St1z:1
ulaUon at a frequency ot 15 per second evoked the on-reaction phenanenon
(Ie.. otten. the off-reaction) and did not lead to any ch&nfl':e of back4round
(figure 3, G).
-6
In etlaulatlOil by pa.1re4 l1ght fluhes. aeJ)a1"&ted troIIl one another
'by intenals ot 3 ueollc!s to 50 aicl'Oleeonds. the relponse to the aeeoad
fluh eU.appeared at a certala crltieal interval between st.imuli.
cbanged vithin ot 60 to 200 aicroseconds on a number
or ultterent eoacl1t101l8 1. Hi 2. 0). Anal7sh of the data
ab0ve4 that. lD41'f'1clual Yariat10u of this ftlue are IIIOre a1gn1t1eant in coa
partlon to IP t1uetuatiou ot tbe retractive !'eriod.
DISCUSSION
StudT or the bioelectric reactionl of 41fterent aeetlonl ot the
Yiaual to 11lbt stbluli 1n chick BbryM vu conducted b7 Peters
et ale (11). Howftr. In this york he U8ed an 1aproper lead (ld4bra1n - cere
bellum) the e.bryoa " ... DOt l.-obllized. the electrodes ve1"e lneened to
a 4epth ot 3 _. which cornsponded to different MBT layera in embryo. at
YV!oua per10da ot 4eYeJ.oJ-At.. Theretore." attempted lID InveaUatlon
vb1eh vould Dot. 1llclu4e th..e M'thodologleal lrwlequaeiea. In calP&rtng the
data traa our experiment. vith those of Peten 1t turned out that. along
with certalD cUfferenee there V&l a c:orrelJlODdence ot the buic results.
POl' lnatuee. deteJ"ll1na'-1011 or the t1JDe of deftlopeent or the _bryo. when
MB'1' relpanae. to lipt .t1auli appeared (18 dqa ot 1neubation). TarlabUi ty
of VP llADit.. preT&leDCe ot tbe coeponent and eatablish
!lent ot u upper l1a1t for repl"Odueed frequenciea of light flickering (10/
.econd) coatlna Petent flncUllS.
Ulltortunate17. tor Peters' work. we did not find any Itudies
ot the proee.. or the tuDetlOD&1. maturation ot the visual analyzer in towl.
Hovever. a certaln congruence 1n the atructure ot fowl and the neocortex or
'ermlls (12). tne pre.ence of aix sharply pronounced 1.,erl. gives UI
a buis tor dravlna certain puoallela to reault. of ont.ogenetic It.udies of
the rtaul area ot the cortex In MIII'.al.s. ot the literature shows
that in ua1llaU 'bom blin4. (rabbit. cat) the Ell in the vhual zone of the
cortex appears duriDS the tirst 4Al.Ys ot postnatal 11te. Thus. Harty [5] 1e
acr1bea th. appearance or the EP In relponse to a1l1Rle nuhea on the
2-5th da.Y ot poetDatal deftlOlll8Dt in kittens uad at the end of the tint
ft_ 1n rabbit.. ID the work of faneI' [8] the EP ill reaponae to 11Sht tluh
ea v.. reeol'4ed In kittena and rabbita CD the 3-5th" _ arter birtb. Ell1D1
SOD and obaenell EP appearance 1n relpoue to single pbotoatlmuli In
ki ttena trca the eighth dIlf ot po8tnatal lite. In nevbol"ll children the r;p
reaponae to light flube. la recorded during the f.irst houri or Ure. In
a4ditiOil. EP can be recorded nen In J'l't!IIl&ture children 1300 gr8r.lB
on the second _ arter birth [10]. Thi. tact c:oatinlUl the hi,,h of
tuDctional aatW'lty of the rt.ual analyzer 1n newborn children in eClllJ)&l'l
.on with &Dial. bam blind.
The frequency r-ange vlthln vhich i80rbYthmic or the visual
centers are recorded is .000000t videI' In chick .wryOI than in the represen
t.atift an1a&la bieh were .tucUed. The capaclt7 to reT'roduce a rhythm of
-1
light tlickeJ"1n8 of per second appeus 1n cata and rabbits onlr on the
22-30th of postnatal deftlow-ent [13], while in newborn children the
upper limit 18 a tnqUIDCJ of 3-6 per second [9] or 3-5 per second [10].
Our data confirm the ot the visual in
chick embl708, ud are 1n direct correlation vith llI01"!lhologie&l .. will
have sbovD a quite high degree ot ..turity ot the studied sections of the
visual 8Ilalyzer in chick eaabry08 in the tinal d.,ys of prenatal development
[llt-18] Blocb-.1cal studies by Linc1eman [19] shov that 'by the 19tb of
incubaUDD the acetylcholine COI1te11t .. vell .. the cholinesterase and al
kali.. " ..... the retlna Incre..e sburly. Thus, 1n chick
_bl'7Os, b7 the IS-19th of incubation, a definite critical period in de
begina, a period by abrupt biocheJ:11cal, structural .
uel tlmctloaal cbarges, vh1ch sbow an 1ncreulag degree ot _tunty of the
nsual analyzer.
Certain researchen [5, 6-10] noticed a rapid adyance or fatigue of
spee1!1c reaponses of the "f1sual and auditory are.. of the cortex during a
cClBpvatively Slow rbtbm ot stiaulatlon (0.3-1 p.r second) 1n newborn chil
dren u well as in rabbita and eats in the tint dqa ot postnatal lire.
The results of our tests are contradictory vi th respect to the ci ted
data. The Yisual centers in the chick eI!lbryo are capable or responding to
each light fluh at a st1aulaUon frequency or 0.1 to 1 per second with
continuous .t1lllulation in the cour ot several lo-a1nute J'l4!rioda without
significant reduction of EP .-p11tude.
The considerable individual Yariation in the size and shape ot the
EF, noticed b7 u.s 1n tbe lut dIt. of embl')"Onic deTelopment, is in a,o;reenent
vi th tbe data obtained by Farber [9], Hrbek [10) ud others. However, on
the backp'ound or tbi. Yarlabll1t7 one can detect a tendency ot EP 1IIIIl'11
t.ude to increase in the cOUl'le or dnelopDeDt. Further, in 'j)asl1ng troe the
18th to 19th dtti1 at de-felOJMnt there is alia an expansion or the rang\! or
paired frequencies. 'rhea. dat.a al.o that b7 the 19th ot em
bl'Jocic: the Yin&! anal7ser at the chick "ryo 11 well-developed
tuDet10na1l.
We could find only oae work (Farber [8]) vbich treated the cycle or
scitabll1ty or response. of the ri.ual area ot the cortex 1n eat. and rab
bits. In 7-U-_ old ulJ11&18 the absolute retract-lYe vu 120-200
micro.econds. In our tests the value or the absolute refractive reriod fluc
tuated trca 60 to 200 IIl1crosecoada, depending on various caDdi tiona. These
are entirely c<apuable resulta.
In sp1te ot the difterenee. in the organ1zation of nerve centers
which lIl8ke up the visual analyzer of mam- and towl, there are a number of
'
ties 11. the proee.s at the twletional developll8nt ot the giyen struc
tures which are ec:.DOn to representaUves or the.e tvo cluses.
C<liCLUSIOUS
1. Potentials evoked at.1nu.le.tion of the by single light
flub Call 'be recorded h'ca the dora&l surrac:e ot the I11dbra1n tectum (MBT)
of tbe chick -.bryo. besinn1ng on the 18th of embryonic
2. Thne potential. have a three-phe.ae shape (positive-negative
poa1ttYe) vith preYalence ot the Degat1ve coaponent. In a number or tests
the initial or later polithe ccaponents may be absent.
3. Aga.1nat a background of a signltlc:ant variabll tty at Ell e.r.rr1i
tUde there h a tendenq to increue the response uapli tude in the course
ot cle-.elopaent.
4. In stimulation or the e,yc by flashes with fre
fluency or 0.1 to 30 per second. i.orhythmic: re8J)Onaea up to a frequency of
5 flub.. per second 1n le.. and 19-11ay old embryos can be recorded and u-p to
10 nub per second ia 2O-dq old embryos.
5. The absolute retractive period ot response during stimulation
or the by paired light nashes varied t'r<lIIl 60 to 200 microseconds 1n
_bryo. aner 18-20 de3s at IncubaUon.
(;. Theae data abow tbe quite high degree of functional develoJlr'lent
Of the visual analyzer in chick eDbryoa durine the tinal ot Jlrenate.l

m.FEREHCES
It! ... E. H & a. S. G 0 14 r 1111. EEG.. CUll. Neuophpiol.. J. 4eS
('.'). - 1211.-T. , Acta "piol. IIA. 2t. 5t - 131 R. J. E Ill.,.
... alC. ao CUa. . tI. 863 (1110). - (4) G. H. G I .. u
.... II. I" M.... 1.--. c..r.. Ser. 16 17. 30 (tMn. - 15) R. )( r' J .
...... " ....... lla ..1" .. lie l'Um.",,, Parb (1182). - II) B. n. E
........ VI ..,... _1*7 ......., -.
-. ADB PCeCP. )(. _ (till). - 17 IT. r. y,. a I r. H. 1J
a .. ...... Kiln ....,......... ..,... AIOI
CDCPt. e,.,... 71 (t..,. - ,I) ,A. A. Ch php. B
. ......... &10...... Jba, ADK PC:P, _(I .
I'.-AI '- eu.oa.. CCCP.IO7 (I.n. - ItOI A.. K r i. I
a. CIa........ .. I'. 575 (111'). - Itt I J. J. It. \. "I A.. 1\. V
I. T. If. e". r I, J. Ell"'. Zool_. 4St (1111). - 12) G. C. H. r
a. Po. C r 01 '" J. CoaMr. JIf..roI _. I (t..,. -I UI R. J. E IUa e'
.. a. c. Wile 0 H. J. )f".NIlIapfoL. D. 3IS (tl8O). - It41 S. a. c. j a1.
I. lat... At. " l'taYlt.... a. 3!7 (ta"). - lUI J. r. T." Tnv. !alt .....
... bioi. U.lvWHrii. II. 1 (1923). - (III A. w. W " ..... W. S. B. r ..
A... Nataral"'. 41. lit (tlOl) .- 1t7J A. ye. G. ta ell' D, La Cell It I.
C
t
ll2). - Ull D. R a I c II I. H. Wa 1_ c Il. I. tBlodMa1a&r7 of
Nen_ alMe It_>. - (I') U....... ..,.p. DO: A. R...Dan.n. A....
.....,.. II.. YOlk ct.
DOC!'J8LM n VIIl tlK
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