M.N. Myrzakhanova
Kokshetau State University
named after Sh. Ualikhanov
Spontaneous and evoked
contractive activity of lambs’ isolated lymph
nodes in ontogenesis
It is a well-known fact
that in the course of the animal world’s evolution humoral regulation of the organism
functions turned out to be much older than nervous regulation. While
investigating the lymph nodes of bull’s mesentery it was found out that all the
tested concentrations of antibiotics in different ways inhibit the contractive
activity of lymphatic vessels [1]. Serotonin is the morphogenetic factor
involved in the regulation of brain development, cardiovascular system and
other target organs. The morphogenetic influence of serotonin makes itself felt
at definite for each target critical periods of altricial rats’ ontogenesis in
pre- and early postnatal periods [2].
According to the
results of the researches, conducted on the bulbospinal specimens of the brain of
18-20 day-old fetuses and newborn rats (0 – 3 days), in the conditions ‘in vitro’,
the development of the central mechanisms of the respiratory rhythmogenesis is
accompanied by decreasing variability of respiratory rhythm, increasing duration of respiratory discharge
and redistribution of power capacity in the spectrum of the electric activity.
Adrenoceptive neuronic structures of the pons have a tonic inhibitory influence
on the respiratory rhythmogenesis of the fetuses and newborn rats which is
modulated by the chemosensing mechanism [3].
In the organs of newborn lambs it is found a
high content of adrenalin – 5,92-136,81, noradrenalin – 37, 41 – 2018,36 (ng
per 1 g of tela) and a less quantity of catechol amines. Consequently, we may
say that newborn lambs have relatively perfect organization and activity of the
thyroid gland, atrabiliary capsules and pancreas. Such distribution of hormones
by their content corresponds well
enough to their physiological role and the degree of sophistication of the structurally-physiological organization of
the organs[4].
The reaction of the
contraction force of rats’ myocardium was observed at the age of 3, 7, 14, 21,
100 days old in response to serotonin under adrenergic block and in the
conditions of sympathectomy. It is
defined, that the block of adrenergic receptors modulates inotropic effects of
serotonin: it amplifies it at the age of 3, 7, 14 days, leaves it unchanged at
the age of 21 days and reduces when the rat is adult. Serotonergic system at
the early stages of ontogenesis plays the leading role in sustaining the
inotropic function of the heart if compared to the adrenergic one [5]. In
innervation of the lymphatic system a great role is played by the sympathetic
nervous system; and there exist a number of works dedicated to the influence of
adrenalin on the lymphatic vessels and nodes [6].
At the present day
spontaneous contractive activity of the lymphatic vessels is found almost in
every animal [7]. The question as to whether lymphatic nodes are involved in
the lymph circulation is today of a particular interest. This is explained by
the fact that lymph nodes develop in the organisms of those vertebrate animals
who have lost lymph hearts [8]. The task of our research was to study the
formation and development of the contractive activity of rats’ visceral and
somatic lymph nodes on the 1, 5, 10, 15, 20 and 30 days after their birth.
Research methodology
To carry out the
experiment there were taken as many as 60 laboratory rats and 40 ground
squirrels. There were taken from the rats 17 isolated specimens of
submandibular, axillary, inguinal, cardiac, intestinal, mesenterial, nephritic
and hepatic lymph nodes. From the ground squirrels there were taken 7 isolated
specimens of submandibular, axillary, inguinal, cardiac, intestinal,
mesenterial, nephritic and hepatic lymph nodes. Physical inactivity of the
animals was secured by means of ether-chloroform anaesthesia.
In sum, there were carried out 820
experiments with the isolated vascular specimens of the nodes. Spontaneous
rhythmic contractions of the lymph nodes were made on the 1, 10 and 30 day
after the birth of rat litter and on the 1, 15, 30, 90 day after the birth of
ground squirrels’ litter. In the course of the experiment with the rats’ and
ground squirrels’ isolated lymph nodes their contractive activity was explored;
prosected nodes were placed into thermostatically controlled chamber with the
flowing solution of Krebs; tracing of the contractive activity was done with
the help of the mechanotron 6Mx1C, in accordance with the established
procedure, graphical recording being done on recording meter H-327-5 (Orlov and
others, 1975; Luchinin, 1979; Myrzakhanov, 1987), on the unit modified by
Hanturin in 1996. There were used longitudinal specimens of the lymph nodes,
from 5 to 10-12 mm in length. One end of the longitudinal specimens of the
nodes was fixed to the bottom of the chamber of the vertical type or to the
sidewall of the horizontal type, the other end was attached to the force
sensing device (mechanotron for the sensitive type 6Mx1B). For the isolated
lymph nodes of the lambs’ there was used the solution of Krebs of the following composition: NaCl –
124,0; NaH2PO4 – 1,2; RCl – 5,9;
CaCl2-2,5; MgCl2 – 1,2; NaHCO3 – 15,5; C6 H12 O6 – 11,5 mol/litre of
the distilled water. In the course of work there were used solutions with pH
7,2-7,3, at the temperature of 37° C. Nutrient
solutions were oxygenated with the gas
mixture: 95% O2 and 5% CO2. The following
physiologically active substances were employed as irritators for the nodes:
adrenalin hydrochloride, acetyl chlorine chloride, noradrenalin.
Contractive
activity of the isolated lymph nodes was registered on the graph paper of the
potentiometer KSP-4 or milliampervoltmeter of the tracer H327-5. During every
experiment there were provided conditions for life sustaining of the isolated
nodes during each experiment there was followed a certain order of making
intervals of agents injecting and the order of the substances washing off.
Results of the research.
Adrenalin in
submandibular, axillary, inguinal, mesenterial, intestinal, hepatic, cardiac
and nephritic lymph nodes of the
postnatal development of lambs makes on the 1st day from 1,37±0,0152 ml/min to
1,99±0,0210 ml/min; on
the 30th day up to 3,5±0,0405 ml/min; on
the 90th day 3,4±0,0381 ml/min and
on the 180th day up to 1,83±0,0203 ml/min if compared
to the initial level. The minimal effective dose was on the 30th day in the
submandibular nodes – 39%, axillary nodes – 30%, and was 34% on the 90th
day; cardiac – 45%, intestinal, mesenterial – 35,7%, nephritic – 36,4%, hepatic
– 27,2% and on the 90th day was 30%, the dose in the inguinal nodes
– 28% and on the 30th day – 30,6%. The maximal dose was on the 1st,
90th and 180th days in submandibular – 68%, 40,3% and
74,8%; axillary – 65% and on the 180th day - 46%, cardiac – 93 %, 74,8%
and 90%, intestinal, mesenterial – 72%, 40% and 55%, nephritic – 88%, 45,5% and
63,5%, hepatic – 96% and on the 180th day – 40% and inguinal – 61 %
and on the 180th day was 40%. (see picture1)
Picture 1. Contractive activity
of lambs’ isolated lymph nodes when
reacting to adrenalin
Contractive
reactions of the lambs’ lymph nodes to acetylcholine made on the 1st
day from 1,36±0,0151 ml/min
(submandibular, cardiac, nephritic, hepatic), 1,40 ±0,0156 (axillary, intestinal,
mesenterial, inguinal), on the 30th day from 2,26±0,0251 ml/min to
2,95±0,0327
(submandibular, hepatic), 3,9±0,0433 (axillary,
intestinal, nephritic), on the 90th day from 3,56±0,0396 ml/min to
2,95±0,0327
(submandibular, axillary, intestinal, nephritic hepatic, inguinal), 3,5±0,0406 (axillary)
and on the 180th day from 2,96±0,0329 ml/min to 2,95±0,0327
(submandibular, nephritic), 1,96±0,0218 (axillary,
hepatic, inguinal) compared to the initial level. The minimal effective dose
was characteristic on the 90th-180th days for the submandibular
lymph nodes – 29%, 34%, on the 30th and 90th days for the
axillary nodes – 27%, 30%, on the 30th day for cardiac – 30%, on the
30th and 90th for intestinal – 29%, 33%, on the 30th
and 90th for mesenterial – 30%, 35,8%, on the 30th, 90th
and 180th for nephritic – 27,7%, 30%, 36,4%, on the 30th
and 90th for hepatic – 35%, 27,8% and on the 30th and 90th
for inguinal – 32,3%, 33,2%. The maximal dose was on the 1st and 30th
days in submandibular nodes and on the 1st and 180th days
in axillary, intestinal, mesenterial, nephritic, hepatic and inguinal; for the
cardiac nodes of the lambs of the postnatal development it was on the 1st,
90th and 180th days.
Picture 2. Contractive
activity of lambs’ isolated lymph nodes when reacting to acetylcholine
Histamine influences
the contractive activity of the isolated specimens of the lymph nodes.
Specimens of all the enumerated lambs’ nodes developed dose-dependant tonic
contractions responding to histamine. Histamine in lambs’ submandibular nodes
made on the 1st day from 0,93±0,0103 ml/min (inguinal) and
on the 30th day from 2,30±0,0255 ml/min, on
the 90th day from 3,38±0,0375 to 3,62±0,4025 ml/min, on
the 180th from 1,73±0,0202 to 3,26±0,0362 ml/min if compared
to the initial level. The minimal effective dose was characteristic on the 30th
and 90th days for axillary nodes (25,6%, 30,1%), cardiac (30%,
28,7%), intestinal (32,2%), mesenterial (19%, 2,8%), nephritic (28%, 30,8%),
hepatic (32,3%, 35,6%), inguinal (18,2%, 20% on the 180th day); the maximal
dose was characteristic on the 1st day for all the nodes and on the
180th day for intestinal,
mesenterial, axillary (43,5%), nephritic, hepatic and cardiac (50,7%) lymph
nodes of the postnatal development (see picture 3)
Picture 3. Contractive activity of
lambs’ isolated lymph nodes when reacting to histamine
Serotonin in
submandibular nodes makes on the 1st day from 0,07±0,0008 to 1,27±0,0141; on the 30th
day from 1,32±0,0146 to 4,21±0,0468; on the 90th
day from 2,20±0,0244 to 4,23±0,047 and on the
180th day from 1,33 ±0,0147 to 2,60±0,0289. The minimal
effective dose was characteristic on the 30th, 90th and
180th days for submandibular (26,5%, 11,6%, 13,4%), cardiac (3,44%,
2,75%, 4,34%), intestinal (23,5%, 26,6%, 38%), mesenterial (16,5%, 23,2%,
30,5%), nephritic (12,7%, 13,3%, 25,5%), hepatic (12%, 15,4%), inguinal (24,7%
on the 30th, 90th days) lymph nodes. The maximal dose was
characteristic on the 1st day for all nodes and on the 180th
for axillary nodes (91,3%, 41%) and inguinal nodes of postnatal development
(81,8%, 50,2%) (picture 4).
Picture 4. Contractive activity
of lambs’ isolated lymph nodes when
reacting to serotonin
Picture 5. Alterations in the
contractive reactions of lambs’ isolated lymph nodes in response to adrenalin.
In picture 5 there
are given alterations in the contractive activity of the isolated specimens of
lambs’ lymph nodes when reacting to adrenalin. It is seen in the picture that
against the block of b - adrenergic
receptors the quantity of contractions of hepatic, cardiac, nephritic nodes increased
on the 1st day reacting to adrenalin. The level of the contractive
increasing is the biggest on the 90th day for cardiac nodes and on
the 180th for the cardiac and submandibular nodes. The level of contractive
increasing is the lowest on the 30th day for the hepatic and
inguinal nodes.
Another set of
experiments was dedicated to the study of acetylcholine influence on the
contractive activity of the isolated specimens of lambs’ lymph nodes. In
picture 6 one can see graphic presentation of the contractive activity alterations
of the isolated lymph nodes reacting to acetylcholine
Picture 6. Alterations in the contractive reactions of lambs’ isolated
lymph nodes in response to acetylcholine.
As it is seen from
the picture all lymph nodes of newborn lambs, except cardiac ones, have the
same frequency of contractions. On the 30th and 90th days
one may observe passive character of nodes, which is caused by the influence of
the extralymphatic factors. After 180 days one may notice a considerable
increase of inguinal, axillary, cardiac and hepatic nodes and decrease of
submandibular and nephritic nodes which is typical for adult lambs. However,
the range of alterations of nodes was insignificant in comparison with the
adult animals.
Picture 7. Alterations in the
contractive reactions of lambs’ isolated lymph nodes in response to histamine
In picture 7 there
are shown isolated specimens of lambs’ lymph nodes reacting to histamine by
dose-dependant tonic contractions which are realized through H1-
histaminoreceptors. The degree of receptors’ affinity to histamine is the
biggest in the lymph nodes of the 1st day of life, then, in
decreasing order follow nodes on the 30th and 90th days,
and on the 180th day frequency of the node contractions increases.
Spontaneous tonic
contractions are typical for newborn lambs, since the age of 30 days there
begin to appear different rhythmic contractions. At first there appear single
waves (rudiments of phase contractions).
Picture 8. Alterations in the contractive reactions of lambs’ isolated lymph nodes in
response to serotonin.
Isolated specimens
of 1-3 month-old lambs showed the existence of phase contraction periods of
various duration. These contractions differ from the picture of adult lambs’
contractive activity in a high frequency and low amplitude. The research showed
that phase rhythmic contractions of the lambs’ lymph nodes (picture 8) develop
on the 1st, 30th and 90th days of the
postnatal ontogenesis.
Discussion.
The given results show
postnatal development of the receptor area of vascular wall of lymph nodes,
gradual increasing of its susceptibility to vasoactive substances on the 1st
and 180th days. Besides, we discovered on the 30th and 90th
days decreasing of the contractive responses of the nodes when reacting to
adrenalin, acetylcholine, histamine and serotonin in the course of lambs’
development.
The dynamics of the
formation of the contractive activity and quantity of the contractive responses
of the growing animals’ lymph nodes proceeds in a parallel way with the
development of smooth muscle elements in its wall. As showed N.A.
Akhmetbayeva’s and her co-authors’ researches (1980), L.E. Bulekbayeva’s research work
and N.A. Akhmetbayeva’s work
(1982), the formation and aging of the smooth muscle elements and other
structural components of the thoracic duct of dog cubs is completed by the 3rd
month of their life. These age-dependant morphological alterations correlate
with the formation of the contractive activity and evoked responses of the
thoracic duct of dog cubs.
It is shown, that
the reactions of the lymph nodes by the periods of the postnatal development
outrun tonus changes of the growing animals. The same lateness of lymph node
development is also found in the prenatal period of mammals’ development.
Thus, the received
experimental data show that the formation of the contractive activity of lymph
circulation mechanisms occurs both on the level of the regional adaptation of
lymph vessels, and on the level of selective adequate changes of the lymph node
activity in the period of the postnatal development.
Literature:
1. М.В. Андреевская.
Влияние антибиотиков на сократительную активность гладкомышечных клеток
лимфатических сосудов. Т.1. С.140. I – съезд физиологов СНГ.
Т.1. С.140. Сочи, Дагомыс, 2005.
2. Д.И. Насырова, А.Я.
Сапронова, А.В. Балбашев, А.К. Нанаев, М.В. Угрюмов. Развитие центральной и
периферической серотонин-продуцирующих систем у крыс в онтогенезе. I –
съезд физиологов СНГ. Т.1. С. 166. Сочи, Дагомыс, 2005.
3. И.В. Мирошниченко, Е.А.
Зинченко, А.Р. Ахметзянова. Развитие механизма дыхательного ритмогенеза в
перинатальном периоде онтогенеза. I – съезд физиологов СНГ.
Т.1. С. 167. Сочи, Дагомыс, 2005.
4. В.И. Максимов.
Становление в онтогенезе эндокринных функций. I – съезд физиологов СНГ.
Т.2. С. 128. Сочи, Дагомыс, 2005.
5. А.Ф. Ахметзянова, В.Ф.
Ахметзянова, М.И. Гарифулина, В.В. Кириллова, Р.Р. Нигматуллина. Взаимодействие
адренергической и серотонинергической систем в регуляции инотропной функции
сердца в постнатальном онтогенезе крыс. I – съезд физиологов СНГ.
Т.2. С. 129. Сочи, Дагомыс, 2005.
6. Рейдлер Р.М.
Распределение катехоламинов в тканях кролика в онтогенезе. –Журн. эвол. биохим.
и физиол., 1968, №4, с.55-57
7.
Булекбаева
Л.Э. Сравнительная физиология
лимфатической системы. Изд-во
«Наука» КазССР. Алматы,1985
8. Мырзаханов Н.М. //Сб. междунар.
конф. «Проблемы лимфологии». –Новосибирск, 1987. –С.78.
9.
Орлов
Р.С., Борисова Р.П., Мандрыко Е.С. Сократительная и электрическая активность
гладких мышц магистральных лимфатических сосудов. – Физиол. журн. СССР, 1975,
т.61, №7, с.1045-1049
10. Лучинин Ю.С. К анализу
спонтанной активности лимфатических сосудов собак. – тр. ин-та физиол. АН
Каз.ССР, 1979, т.24, с. 38-49
11. Мырзаханов Н.М. Роль
лимфатических узлов и сосудов в
продвижении лимфы. –Вестн.НАН РК. – 1994(б). - №3. –С.70-76.
12. Хантурин М.Р. Эволюция
транспортной функции лимфатической
системы: Автореф., дисс. док. биол. наук. -Алматы,1996. –С.32
13. Ахметбаева Н.А., Рейдлер
Р.М. Адренергическая иннервация грудного лимфатического протока у собак в
постнатальном онтогенезе. - Журн. эвол. биохим. и физиол., 1980, №2, с.207-208.
14. Булекбаева Л.Э.,
Ахметбаева Н.А. Морфо-физиологическая структура грудного лимфатического протока
у растущих животных. – В кн.: Проблемы функциональной лимфологии: тез. докл.
Всесоюзной конф. Новосибирск, 1982, с.30-31.