03kulijm
<r\í
IL   O <^3
a) HoubovcJ
ti) Zariavcí
Pseudocél
Trávicí trakt
^^-g?
c i H is-:ice
Coeíenteron
Srdce                             .'crzári
\      Ostíe               céva

Střevo -errosé
c} Hmyz
Ostie             Sfdce            -išncce
\      _7
^l^S?
Tepna
e) Korýšř
Dýchací
Tepna     Srdce            kapilární 3íf
Hemocél
ľ) Měkkýši
Laterálfií          Dorzální       Cělamová
srdce                céva               dutina
Kapilární sít"
Komora
Dorzální        Kapilární
"Célom
g) Knoužkovci
Lymfatická   předsíň   Lymfatický   Žíla spojka                             kanálek
h) Savci
Otevřené a uzavřené sst.
1) Rozdíl mezi systolickým a diastolickým tlakem je malý. 2) Periferní odpor je malý, s čímž souvisí i malá intenzita srdeční činnosti (výkon srdce). 3) Krev neproudí plynule. 4) Podmínky výměny látek s tkáněmi jsou horší vzhledem k menší ploše styku hemolymfy s tkáněmi. 5) Transportní mechanizmus je sice méně energeticky náročný, je však také méně výkonný. 6) Omezená možnost regulace.
(s)
Dorsal vessel
Segment 13      Segment 12
(b)
50 cm H20
Gut       Septum   Ventral     Lateral      Nerve   ANTERIOR vessel      heart        cord
Blood pressure
Dorsal vessel
Ventral vessel
AAMViUVM
lllirllllllHlllrillllHllifiiliiiiJiiirliiiilNiilrriiliiiiiiiiiliiiiliHiLrnliiiJmifiiiiliiiil
Time (s)
figure f2-1     In the giant earthworm (Megascofides
australis), peristaltic contractions of the dorsal vessel ana1 pumping by the lateral hearts are both important in moving blood, (a) Blood flows from the dorsal vessel into the lateral hearts, present in the 1.1 anterior segments, and then is pumped into the ventral vessel, (b) Peak blood pressure in the dorsal vessef rs about twice as high as in the ventral vessef owing to peristaltic- contractions. (Adapted from Jones et al., 1994.1
(c?) The heart and the arteries emanating from the heart
Ostium     Anterior me^ian arte™ Dorsal abdominal artery
Hepatic artery
Anterior lateral artery
(fr) The array of suspensory ligaments around the heart
The heart is suspended within the array of suspensory ligaments in the pericardial sinus.
(c) Row of blood through the central circulation
Dorsal body wall of thorax
Suspensory ligament
Pericardial sinus
Bra n ch iopericardial "vein"
Oxygenated blood from the gills enters the pericardial sinus via
branchiopericardial "veins." Then.
.the oxygenated blood is sucked into the heart through the ostia during diastole and propelled into the arteries during systole.
Figure 24,22 The heart of a decapod crustacean    (a) The hear^
and the arteries leaving it. All the vessels connected to the heart 'n^t]c crustacean are arteries, (ŕ?) The heart is surrounded by an array o e ^ suspensory ligaments that run between the heart wall and the W3^^ the surrounding pericardial sinus, suspending the heart in the pe ^ dial sinus. (0 The position of the heart in the pericardial sinus^n,kens pattern of blood flow through the central circulation, [a after wi 1999; b after Plateau 1880,)
The circulator}' plan
I      | Oxygen poor J Oxygen rich
Systemic tissues
'(b) The hearts and gills of an octopus r vein
Efferent
branchial vessel
Afferent
branchial
vessel
The sysremic heart, branchial hearts, and gills are close neighbors anatomically.
fai Th e 24*20 Tne circulatory plan of squids and octopuses
one          angernent °ftne hearts, gills, and systemic tissues relative to
Pusean0ther' Hemocyanin'tne respiratory pigment of squids and octo-^ Sj tUrns blue when well oxygenated, but is clear or nearly clear deoxygenated. (6) A more realistic drawing of the central circula-
tory
system of an octopus. {6 after Johansen and Lenfant 1966.)
(a) Crayfish
Ostium
Efferent        Hemocoel      with valves       ^
branchial vesse
Arterial valve Artery
Afferent                Sternal
branchial vessel          sinus
(c) Cephaiopod
Eye
{b} Bivalve rnollusk
Cephas ic artery VentrfcFe
NepliridJal            Mantle
vessels             vessels
Vena cava          Afferent
cephafica    branchial vessel
Branchial heart
Sinus in foot
Hemocoel
Afferent branchial vesse
Ctenidial            Efferent
vessel      branchial vessel
Otevřené a uzavřené oběhové soustavy - srovnání.
TABLE 24.2 Systemic circulatory function in two decapod crustaceans and two fish of similar body sizes
Characteristics of circulatory function
Principal features of circulatory function
Rate of 02 delivery to tissues (mL 02/kg*vr\\n)b Rate of blood flow through systemic circuit (mL blood/kg*min)c Pressure change to perfuse systemic circuit (mm Hg)d Systemic resistance (pressure change divided by flow rate)e Secondary information Heart rate (beats7min) Stroke volume (mL/kg-stroke) Blood pressure in major systemic arteries (mm Hg)d Blood pressure in major systemic veins or venous sinuses (mm Hg)ú Blood oxygen-carrying capacity (vol %) Temperature during studies (°C) Body weight (g)
Spiny lobster (Panulirus interruptus)	Rock crab" {Cancer product us)	Starry flounder [Platkhthys stellatus)
0.80	0.60	0.46
128-148	125	39
14	3	16
0.1	0.03	0.4
65	101	35
2.1	1.2	1.2
35	10	18
21	7	2
2.0	1.3	5.7
16	12-16	8-11
515	-370	684
Rainbow trout trout
{Oncorynchus mykiss)
0.65 18 22 1.2
63 0.3
26 4
7.8 9-15 -210
ha\ Lacunar where blood flows most vigorously
(b) Muscle fibers in leg' coded red if highly C^-dependent
1 mm
The blue color marks well-perfused lacunar channels. Such channels tend to be most numerous and widest in the parts of the leg...
1 mm
.where the most 02-dependent muscle
fibers are found.
Figure 24.24 The microanatomy of blood flow in a lacunar system: Blood flows most vigorously where it is needed most
Adjacent cross sections of a tarantula's leg are shown, The tissue seen inside the cross sections is mostly muscle, that is, the tissue of the walking muscles, (a) Locations where blood flows particularly vigorously in the lacunar networks of the muscles, in this cross section, hemocyanin was stained irnmunohistochemically using monoclonal antibodies
(antibodies specific to hemocyanin). The stain was then visualized, [b] Locations of muscle fibers that are particularly C^-dependent, In this
cross section, fibers were assessed using their profiles of catabolic enzymes (similar to Figure 7.10). The fibers colored dark red are the most poised to produce ATP aerobically and are the most (^dependent Those colored light red are also aerobically poised, but less so. (From Paul et al. 1994J
o
Oběh hmyzu: septa, siny, pomocná srdce.
puisarne structures mat au legs, wings, and antennae. In blood of insects lacks a respir (From Jones 1964,)
Blood flows principally from posterior to anterior in the
dorsal part of the body...
Lateral a rte t v
...and from anterior to posterior in the ventral part of the body.
Křídlaté svaly-Nezbytné pro plnění u otevřených soustav. Spolupráce na rytmu.
Alary — 1 ■_:: _i.      .^—_^^___		
	/-j	W2
		
	* 1         ^"d	
	£g     gHy	
	B|Rffa ■ ■■■	
		■F^^^í
muscles       MW£		h?y^^   :■:
Pericardial       i ceils           (		
\	■ÜB-* v- "-riť^L ■ ■■ ■ í r^i~"Ui : ■ ■ -:-	
	[■' "BS^ i':::;vjj_.ii-^;-ri 11 fiíí^A"^. i V-' ■ ó" í?	
		MLÍár
		
Aorta
Heart
Segmental bfood vessel
Pomocná srdce na bázi nohou a řízení směru průtoku.
Coxa
Trochanter
Seplum
FIGURE   7.5.     An accessory pulsatile organ  in the  insect leg.  Arrows  show the hemolymph flow. From Hantschk (1991), Reprinted with permission.
Pomocná srdce na bázi tykadel.
Compound eye
Dilator muscle of ampulla
Accessory dilator^ muscle
Antennal vessel
Aorta
Esophagus
FIGURE 7.8.    An ampulla and its dilator muscles that pump hemolymph into the antennae. From Pass (1985). Reprinted with pennission.
Dýchací pigmenty - limit výkonu,
Prohemocyte
Plasmatocytes
Adipohemocyte
Spherule cell
Coagulocyte
^jjjjrjpr.
ranulocyte
Oenocytoid
Imunitní reakce.
TABLE 15-10							■		
Summary of	the development of immunity in			animals.	(Modified from Stites, Caldwell, und Pa\			-/a 1980,)	
		Immunologic				Non-	Phagocytic		
		Specificity	Immuno-			specific	Amehoid	Leukocyte	
	Graft	of Graft	logic	Phago-	Encapsu-	Humoral	Coelomo-	Diffcrcn*	Anti-
	Rejection	Rejection	Memory	cytosis	lation	Factors	cytes	tiation	bodies
Protozoans	Yes	No	No	Yes	No	No	No	No	No
j Poriferans	Yes	Yes	Yes	No	Yes	No	No	No	No
j Cnidaríans	Yes	Yes	Ye*	No	Yes	No	No	No	No
Annelids	Yes	Yes	Yľs	Yes	Yes	Yes	Yes	Probable	No
Mollasks	Yes	■>	•}	Yes	Yes	Yes	Yes	No	No
Arthropods	Yes	•j	>	Yes	Yes	Yes	Yes	No	No
Echinodcrms	Yes	Yes	Yes	Yes	Yes	Yes	Yes	Yes	No
Tunicates	Yes	Probable	Yes	Yes	Yes	Yes	Yes	Yes	No
Vertebrates	Yes	Yes	Yes	Yes	No	Yes	Yes	Yes	Yes
It was understood in the early 1930's that the immune response of insects is manifested both as cellular and humoral reactions.
... cellular reactions the microorganisms or the apoptotic cells are phagocytosed, ent              '              '   '
encapsulated by hemocytes.
In humoral defence processes three immediate reactions are triggered: melanisation, clotting of the hemolymph and synthesis of antimicrobial peptides.
;rs] p p 3 oBjMjTjijfi j J 9 lünuittJfiimfL
Invading microorganism
,|-     Cuticle
Pattern
recognition
proteins
Opsinizing proteins
Proteolytic cascades
0 I Epidermal cells
Epithelial antimicrobial response
Cellular defense by hemocytes
Systemic antimicrobial %^^^^f\   response by fat body
Fat body
FIGURE 7,17, Possible mechanisms of response to invading parasites by epidermal cells, hemocytes, and fat body. Epidermal cells produce antimicrobial compounds, and pattern recognition and opsinizing proteins target invaders for attack by hemocytes. Fat body cells can also mount a systemic antimicrobial response.
Figure 5: Diagram of an insect granular cell emphasizing its multifunctional role.
LYSOZYME
(Lysis of bacteria)
PHAGOCYTOSIS
COAGULOGENS
(sealing of wounds)
STICKY PROTEINS
(opsonisation and
recognition of foreign particles)
AGGLUTININS
(coating of foreign agents)
CHEMOTACTIC FACTORS?
(attraction of plasrnatocytes during
nodule formation
/e nc ap sul ati o n/ wo und
healing)
MELANIN
(killing of parasites?)
Nodule formation
During nodule formation insect hemocytes aggregate to entra bacteria. Nodules can attach to tissues or may be encapsulated.
An insect lectin scolexin was found to be involved in nodul formation in Manduca sexta. Scolexin is produced by epidermal and midgut cells upon wounding or bacterial infection.
In the medfly (Ceratitis capitata), a protein with molecular mass o. 47  kDa  is secreted  by  hemocytes after LPS stimulation  and
nee of tvrosine and tvrosinase.
Figure 10: Nodule formation in fatbody and trachea of B.mori. Magnification 60X
Figure 11: Mature dark melanized nodule of B.mori as observed under phase contrast microscope at 600X magnification
The formation of the black pigment, melanin is catalysed by the enzyme phenoloxidase, which is converted to its active form by a serine protease cascade.
The inactive proenzyme, prophenoloxidase is synthesized in the hemocytes and after releasing by cell rupture it is either actively transported to the cuticle or deposite " around wounds and encapsulated parasites.
Prophenoloxidase has been purified and subsequently characterized from the hemolymph of a range of insect species.
insect prophenoloxidase enzyme contains a sequence with similarity to the thiol-ester region of the vertebrate comolement component oroteins C3 and C4.
Enkapsulace a nodulace. Fenoloxidázová kaskáda.
CW.CHCOOH
NWu
HO..   v^v^-C-MiCM COOH
$«J*oloKv>te2A.     VV5ÍSv'^>
l*2ľ*l

ACETYL OöRtoiAi
O
'/
o        o
/^R.+ HA*?rB"
(Invading microorganism
Pattern
recognition
proteins
Serine protease cascade
Melanin
Serine protease inhibitor
Prophenoloxidase
------►
i
Phenoloxidase
Phenols
i
+ Quinones
A/-acetyldopamine
Crystal cell
Tyrosine
FIGURE 7,18,    Mechanism of humoral encapsulation by the deposition of melanin on foreign invaders. The serine protease inhibitor restricts phenoloxidase activity to the site of the infection.
Figure 16: <B(6odcCotting system (horseshoe crab)
Horseshoe crab blood clotting system can be activated by bacterial LPS or fugal beta-1,3-glucans. Binding of LPS to Factor C auto activates Factor C, which then triggers the serine proteinase cascade, leading to activation of proclotting enzyme. Clotting enzyme cleaves coagulogen to form gel-like materials coagulin. Another pathway of this system is activated by beta-1,3-glucans, in which binding of glucans to Factor G auto activates Factor G.
rote i n
The third humoral reaction to infection is the rapid de novo synthesis of a battery of antimicrobial peptides (Boman, 1996; Hetruetal., 1998).
The principal site of synthesis is the fat body, but also the hemocytes, the cuticular epithelial cells, the gut, the salivary gland, and also the reproductive tract.
During the research work conducted in the last decade nearly 60 peptide antibiotics have been described in insects. Though diverse in structure, they are basically amphipathic molecules acting at membranes and thereby killing the target cell eventually by lysis.
The insect antimicrobial proteins are grouped into families, based on structural and sequence similarities and their proposed targe" in the bacterial cell wall.
The attacin-like bacteria inducible proteins have been identified in butterflies and in Drosophila. These proteins are active only against gram negative organisms, where they affect cell division mechanisms by inhibiting the synthesis of outer membrane proteins. The family of these factors includes glycine-rich peptides (20-28kDa) characterized by the presence of one or more
ni.sRTOi
Lysozyme hydrolyses ß-(1,4)-glycosidic bonds in peptidoglycan of bacterial cell wall. Insect lysozymes are proteins(14 kDa) with sequence similarity to vertebrate lysozymes. The lysozyme gene or cDNAs have been cloned in several insect species (Kylsten et al., 1992; Sun et al., 1991; Lee & Brey, 1995). In Drosophila, the lysozymes are encoded by at least seven genes and expressed in different parts of the digestive tract and at different stages of development.
Cecropins have antibacterial activity against both gram-positive and gram negative bacteria since they interact with lipid membranes forming voltage dependent ion channels. The cecropins (4 kDa) are devoid of cystein, and exhibit a structure of two a-helices joined by a hinge region. Families of cecropin genes with some sequence differences have been found in butterfly species, in the flesh fly (Sarcophaga peregrina) and in Drosophila. A mammalian cecropin was identified in pig intestine (Lee et al., 1996) and bovine adrenal glands (Strub et al., 1995) which implies that cecropins may be widespread in the animal kingdom.
Defensins attack mainly Gram-positive bacteria in contrast to attacin-like antibacterial peptides. They act on the cytoplasmic membrane and lyse cells by the formation of membrane channels. Insect defensins are cationic peptides (4 kDa) containing six conserved cystein residues engaged in three disulphide bridges. They possess three distinct domains: amino-terminal loop, an ct-helix, and an antiparallel ß sheet. About thirty defensins have been characterized in various insect species. Although numerous
e proline rich antimicrobial peptides lyse Gram-negative bacteria by increasing the membrane permeability. They are peptides with molecular mass of 2 - 4 kDa, lacking cysteine and containing at least 25% proline. The O-glycosylation at the threonin residues is essential for their biological activity. Apaedicins and abaecin from honey-bee, drosocin and metchnikowin from Drosophila, pyrrhocoricin, lebocin and metalnikowin belong to this family. Also, the pig intestine and bovine leukocytes have been shown to produce proline-rich antibacterial peptides, although these peptides do not share sequence homology to the proline-rich peptides of insects.
Diptericins have so far been described only in dipteran species. They are 9-kDa peptides containing both an attacin-like G domain, a C-terminal glycine rich residue and a short N-terminal proline-rich   region  containing  a  consensus  site for  O-glycosylation.
Diptericins are lytic for Gram-negative bacteri—,:~'--------'-----
to a way of action similar to that of attacin
Other inducible antibacterial proteins have been isolated fro insects not fitting into the groups described above. Coleptericin, holotricin-2, hemiptericin and gallysin-1  act on Gram negative
iay b
o fp\/1 n i n p      ■Fill
frog-s
.—x-„:_    while   moricin,   thanatin   (homolog—   A~   *—
the     brevinir
hymenoptaecin can lyse both Gram-negative and Gram-pos bacteria.
[UM
Recently inducible peptide antibiotics against fungi have been discovered in insects. The peptide named AFP, tenecin-3 and holotricin-3 share similarities while drosomycin shows a significant homology with a family of plant antifungal peptides, lermore, the antibacterial peptides metchnikowin and nanatin have antifungal activity.
Parallels between humoral immune response of insects and mammalian acute phase response:
•The acute phase response of mammals is stimulated by tissue injury or bacterial challenge and characterized by the immediate synthesis of acute phase proteins mainly in the liver and in cells of the innate immune system. The synthesized proteins act as opsonins, blood clotting and wound healing factors. In insects, the rapid humoral immune reactions are also triggered by wound or microbial infection. They involve proteolytic cascades leading to hemolymph clotting and the rapid de novo synthesis of antimicrobial peptides by the fat body, the functional analogue of the mammalian liver, and the hemocytes.