Download Heart histology with four chambers in the spotted scat, Scatophagus

Heart histology with four chambers in the spotted scat,
Scatophagus argus during juvenile stage
1
2
3
4
Running head Heart histology with four chambers in the spotted scat,
5
Scatophagus argus
6
7
Lamai Thonhboon1, Sinlapachai Senarat2,*, Jes Kettratad2,
8
Mark Tunmore3, Pisit Poolprasert4, Sansareeya Wangkulangkul1,
9
Watiporn Yenchum5, Wannee Jiraungkoorskul6
10
1
11
Department of Biology, Faculty of Science, Prince of Songkla University, Songkhla,
12
13
90110, Thailand
2
Department of Marine Science, Faculty of Science, Chulalongkorn University, Bangkok,
14
10330, Thailand
3
15
4
16
Program of Biology, Faculty of Science and Technology, Pibulsongkram Rajabhat
17
University, Phitsanulok, 65000, Thailand
5
18
19
20
21
The Boat House, Church Cove, Lizard, Cornwall, TR12 7PH England
Bio-Analysis Laboratory, Department of Chemical Metrology and Biometry,
National Institute of Metrology (Thailand), Pathum Thani, 10120, Thailand
6
Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, 10400,
Thailand
22
23
24
*Corresponding author, e-mail: [email protected]
25
Abstract
26
The heart structure of spotted scat, Scatophagus argus was observed from the
27
Paknam Pranburi Estuary, Thailand, using histological and histochemical
28
techniques. The results revealed that the heart structure consisted of four
29
chambers including the sinus venosus, the atrium, the ventricle and the
30
bulbus arteriosus, which were basically structured to three layers (the
31
epicardium, the myocardium and the endocardium). The feature of the sinus
32
venosus was similarly structured to the atrium. It had two layers including a
33
thin layer of the epicardium and the endocardium. However, the
34
myocardium in the atrium had much more cardiac muscular tissue than
35
sinus venosus. The ventricle in this species was thicker than that of the
36
atrium and composed of the thickest layer of the cardiomyocytes. The
37
localization of the bulbus arteriosus with the ventral aorta was finally
38
observed of the heart organ, which had a thicker layer of the myocardium
39
than other layers.
40
41
Keywords: Blood vessel, Fish, Heart, Histology, Spotted scat
42
43
Introduction
44
Under functional structures, the cardiovascular system (including the
45
heart, blood vessels and blood components) of a number of teleost species are an
46
important role in the blood pressure, the chemical components and transporting of
47
nutrient and oxygen (Olson, 2000, Roberts, 2012). Therefore the anatomy and
48
histology of this system has been studied in Perca fluviatis (Pollak, 1960),
49
Prochilodus lineatus and Hoplias malabaricus (Rivaroli et al., 2006), which
50
provided not only structural information but also applied to further studies, for
51
example histopathology and physiology. However, there are no investigations of
52
the histology and histochemistry in the Scatophagus argus during the juvenile
53
stage. It is an important mangrove species and dominantly found in the Paknam
54
Pranburi Estuary, Thailand. Additionally, this species is considered to be very
55
important, both commercially and in the food web/food chain.
56
57
Materials and methods
58
Ten specimens of Scatophagus argus with total length about 3.4±0.21 cm
59
were collected from 2 stations (N 12°24'15.8" / E 099°58'25.6" 2 and N
60
12°24'21.6" / E 099°58'37.1") in the Paknam Pranburi Estuary, Thailand. All fish
61
were euthanized by a rapid cooling shock (Wilson et al., 2009). They were then
62
fixed in Davidson's fixative (about 24 - 36 h) and transferred to 70% ethanol. The
63
heart of this fish were collected and subsequently subjected to standard
64
histological techniques (Presnell and Schreibman, 1997; Suvarna et al., 2013).
65
Sections at 5 - 6 µm thickness were subsequently stained with Harris's
66
haematoxylin and eosin (H&E), Alcian blue (AB) and periodic acid-Schiff (PAS)
67
(Presnell and Schreibman, 1997; Suvarna et al., 2013).
68
69
Results and Discussion
70
Morphology and histology of the heart
71
Morphological observation of the heart in S. argus was anteriorly located
72
in the peritoneal cavity and caudo-ventrally to the gill. It was likely investigated in
73
some teleosts (Poppe and Ferguson, 2006, Senarat et al., 2016). Under light
74
microscope, the heart structure revealed that it typically consisted of four
75
chambers including the sinus venosus (data not shown), the soft atrium, the
76
muscular ventricle and the bulbus arteriosus (Figs. 1A-1E). Similarly, the heart
77
morphology in Rastrelliger brachysoma (Senarat et al., 2016) and Poecilia
78
reticulata (Genten et al., 2008) was studied.
79
Based on histology and histochemistry, the heart wall was normally
80
composed of three layers such as the outermost epicardium, middle myocardium
81
and innermost endocardium in longitudinal sections (Figs. 1B-1C). The outermost
82
epicardium originated from the pericardial sac. A single sub-layer of flattened
83
epithelial cells tightly packed in this layer, however, a prominent loss connective
84
tissue was observed. The middle layer was composed of the thickness of the
85
middle myocardium. Within this layer, the specialized cardiac muscle was mainly
86
seen around the heart wall. Each cardiac muscle contained a great syncytium of
87
anastomosing fiber (cardiomyocytes). As a result, the function of cardiomyocytes
88
is an important process in the bind natriuretic peptides (Cerra et al., 1997) and the
89
anti-freeze mucins (Icardo, 2012). The innermost endocardium showed that a thin
90
layer of loose connective tissue and epithelial cells was observed.
91
The structure of the sinus venosus was likely seen in the general structure. A thin
92
sinus venosus wall was divided into two layers (a thin layer of epicardium and
93
endocardium) (data not shown). However, the myocardium layer was markedly
94
found, as previously seen in R. brachysoma (Senarat et al., 2016). On the other
95
hand, the sinus venosus in Anguilla anguilla and Pleuronectes platessa was found
96
(Santer and Cobb, 1972; Yamauchi, 1980). The function of this structure has been
97
reported to concern storages of the incoming venous blood from the hepatic portal
98
vein (Genten et al., 2008). The wall of the atrium was thin-walled and
99
histologically similar in the sinus venosus (Figs. 1A-1C). However, some
100
characterization of this chamber was different, that is, the myocardium had much
101
more cardiac muscular tissue than sinus venosus. The cardiac muscular tissue
102
reacted positively with PAS (Fig. 1C), indicating the presence of glycoprotein.
103
The atrium is known to play an important role in supporting the atrial architecture
104
and blood pump (Icardo, 2012). The blood from the fish body enters the atrium
105
via the sinus venosus. The atrium contains the cells that initiate the contractions
106
for pumping the blood into the ventricle (Icardo, 2012). It was known that the
107
ventricle chamber was thicker than that the atrium (Fig. 1B). Interestingly, the
108
thickest layer of cardiomyocytes was observed in the ventricular myocardium,
109
which reacted positively with PAS reaction (Fig. 1C). This agreed with previous
110
reports in Echiicthys vipera, Oncorhynchus mikiss and Salvelinus alpinus (Icardo,
111
2012). It is possible that the roles of the structure was concerned with supporting
112
the pumping action in teleost (Genten et al., 2008, Roberts, 2012). Additionally in
113
our study, two layers, including an outer compact and an inner layer of the
114
abundant spongy layers in the ventricle was also observed, similar to that of
115
Danio rerio (Menke et al., 2011). The bulbus arteriosus was a lastly region of the
116
heart. The wall of this chamber histologically revealed that the epicardium and the
117
endothelium was a thin layer, in contrast, the myocardium was a thick layer (Figs.
118
1A, 1D) and reacted positively with AB (Fig. 1E). The functional structure of the
119
bulbus arteriosus is to regulate the pressure of the blood leaving the heart (Genten
120
et al., 2008).
121
122
Histology and histochemistry of the aortas
123
The blood vessels in both dorsal and ventral aortas were obviously seen in
124
S. argus (Fig. 2). Histological localization of these vessels was formed, which
125
ventral aorta was jointed with the bulbus arteriosus and ran the mandibular region
126
via the afferent bronchial arteries. The sinus venosus was connected by dorsal
127
aorta. The dorsal aorta had similarly structure that could be classified into three
128
layers, including tunica intima, tunica media and tunica adventitia (Figs. 2A-2C).
129
This was supported by higher vertebrates (Genten et al., 2008). In the tunica
130
intima, its epithelium was lined by two sub-layers (endothelial epithelium and
131
subendothelial sub-layers). The internal elastic layer was not observed.
132
tunica media lay between the tunica intima on the inside and the tunica adventitia
133
on the outside. The prominent elastic, reticular fibers and a few smooth muscles
134
was the majority layer in the tunica media. It is possible that this characterization
135
has important roles in the constriction and dilation of the vessel (Groman, 1982;
136
Roberts, 2012). Lastly, the outer layer of the tunica adventitia was covered by a
137
thin connective tissue and a few collagen fibers. The adventitia function is a
138
fibroblast and stem cell progenitor reservoir and allows the rapid influx of these
139
cells into the aortic wall (Psaltis et al., 2011).
The
140
Conclusions
141
The present study showed that the heart structure of S. argus was typically
142
composed of four chambers, including the sinus venosus, the soft atrium, the
143
muscular ventricle and the bulbus arteriosus, which was newly reported. Our
144
histological results will be applied to further study for example ultrastructure,
145
histopathology and physiology.
146
147
Acknowledgement
148
We thank the members of the Microtechniquce and Histology laboratory,
149
Department of Biology, Faculty of Science, Prince of Songkla University,
150
Thailand for their technical support in laboratory.
151
152
References
153
Cerra, M.C., Canonaco, M., Acierno, R. and B. Tota. (1997). Different binding
154
activities of A- and B-type natriuretic hormones in the heart of two Antarctic
155
teleosts, the red-blooded Trematomus bernacchii and the hemoglobinless
156
Chionodraco hamatus. Comp. Biochem. Physiol., 118A: 993–999.
157
158
159
160
Genten, F., Terwinghe, E. and A. Danguy. (2008). Atlas of Fish Histology.
Enfield, Science Publishers, 219 p.
Groman, D.B. (1982). Histology of the Striped Bass. USA, American Fisheries
Society, 129 p.
161
Icardo, J.M. (2012). The Teleost Heart: A Morphological Approach. In: Ontogeny
162
and Phylogeny of the Vertebrate Heart. Sedmera, D., and Wang, T., (eds).
163
Springer Science, Business Media, LLC, p. 35-53.
164
Menke, A.L., Spitsbergen, J.M., Wolterbeek, A.P. and Woutersen, R.A. (2011).
165
Normal anatomy and histology of the adult zebrafish. Toxicol. Pathol., 39:
166
759-775.
167
168
Olson, K.R. (2000). Circulation System. In: The Laboratory Fish. Ostrander,
G.K., (eds). 2nd ed, UK, Academic Press, p. 369-378.
169
Pollak, A. (1960). The main vessels of the body and the muscles in some teleost
170
fish. Part I. The perch (Perca fluviatilis L.). Acta Biologica Cracoviensia
171
Series Zoologica, 3: 115–138.
172
Poppe, T.T. and Ferguson, H.W. (2006). Cardiovascular System. In: Systemic
173
Pathology of Fish. Ferguson H.W., (eds). 2nd ed,. UK, London, Scotian Press,
174
p. 141-167.
175
176
Presnell, J.K. and Schreibman, M.P. (1997). Humason’s Animal Tissue
Techniques. 5th ed. US, Johns Hopkins University Press, 600 p.
177
Psaltis, P.J., Harbuzariu, A., Delacroix, S., Holroyd, E.W. and Simari, R.D.
178
(2011). Resident vascular progenitor cells – diverse origins, phenotype,
179
and function. J. Cardiovasc. Transl. Res., 4: 161 – 176.
180
Rivaroli, L., Rantin, F.T. and A.L. Kalinin. (2006). Cardiac function of two
181
ecologically distinct Neotropical freshwater fish: Curimbata, Prochilodus
182
lineatus (Teleostei, Prochilodontidae), and trahira, Hoplias malabaricus
183
(Teleostei, Erythrinidae). Comp. Biochem. Physiol., Part A 145: 322-327.
184
Roberts, J.R. (2012). Fish Pathology. 4th ed. USA, Wiley. 508 p.
185
Santer, R.M. and Cobb. J.L.S. (1972). The fine structure of the heart of the teleost,
186
Pleuronectes platessa L. Zeitschrift fur Zellforschung und mikroskopische
187
Anatomie, 131: 1–14.
188
Senarat, S., Kettretad, J. and Jiraungkoorskul, W. 2016. Histological evidence of
189
the heart and ovarian vessels in the short mackerel, Rastrelliger brachysoma.
190
EurAsian J. BioSci., 10: 13-21.
191
192
Suvarna, K.S., Layton, C. and Bancroft. J.D. (2013). Bancroft’s Theory and
Practice of Histological Techniques. 7th ed. Canada, Elsevier, 654 p.
193
Wilson, J.M., Bunte, R.M. and A.J. Carty. (2009). Evaluation of rapid cooling and
194
tricaine methanesulfonate (MS 222) as methods of euthanasia in zebrafish
195
(Danio rerio). J. Am. Assoc. Lab. Anim. Sci. 48: 785-789.
196
197
198
199
200
201
202
203
Yamauchi, A. (1980). Fine Structure of the Fish Heart. In: Heart and heart-like
organs. Bourne G., (eds).. US, New York, Academic, p. 119–148
204
Figure legends:
205
Figure 1. Anatomy and light micrograph of the heart in Scatophagus argus;
206
200 μm (A), 20 μm (B-D). At = atrium, Ba = bulbus arteriosus, Ed =
207
endocardium, Epi = epicardium, Ery = erythrocytes, My = myocardium, V =
208
ventricle, Va = ventral aorta. Harris's haematoxylin and eosin (A, B, D),
209
periodic acid-Schiff (C) and Alcian blue (E).
210
Figure 2. Light micrograph and schematic diagram of ventral aorta (A-B)
211
and dorsal aorta (C) in Scatophagus argus; 30 μm (A-C). Da = dorsal aorta,
212
Va = ventral aorta. Harris's haematoxylin and eosin (A, C), periodic acid-
213
Schiff (B).
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
Figure 1
229
230
231
Figure. 2