J. Zool., Lond.

( A ) (1986) 209,305-317

Scanning electron microscopy of the accessory respiratory organs of the

Snake-headed fish, Channa striata (Bloch) (Channidae, Channiformes)
G . M . HUGHESA N D J . S . D . MUNSHI*
Research Unit for Comparative Animal Respiration, Bristol University, Woodland Road, Bristol,
BS8 1UG

(Accepted 27 August 1985)

(With 4 plates and 5 figures in the text)

The accessory organs of this swamp-living fish have been studied using SEM and light
microscopy.
Accessory organs are found in the suprabranchial chambers, labyrinthine plates on the first
epibranchial, and roof of the buccopharynx. In these regions, there are nodules of respiratory
islets each consisting of many vascular papillae. Non-respiratory ‘lanes’ separate the islets and
are covered with microridged epithelial cells. The dome-shaped papillae have a smooth surface
with band-like structures running over them. Microvilli are present only at the base of the
papillae. The vascular papillae of the buccopharynx are lodged in cup-like receptacles into
which they can retract.
The intraepithelial capillaries contain unicellular valves which project into the lumen of the
papillae.
Capillaries supplying the papillae have an undulatory path which represents a second stage
in the evolution of this type of structure.

Contents
Page
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305
Materials and methods ........ . . . . . . . . . . . . . . ....... 307
Observations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 312
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 316
Abbreviations used on figures and plates ..................... 317

Introduction
Channa (= Ophicephalus) striata lives in freshwater swamps, pools and chaurs**, where the
macrovegetation is abundant and the water hypoxic. These swamps are characterized by low-
lying, water-logged shallow areas with a loose peaty bottom rich in decaying organic matter.
The fish is highly predatory, feeding on fishes, frogs, insects and other live food. In the daytime,
they hide within the cover of floating macrovegetation and during the night are found near the
shallow areas of ponds and swamps. Dehadrai (1962) stated that Channa striata could live out
of water for more than 28 hours. It moves on land by means of the tail and pectoral fins. Its
* Present address: Postgraduate Department of Zoology, Bhagalpur University, Bhagalpur, India
** A wetland area with extensive aquatic vegetation
305
0022-5460/86/007305+ 13 $0340 @ 1986 The Zoological Society of London
306

3rd and 4th LAB

rd
G. M. H U G H E S A N D J. S. D. M U N S H I

“BFt LAB

EB

FIG.I . (a) Lateral view of dissection of the head of Channa srriara, showing the extension of suprabranchial chamber,
labyrinthine organ, and the levator muscles of the gill arches. (b) First gill arch showing the labyrinthine organ and gill
filaments.
 ACCESSORY RESPIRATORY O R G A N S OF SNAKE-HEADED FISH 307
air-breathing habits, from the very early larval stages, has attracted the attention of many zoolo-
gists in the past including Day (1868), Dobson (l874), Ghosh (1934) and Hora (1935). Munshi
(1960, 1962b) and Hughes & Munshi (1973b) studied the structure of the respiratory organs of
this fish using transmission electron microscopy. Evidence was obtained supporting the view that
the air-breathing organs do not represent modified gills in this species.
As a scanning electron microscopic study of accessory respiratory organs in the catfish Heterop-
neustes (Hughes & Munshi, 1978) gave invaluable support to the homology of these structures
with gill primary and secondary lamellae, it was decided to extend our previous investigations to
another Indian air-breathing fish, namely Channa. SEM is particularly suited to investigations
of the gross surface morphology, but also the detailed nature of the gas exchange surfaces. Light
microscopy was used to supplement both the present and previous studies.

Materials and methods

Specimens of Channa striatu were collected from the swamps of North Bihar. The specimens were
anaesthetized in MS222 and the accessory respiratory organs rapidly dissected and fixed in different
glutaraldehyde solutions (2.5% and 2573, using a phosphate/cacodylate buffer at pH 7.4.Following fixation
overnight, this material was post-fixed in osmium tetroxide (1% in cacodylate buffer) before critical point
drying and gold coating. Observations were made with a Cambridge S4 scanning electron microscope.
Some of the material was embedded in araldite, sectioned (61 pm) with an LKB Ultratome and stained
in toluidine blue for light microscopy.

Observations
In C. striata, the suprabranchial chambers develop as pharyngeal pouches dorsal to the gill
arches, and lateral to the auditory capsule (Fig. la). The suprabranchial chamber is incompletely
divided into two compartments by a shelf-like transverse cartilaginous/bony outgrowth from the
hyomandibula (Fig. 2). The pharynx opens into the anterior compartment through an unguarded
inhalent aperture. The posterior compartment is larger in size, extending backwards as a deep
recess. It also communicates independently with the pharynx, through an opening which is
guarded by a ‘shutter’ or labyrinthine plate borne by the epibranchial bone of the first branchial
arch (Figs lb, 2). The dorsal part of the first gill slit forms the exhalent aperture for the
suprabranchial chamber. But as the anterior inhalent apertures are not provided with shutter-
like structures, the water or air may enter and leave the suprabranchial chambers through these
apertures during either aquatic or aerial ventilation.
The roof of the buccopharynx is covered with vascular respiratory islets where gas exchange
occurs (Fig. 2).
In C. striata, nodular structures develop on the margins of the epibranchial plate giving it a
labyrinthine appearance (Fig. 1 b, Plate IIIa). These structures are probably involved in both
aquatic and aerial respiration.
The epithelia lining the suprabranchial chambers, labyrinthine organs and the nodules are all
highly vascularized. The vascularized epithelium extends into the roof of the buccopharynx. The
epithelium of the anterior part of the pharynx and the hind part of the buccal cavity covers
numerous vascular patches, the respiratory islets. Each islet may have hundreds of vascular
papillae which form bunches of grape-like structures projecting above the general surface of the
epithelium (Plates I, 11). Between the vascular respiratory islets are found non-respiratory areas
in the form of ‘lanes’ (Plates Ic, IIb).
308 G . M. H U G H E S A N D J . S . D. M U N S H I

RI

-IM

1st GA

12nd GA

OP - 3rd GA

Oes
FIG.2. Ventral dissection of the head showing the respiratory islets on the dorsal surface of buccopharynx, the
inhalent apertures, labyrinthine organ of the hyomandibula, shutter-like structure of the first gill arch and the
oesophagus.

PLATEI. SEM of the buccopharynx of Chunnu sfriutu. (a) Enlarged view of vascular papillae of buccopharynx,
one of which is retracted within the receptacle. Prominent, band-like epithelial cell boundaries are seen on the surface of
the vascular papillae. Prominent microridged epithelial cells are found in the non-vascular area. Microvilli are visible at
the base between the vascular papillae ( x 7100). (b) Higher magnification showing the alternate arrangements of
vascular papillae in different stages of retraction. The prominent rims of the receptacles are seen as band-like structures
connected strongly with each other ( x 7350). (c) View of the vascular and non-vascular areas of buccopharynx showing
whorls of microridges on the epithelial cell surface. In the transitional area, the microridges break up into microvilli
( x 2725). (d) Fully protracted vascular papillae showing the loop-like formations of blood vessels. Prominent band-like
cell boundaries of the epithelial cells and the alternate arrangement of vascular papillae are clearly discernible ( x 7 100).
ACCESSORY RESPIRATORY O R G A N S OF S N A K E - H E A D E D FISH 309
 PLATE11. SEM of surface of respiratory air-sac (=suprabranchial chamber) of C. siriuru. (a) A bunch of fully
protracted vascular papillae of air-sac without receptacles and bands ( x 3400).(b) Higher magnification of vascular and
non-vascular areas of respiratory air-sac showing vascular papillae with microvilli at their bases and microridge bearing
epithelial cells in the non-respiratory area. The microridges become irregular and then break up into microvilli in the
transitional area ( x 3300).
 PLATE111. SEM of labyrinthine organ of Chunna striara. (a) Dorso-lateral view of part of the labyrinthine organ
showing a few crenulated plates ( x 22). (b) View of vascular and non-vascular areas of labyrinthine organ, showing
the arrangement of vascular papillae and microridged epithelial cells of non-vascular area of the labyrinthine organ
( x 3700). (c) Higher magnification of labyrinthine surface showing a retracted vascular papilla within the cuplike
receptacle. The inner smooth surface and the prominent rim of the receptacle is seen. The irregular microridges break
up into microvilli in the transitional region ( x 6450).
312 G. M . H U G H E S A N D J . S. D. M U N S H I

The vascular papillae of the labyrinthine organ are dome-shaped structures which bulge out
from the surface and are in close contact with each other (Plate III(b)). The outer epithelial
surface of the papillae is smooth with band-like structures running over it which may denote the
cell boundaries. Microvilli are present only at the bases of papillae (Plate IIIb, c). The non-
vascular areas are lined by characteristic microridged epithelial cells (Plate IIIb). Plate IIIc is a
transitional area showing cells bearing microvilli as well as microridged cells.
The vascular papillae of the buccopharynx are of a specialized nature, being lodged in cup-
like receptacles (Plate Ia-c), the edges of which are connected with each other by thick bands as
shown in Plate Ib. Some of the papillae are seen retracted inside the cup-like depressions (Plate
Ia-c). Plate IIIc is a remarkable micrograph showing a retracted papilla and the inner surface of
the receptacle. Coagulated mucus is also visible in various places on the surface of non-vascular
areas.

Light microscopy
A vertical section of a respiratory islet, under high magnification, shows blood capillaries
forming vascular papillae jutting out on the surface of the epithelium (Plate IVa-c). Each sub-
epithelial arteriole comes to the surface of the suprabranchial chamber at several points so that
it has a wave-like shape (Figs 4, 3a, Plate IVd). Epithelial cells separate the vascular papillae
from each other. The lining of the papilla wall is formed of endothelial cells, the nuclei of which
lie at the base. The projecting part of the papilla is dome-shaped and its wall is extremely thin.
Unicellular valves are invariably found projecting inside the lumen of the vascular papillae. The
valves presumably influence the movement of erythrocytes and other blood cells through the
vascular papillae as they pass from one recess to the other part of the papillae (Plate IVd).
The intraepithelial arterioles traverse the epithelium, having a wave-like or spiral organization,
jutting out at places to form the vascular papillae before opening into veins (Fig. 3a, b).

Discussion
The respiratory islets of C. striata are composed of hundreds of vascular papillae which are
specialized parts of intraepithelial arterioles. The respiratory islets are supplied with mixed blood
from labyrinthine arteries that originate from the first and second pairs of afferent branchial
arteries. Oxygenated blood is drained from the suprabranchial chamber by suprabranchial veins
which lead into the jugular veins (Das & Saxena, 1956; Munshi, 1983). Satchel1 (1976) gives a
useful comparison of the blood supply in Channa with that of other air-breathing fishes.
The vascular papillae of the buccopharynx are more specialized than those of the suprabranchial
chamber and labyrinthine plates as they are retractile. There appears to be some mechanism to
shunt blood away from the respiratory islets of this region, presumably when the fish manipulates
food in its buccal cavity. As a result, the vascular papillae are retracted back into the

PLATEIV. Photomicrographsof vertical sections of air-sacof C. srriara. (a) Vertical section of wall of air-sac showing
vascular respiratory epithelium, vascular papillae and dermis ( x 1400). (b) Vertical section of air-sac showing structure
of the vascular papillae with their characteristic septa-like valves, RBC's, epithelial cells and connective tissue cells
( x 1400). (c) Tangential section of vascular papillae showing valves separating their afferent and efferent pathways,
cuboidal and stratified epithelial cells and basophilic mast cells ( x 1400). (d) Vertical section of air-sac epithelium
showing the wave-like configuration of intraepithelial capillary to form vascular papillae with septa-like valves, epithelial
cells and connective tissue ( x 1400).
ACCESSORY RESPIRATORY ORGANS O F SNAKE-HEADED FISH 313
314 G . M . H U G H E S A N D J . S. D . M U N S H I

FIG.3. (a) Diagrammatic representation showing the wave-like arrangement of blood capillaries to form vascular
papillae of a respiratory islet of C. sfriara. (b) The rims of the receptacles of vascular papillae and microvillous and
microridged epithelial cells are also shown.

CPC VP

FIG.4. Diagram of a vertical section through a respiratory islet to show the position of the valve in each vascular
papilla and the ‘cap cells’ of the rims of the receptacles.
 ACCESSORY RESPIRATORY ORGANS O F SNAKE-HEADED FISH 315

BM

FIG. 5. (a) Diagrammatic view showing the ultrastructure of a vascular papilla of C. striafa showing position of the
valve, cap cells and the afferent and the efferent pathways of blood flow. (b) Diagram based on the ultrastructure of one
vascular papilla cut at the base, with afferent and efferent pathways of blood separated by a septum-like valve composed
of connective tissue and endothelial lining which under light microscope looks like a pillar cell with columns.

cup-like depressions of the buccopharyngealepithelium. There are prominent band-like structures

on the epithelial surface which connect one papilla with another. The epithelial cap-like structures
seen on the surface of vascular papillae under TEM by Hughes & Munshi (19736) may have
been sections through the rims of cups of such papillae observed under SEM in the present study.

Structural evolution of vascular papillae

The vascular papillae are sections of capillaries which connect the arterial system of the
branchial region with those of the veins of the jugular system and which are specialized for gas
exchange. Similar types of vascular papillae are also found in the respiratory islets of Monopterus
cuchia, an air-breathing fish belonging to the order Symbranchiformes (Munshi & Singh, 1968).
In both Channa and Monopterus, the respiratory islets are distributed not only in the respiratory
sacs but also in the buccopharynx. However, in M. cuchia, where the gills are very much reduced,
the second branchial arch only bears a few gill filaments; the respiratory islets have de-
veloped extensively in the pharynx as well as in the hypopharynx (Munshi er al., subm.). In the
climbing perch, Anabas testudineus (Perciformes), the respiratory islets are restricted to the
labyrinthine organs and epithelial lining of the suprabranchial chambers (Hughes & Munshi,
1973~).In this fish, the respiratory islets are composed of a series of parallel blood capillaries,
316 G . M . H U G H E S A N D J . S. D . M U N S H I

each capillary being lined by a single row of endothelial cells (Hughes & Munshi, 1973~).Their
prominent cell bodies project into the capillary lumen. The endothelial cells have tongue-like
processes which act as minute valves controlling the flow of blood. These parallel capillaries are
separated from each other, not by pillar cells but by supporting epithelial cells. This Anabus
model seems to represent an early stage in the evolution of air-breathing organs of these groups
of fishes in which arterio-venous connections form straight endothelial tubes situated just below
the epithelium. At a later stage, they became arranged in an undulatory fashion as in Chunna
(Fig. 3a). The valves here are elongated structures having a core of basement membrane and
lined by flat endothelial cells. The functional significance of this wavelspiral-like arrangement of
capillaries giving rise to a series of vascular papillae seems to be that a greater volume of blood
could be accommodated for a given surface of respiratory islets. Moreover, the system provides
an efficient mechanism for bringing the erythrocytes into contact with the gas exchange surface.
The greater capillary length would lead to a longer contact time for equilibration of the blood
and respiratory medium. It also increases the resistance to blood flow.

We thank the University Grants Commission, New Delhi and University of Bristol for providing financial
assistance.

REFERENCES
Das, S. M. & Saxena, D. B. (1956). Circulation of the blood in the respiratory region of the fishes Labeo rohita and
Ophicephalus strialus. Copeia 1956 100- 109.
Day, F. (1868). Observations on some of the freshwater fishes of India. Proc. zool. SOC.Lond. 1868: 274-288.
Dehadrai, P. V. (1962). Observations on certain physiological reactions in Ophicephalus striatus exposed to air. Lye Sci.
11: 653-657.
Dobson, G. E. (1874). Notes on the respiration of some species of Indian freshwater fishes. Proc. zool. SOC.Lond. 1874
312-321.
Ghosh, E. (1934). An experimental study of the asphyxiation of some air-breathing fishes of Bengal. J. Asiat. Soc. Beng.
(N.S.) 29 327-332.
Hora, S. L. (1935). Physiology, bionomics and evolution of air-breathing fishes of India. Trans. nam. Inst. Sci. India 1:
1-16.
Hughes, G . M. & Munshi, J. S. D. (1973~).Fine structure of the respiratory organs of the Climbing perch, Anubus
testudineus (Pisces: Anabantidae). J . Zool., Lond. 170 201 -225.
Hughes, G. M. & Munshi, J. S. D. (19736). Nature of the air-breathing organs of the Indian fishes Channa, Amphipnous,
Clarius and Saccobranchus as shown by electron microscopy. J. Zool., Lond. 170 245-270.
Hughes, G. M. & Munshi, J . S. D. (1978). Scanning electron microscopy of the respiratory surfaces of Saccobranchus
(= Heteropneustes) fossilis (Bloch). Cell. Tiss. Res. 1 % 99- 109.
Munshi, J. S. D. (1960). The structure of the gills of certain freshwater teleosts. Ind. J. Zool. 1: 135-174.
Munshi, J. S. D. (1962~).On the accessory respiratory organs of Heteropnewles fossilis Bloch. Proc. R. Soc. Edin. ( 8 )
68: 128-146.
Munshi, J. S. D. (1962b). On the accessory respiratory organs of Ophicephaluspuncratw (Bloch) and 0.srriatus (Bloch)
J. Linn. Soc. Lond. (Zool.)44:616-626.
Munshi, J. S. D. (1976). Gross and fine structure of the respiratory organs of air-breathing fishes. In Respiration of
amphibious vertebrates: 73-104. Hughes, G . M. (Ed.). London & New York: Academic Press.
Munshi, J. S. D. (1983). The structure, function and evolution of the accessory respiratory organs of air-breathing fishes
of India. In Funcrional morphology oJ verrebrafes.Duncker, H. R. & Fleischer, G. (Eds). Stuttgart, N.Y.: Giessen,
1983, Gustav Fisher Verlag.
Munshi, J. S. D. & Singh, B. N. (1968). On the respiratory organs of Amphipnous cuchia (Ham. Buch.). J . Morph. 124:
423-444.
Satchell, G. H. (1976). The circulatory system of air-breathing fishes. In Respiration of amphibious vertebrates: 105- 123.
Hughes, G. M. (Ed.). London 19New York: Academic Press.
 ACCESSORY RESPIRATORY O R G A N S OF S N A K E - H E A D E D F I S H 317

Abbreviations used on figures and plates

AV Afferent vessel LO Labyrinthine organ
BC Blood capillary LAB Levator arcus branchialis
BM Basement membrane MC Mast cells
BV Blood vessel MV Microvilli
CB Ceratobranchial MR Microridge
cpc Cap cell N Nucleus
CT Connective tissue N V A Non-vascular area
CTC Connective tissue cell OP Operculum
EB Epibranchial Oes Oesophagus
EC Epithelial cell P Parasphenoid
EP Epithelium Ph Pharynx
EN Endothelium Rim Rim of receptacle of vascular papillae
ENC Endothelial cell RI Respiratory islet
EV Efferent vessel RBC Red blood cell
GA Gill arch SBC Suprabranchial chamber
GF Gill filament Sht Shutter
HB H ypobranchial VP Vascular papilla
HM Hyomandibula VL Valve
IA Inhalent aperture V Vein