Proc. Zool. Soc. lndia.

7 l2l 27 - 39 : (2008) =

OOCY'TE RESORPTION IN THE PYRGOMORPHID GRASSHOPPER POEKILOCERUS PICTUS FABRICIUS

SOMENATH DEY* AND M. RAZIUDDIN P.G. Department of Zoology, Vinoba Bhave University, Hazaribag 825 30'1, Jharkhand * Department of Zoology, Krishnagar Government College, Krishnagar, Nadia 741101, West Bengal Email: somenath0B@yahoo.co. in

Received - 10.04.2008
ABSTRACT

Accepted - 24.07.2008

This study was undertaken to investigate the phenomenon of oocyte resorption (oosorption) in the vitellogenic oocytes of Poekilocerus picfus. An appreciabty high percentage (45%) of the growing basal oocytes undergoes resorpflo n in both laboratory. reared and field-collected females. Light microscopy, scanning electron microsicopy and fransmissio n electron microscopy have been used to study the morphology and hi_stology of the resorption body.

Keywords

Oocyte, Resorptive bodles, Oosorption,

Grasshopper, SEM, TEM
INTRODUCTION

Resorption of oocyte has been reported in a number of insect orders (see Engelmann, 1970; Chapman,2001). This phenomenon is more pronounced in insects facing adverse conditions (Verma and Raziuddin, 1993, 1993a, 1g93b; Anwar and Raziuddin, 2002,2003). However, varying degree of resorption occurs in insects even under apparently optimal conditions. ln some acridids, a variable number of eggs which have grown to various sizes undergo degeneration while oocytes in the neighboring ovarioles continue to incorporate yolk and mature (Phipps, 1949, 1966; Singh, 195g; Highnam and Lusis, 1962; Lusis, 1963; Tobe and Pratt, 1975; Raziuddin et at., 1gg7; Karim, 1989). This phenomenon is also reported for many other insects (see Engelmann, 1970; Verma, 1991 ; Sharma, 1992).
A review of literature reveals that in the pyrgomorphid grasshoppe

r, Poekilocerus

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DEYAND RAZIUDDIN

pictus the phenomenon of oocyte resorption has been briefly described in the past by
'Raziuddin et at. (1987) and Anwar and Raziuddiil (2002, 2003). The present paper deals with a comparative description of oocyte resorption in field reared and laboratory reared Poekilocerus pictus as well as their morp,hshistology as revealed through light microscopy and electron microscopy.

MATERIALAND METHODS Live P pictus were collected from wild fields of both Jharkhand and West Bengal and reared in insect rearing cages at P.G. Department of Zoology, Vinoba Bhave University, Hazaribag. The insects were maintained on fresh Calotropis procera leaves. The rearing techniques described earlier by Raziuddin et al., (1977) and Raziuddin and Anwar (1996) have been followed. The female grasshoppers of known ages were dissected in insect Ringer solution at selected intervals and ovaries were examined after short fixation in formalcalcium (Highnam etal., 1963; Raziuddin ef a/., 1987) and numbers of resorptive bodies were noted. For the purpose of comparison of age between laboratory-reared and field-collected specimens, size of the vitellogenic basal oocytes was considered.
Resorptive bodies were fixed in aqueous Bouin's fluid for light microscopy. Paraffin (melting point 56-5BoC) sections of 6p thickness were cut by a rotary microtome (1090A, WESWOX, DPTIK). The sections were double stained by Delafield's haematoxylin and

eosin (alcoholic).
For scanning electron microscopy (SEM) resorptive bodies were fixed in 2.5 per

centglutaraldehydein0.l MphosphatebufferatpHT.4at4oCandthendehydratedby
ascending grades of alcohols (30%, 50o/o,70o/o,90% and absolute). These were then immersed in a mixture of alcohol and acetone of various grades (3:1 ,2:1, 1:1 , and 1:2) and finally dehydrated in anhydrous acetone at room temperature and proceeded.for critical point drying (CPD). After CPD samples were coated by gold by sputter (30 min) and then viewed under scanning electron microscope (JEOL JSM 6700F).

For transmission electrorr microscopy (TEM) resorptive bodies were primarily fixed in 2.5o/o glularaldehyde in 0.12 M Millonig's phosphate buffer for four hours at 4oC. Then they were secondarily fixed (post fixation) in 1% aquas osmium tetroxide for 15 minutes at 4oC and dehydrated by ascending grades of alcohols (30%, 50o/o,70o/o, 90o/o and absolute). Before embedding samples were immersed in transitional fluid (epoxypropane). Embeddings were done in araldite. Rough trimming were made by glass knives and then ultra thin sections (60nm) were obtained from the ultramicrotome (LKB Bromma with Olympqg,microscope). Ultra thin sections were collected on golden grids and the grids containing the sections were stained by uranyl acetate and lead citrate.
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PROC. ZOOL. SOC. tNDtA The grids were dried in desicator and then viewed under transmission electron'mlcroscope (FEl FP 5018/40 TECNAI G2 Spirit Bio Twin).
RESULTS

pictus paired ovaries lie dorsolaterally in the abdomen on either side of the alimentary canal. Generally the two ovaries of a specimen vary in length, the left ovary being larger than the right ovary and hence the former contained more number of ovario'les (105.66 t 9.6) than the right ovary (75.0 t 7.7) [Table l]. ln P. pictus 3 to 5 percent of the previtellogenic basal oocytes fail to incorporate yolk from the very beginning and remain as such throughout the ovarian cycle, while in the neighboring ovarioles yolk deposition had started and developed normally. After the process of vitellogenesis had started and oocytes had grown to various sub-mature sizes, some of the oocytes undergo oosorption or resorption and were finally resorbed. lt was, therefore, many resorptive bodies could be found in each ovary of the females in which oocyte maturation was nearing completion. Oosorption of growing vitellogenic oocytes has been observed in both field-collected as well as well fed and properly mated labbratory-reared females. The record of vitellogenic basaloocytes of P pictus undergoing oosorption is presented in Table ll. The observations

ln Poekitocerus

show that in both field-collected as well as laboratory-reared females' almost similar numberof oocytes underwentoosorption (average 81.16 in laboratory-reared and 82.8 in field-collected specimens) (Fig. 1). On the whole in P. pictus about 45 percent of basal oocytes do not mature and undergo resorption. Further, the percentage of vitellogenic

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DEYAND RAZIUDDIN basal oocytes which i.rnderwent resorption increased as the vitellogenesis proceeded. lt was found that the growing oocytes may

TABLE NO. I: NUMBER OF OVARIOLES IN LEFT AND RIGHT SIDE OVARY OF

LABORATO RY-REARED
PO

EKI LOCERUS

P I CT U S

Number of ovarioles

Left Ovary
SI
01

Right Ovary
SD S!
01

No. of ovarioles
126
109 95
115

Mean

No. of ovarioles
71

Mean

SD

02 03

02 03

62 59
B5

u
05 06 07 08 09
10
11

u
05

105

7B
61

%
103
101

{05.66

9.6032

06 07 08 09
10
11

75.0

7.6539

82
81

117 110

86 85 66

97 96

12

12

enter oosorption at any stage during their maturation but majority of the oocytes entered oosorpti6n when they were 3.0 to 5.5 mm long and only about 5 percent of the oocytes underwent oosorption when they were nearing maturation. lt was, therefore, the basal oocytes undergoing oosorption were in different stages of oosorption.

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PROC. ZOOL. SOC. INDIA

Table-ll: showing the number of resorptive body in taboratory-reared and field-collected female poekilocerus picfus.
sr. Laboratory-reared female
Left Ovary
01 V2

Field-collected femate

RightOvary
32 28 27 38 35

LeftOvary
59

RightOvary
35

57

49 43
52

47 45
50

30 26 40
37
31

03

M
05 06 07 08 09
10
11

47 42

49 45 45 47
52
51

27
37 36 39
38

6
45
53

30
36 37

49 44 42

40
31

30
3B

46 45

12

40

Range
Mean

42 -57
47.41

27

t SD

-39

45-59
48.41

26 -40

!4.53

33.75!4.47

3.89

34.41!4.49

Fig. 1: Comparison of oocytes undergoing

oosorption in left (LO) & right(RO)ovaries of

pictus

P.

BLab.

rcared
.--l

rE Field collected

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DEYAND RAZIUDDIN SURFACESTRUCTURE

Scanning electron microscopic studies of a few basal oocytes in different stages of oosorption revealed that the surface of such oocytes gets enormously folded (Fig. 2).
The folds are of various dimensions and are mainly longitudinal, some taking spiral course

(Fig

3).

Fig. 2: Scanning electron photomicrograph of resorptive body (RB) of P. pictus Showing folds over the surface. TR - trachea.

Fig. 3: Enlarged surface area of resorptive body showing spiral fold (SR).
The oocytes in advanced stage of oosorption clearly show that their surfaces are degenerated and damaged with the presence of a number of broken areas (Fig. 4). The surface bears a'number of blunt warts of various dimensions some of which have their surface cracked (Fig. 5). lt also appeared that the oosorbed materials are exuded-out in the form of granules of various shapes and dimensions (Fig. 6, 7).

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PROC. ZOOL. SOC. tNDtA

Fig. 4: SEM of resorptive body showing degenerated and damaged surface (.). Note the warts (W)over the surface.

Fig. 5: Highly magnified SEM photogiaph showing the cracks (c) over the resorptive
body surface.

Fig. 6: SEM showing exudations of various dimensions over the surface of resorptive body. Fig. 7: Magnified SEM showing blunt warts O over the surface of resorptive body which ultimately
break to release material.

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DEYAND RAZIUDDIN HISTOLOGY

Sections of the basal oocytes in advanced stage of oosorption when studied

under light microscope revealed the following features:

i) ii) iii) iv) v)

The follicular epithelium is disorganized but many of the cells still remain attached to tunica propria,
tunica propria is considerably folded, oocyte nucleus disorganized, decreased size of yolk spheres obviously because of their breakdown, and the cytoplasm is extensively vacuolated.
Studies by transmission electron microscopy also revealed extensive vacuolation

of cytoplasm and nuclearfragmentation (Fig.8,9 and 10).

Fig.8: Transmission electron micrograph of resorption body showlng many vacuoles (V) and nuclear fragments (.) in the cytoplasm. Fig. 9: TEM photomicrograph of resorption body showing vacuoles (V) in the cytoplasm (highly magnified).

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PROC. ZOOL. SOC. tNDtA

Fig. 10: Transmission electron micrograph of resorption body (highly-magnified) showing nuclear fragments (NF) scattered in the cytoplasm. DISCUSSION

Vitellogenic oocytes undergo resorption in many groups of insects. The oocytes enter resorption at any stage of their growth and thus resorption is not time and state bound event in the ovarioles. ln Poekilocerus plcfus the average number of ovarioles in the paired ovaries is 180 but a female has never been found to lay these numbers of eggs. Raziuddin ef al., 1977 has reported 66 to 136 eggs / pod in this insect. Lateron Anwar and Raziuddin (2003) have reported 114.60 t 3.81 eggs /pod in field-collected gravid females and 83.87 t 3.78 eggs / pod in laboratory-reared P. pictus females. Almost similar observations have been made by other workers viz., Pruthi and Nigam (1939), Menon (1952), Parihar (1974) in this insect. Variations in the number of eggs /pod described by different authors is'most likely due to difference in rearing conditions in different laboratories as well as difference in the environmental conditions prevailing in the fields in different regions. ln all the observations the number of eggs/ pod in P.pictus. is much less than the total number of ovarioles in the ovaries in a female. The difference in the total number of ovanoles in the ovaries of femalesand number of eggs laid by them is obviously due to the fact that many of the vitellogenic basal oocytes in the ovaries do not mature and are resorbed sometime during their development even though apparently optimal conditions prevailed in the laboratory or in the field. Under the most favourable conditions, in the desert locust, with about 50 ovarioles / ovary, nearly 22o/o of oocytes unciergo oosorption (Highnam and Hill, 1977). Similarly in females of Locusfa about 25% oocytes were resorbed even when these were reared on high quality food (see Chapman,
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DEYAND RAZIUDDIN
2OO1). tn autogenous females 'of Chrysomya (Screw-worm fly) 30% of the first batch of

oocytes was resorbed and even if these were reared on additional protein, 10% of the oocytes underwent oosorption (Spradberry and Schweizer, 1981).

The present results clearly demonstrate that abq$t- 45% of the basal oocytes underwent oosorption during maturation and thus only about 55% of the total ovarioles were able to mature and were subsequently laid by the females. Thus the percentage of oocyte resorption is appreciably high in P picfus in comparison to other insects described above. lt appears that the high percentage of oosorption i:n P. pictus which is almost similar in size to the desert locust is because of the presence of much higher number of ., ovarioles in the ovaries competing for nutrients and other factors during oocyte
development.

ln Locusfa the percentage of resorbed oocytes is inversely proportional to the quantity of the amount of grass eaten by the female (McCaffery, 1975). ln Cimex low levels of protein lead to resorption of oocytes (see Chapman,2001). ln the egg development of Aedes aegyptiresorption of oocytes depends on the quantity of blood and the interval between blood-meals (Lea et a|.,1978). Besides food, adverse conditions such as parasitisation of the insect (Liu, 1992), absence of proper mating (Willis ef a/.,
1958; Highnam ef a/., 1963; Anwar and Raziuddin; 2003), extremes of photoperiods (Verma and Raziuddin, 1993a, 1993b; Anwar and Raziuddin, 2002), age of the insect and inability to produce or lay fertile eggs (see Chapman, 2OO1), unfavourable conditions for egg

laying (Phipps, 1966) etc. also lead to resorption of oocytes. According to Highnam ef a/ (1963) in Schistocerca gregaria competition between the vitellogenic oocytes for available proteins and corpus allatum hormone play a key role in oocyte resorption, Resorption of a certain proportion of vitellogenic oocytes thus appears to provide a mechanism for increasing availability of nutrients and hormone (which facilitate vitellogenesis) to the remaining growing oocytes to ensure formation of eggs with adequate quantity qf yolk (Be!l and Bohm, 1975; Highnam and Hill, 1977). ln P. pictus marked morphological and histological changes occur in the oocytes undergoing oosorption. Light microscopical findings of resorption bodies of P. pictus revealthat the follicular epithelium becomes folded and disorganized and the cytoplasm becomes extensively vacuolated resulting in the shrinkage and collapse of the oocyte to form a resorption body. SEM and TEM studies confirm the above facts. ln the SEM study shrinkage, damage and di6oiganization of the surface are clearly revealed.'lt also appears that the materials fioh the resorbed oocyte are returned back to the haemolymph pool in the form of droplets which are to be utilized by other developing ogcytes.
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PROC. ZOOL. SOC. INDIA AGKNOWLEDGEMENTS The authors wish to express theiigratitude to the Director, lndian Association for the Cultivation of Science, Jadavpur, Kolkata and the Director, lndian lnstitute of Chemical Biology, Jadavpur, Kolkata for providing facilities of electron microscopy.
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and

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