In Crustacea, the distribution of NSCs is limited to the brain,

thoracic ganglia, circum-oesophageal connectives and eyestalk.

Enami (1951) first described different types of NSCs and mapped

them in the brachyuran crab, Sesarma dehaani

Panneerselvam, M

Subramoniam, T

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„>A»;S!£S " * • \ % 3 • U ^ ' CMFRI SPIXIAL PUBLICATION Number *> MANUAL OF RESEARCH METHODS FOR MARINE INVERTEBRATE REPRODUCTION Issued on the occasion of the Workshop on MARINE INVERTEBRATE REPRODUCTION joint ly organised by the Department of Zoology, Universi ty of Madras and the Centre of Advanced Studies in Mar icu l t i i re , Central Mar ine Fisheries Research Inst i tu te, Cochin held at the Universi ty of Madras f rom 25th October to IOth November 1982 The Centre of Advanced Studies in Mariculture was started in 1979 at the Central Marine Fisheries Research Institute, Cochin. This is one of the Sub-projects of the ICAR/UNDP project on 'Post-graduate agricultural education and research'. The main objective of the CAS in Mariculture is to catalyse research and education in mariculture which forms a definite means and prospective sector to augment fish production of the country. The main functions of the Centre are t o : —provide adequate facilities to carry out research of exce-llence in mariculture/coastal aquaculture ; —improve the quality of post-graduate education in mari. culture; —make available the modern facilities, equipments and the literature; —enhance the competence of professional staff; —develop linkages between the Centre and other Institutions in the country and overseas ; —undertake collaboration programmes ; and —organise seminars and workshops. Under the programmes of the Centre, post-graduate courses leading to M.Sc. (Mariculture) and Ph.D. are offered in colla-boration with the University of Cochin since 1980. Front cover: SEM picture showing surface topography of Strepto-cephalus dichotomus egg. Manual of Research Methods for Marine Invertebrate Reproduction EDITED BY T. SUBRAMONMM Unit of Invertebrate Reproduction, Department of Zoology, University of Madras, Madras-600 005 & CMFRI SPECIAL PUBLICATION Number 9 ISSUED ON THE OCCASION OF THE WORKSHOP ON MARINE INVERTEBRATE REPRODUCTION JOINTLY ORGANISED BY THE DEPARTMENT OF ZOOLOGY, UNIVERSITY OF MADRAS AND THE CENTRE OF ADVANCED STUDIES IN MARICULTURE, CENTRAL MARINE FISHERIES RESEARCH INSTITUTE HELD AT THE UNIVERSITY OF MADRAS FROM 25TH OCTOBER TO 10TH NOVEMBER, 1982 . (LIMITED CIRCULATION) Published by: E. G. SILAS Director Central Marine Fisheries Research Institute Post Box No. 1912 Cochin 682 018 PRINTED IN INDIA AT THE DIOCESAN PRESS, MADRAS'7—1982 C3626 PREFACE The technologies of controlled reproduction, induction of spawning, sex reversal, artificial fertilisation, sterilisation and preservation of gametes are increasingly applied in aquaculture to obtain quality seed, quality fish stock and better yield. In this context, researches on different aspects of reproduction, developmental biology and physiology have assumed consider-able importance besides their values in understanding of the ontogeny of the organisms. Extensive researches carried out in recent years from several laboratories in the world have not only accumulated a body of information, but also broughtforth several new concepts to our understanding of the development and reproductive behaviour of finfishes and shellfishes-In India, directed research on reproductive physiology and biology is taken up only recently and the field is still in an infant stage. In view of its emerging importance, it is identified as an area for priority research and for expertise development in the programmes of the Centre of Advanced Studies in Mariculture at the Central Marine Fisheries Research Institute, and several programmes of research are being taken up in this field with particular reference to the reproductive behaviour of the culti-vable finfishes and shellfishes. Advances made on the frontiers of invertebrate reproduction in recent years have been significant enough to organise a national workshop and to prepare a manual on research methodologies for the study of the subject. Several histological, histochemical and biochemical methods and sophisticated instru-ments have been introduced in these studies making it essential that the scholars who desire to work and specialise in the field are given adequate basic information on the research methods so as to enable them to appreciate and advance research to understand the problems confronted in the field. The present manual, the third in the series, is prepared and compiled by Dr. T. Subramoniam, Leader of the ' Unit of i i i Invertebrate Reproduction ' of the Zoology Department of the University of Madras, Tamil Nadu. During the past decade, a team of research scholars are working on different aspects of marine invertebrate reproduction including the cultivable crustaceans such as Scylla serrata, Panulirus homarus and Macrobrachium spp. under his leadership. Contributing to our knowledge on the subject, the research results achieved so far in these aspects by the Unit have unfolded several new concepts in oogenesis, spermatogenesis, sperm transfer strategy, ferti-lization and endocrine control of reproduction and gamete formation. I wish to express my great appreciation to Dr. T. Subramoniam and his team of Scholars, who by their dedication and interest evolved a series of tested research methods and set a theme of investigation through insight and skill on marine invertebrate reproduction. I am sure that this manual will be of immense use to the research scholars and scientists who would like to specialise in the subject and cognate fields. This is the second workshop we are organising in close colla-boration with the University of Madras. I wish to express my gratitude to Dr. M. Santappa, Vice-Chancellor, University of Madras for the keen interest evinced in such collaborative pro-grammes and for the advice. I am also indebted to Dr. K. Ramalingam, Professor and Head of the Department of Zoology, University of Madras for productive discussions, continuous support and suggestions. I wish to thank Shri P. T. Meenakshisundaram and Shri K. Rengarajan, Scientists of the Central Marine Fisheries Research Institute for their help in the preparation of this manual. E. G. SILAS, Director, C.M.F.R.I. iv 15 A CLASSIFICATION OF NEUROSECRETORY CELLS OF CRUSTACEA* 15.1. INTRODUCTION In Crustacea, the distribution of NSCs is limited to the brain, thoracic ganglia, circum-oesophageal connectives and eyestalk. Enami (1951) first described different types of NSCs and mapped them in the brachyuran crab, Sesarma dehaani. Following this, there were numerous reports on the classification of NSCs in a variety of crustaceans belonging especially to the superior orders (Durand, 1956 ; Matsumoto, 1958 ; Adiyodi and Adiyodi, 1970). One of the compelling inference from these studies is that the morphology of the NSCs in different forms is highly variable and species-specific. In the present experiment a classification of NSCs of an anomuran crab is attempted employing various criteria such as size, shape, location, tinctorial properties and cytological details. 15.2. MATERIAL Brain and thoracic ganglion of the anomuran crab, Albunea symnista. 15.3. PROCEDURE 1. Observe the slides prepared in the Expt. No. 14. 2. Note the size, shape, location and other cytological details of the NSCs. 3. Classify the NSCs into different types based on the char-acteristics mentioned in the step 2. 4. Draw the camera lucida drawings of NSCs (Fig. 1). * Prepared and verified by M. Panneerselvam and T. Subramoniam, Unit of Invertebrate -Reproduction; ©epattnwtrt^  ef-'Zeelogy, • University of Madras, Madras-600 00$. i l l Type A Type A' Type B ..../tVy-A-> < ^ ' J W . ' Type C Type D Type A Type 6 Type C Fig 1. Types of N§C8in,tlnbt»in^o)aad thoracic gsmglion (6) oiA. symustd} 43$ 5. Map the distribution of NSCs in different regions of the brain and thoracic ganglion. For this, first draw the outline of intact neuroendocrine centres. By using various symbols, indicate the location of individual NSCs as shown in Figs. 2 and 3. 15.4. OBSERVATIONS 15.4.1. NSC types of the brain (Fig. la) A type NSC: These NSCs are 45 fj.m-50 pm in diameter. The nucleus is oval in outline and possesses two nucleoli. The cytoplasm is flaky due to the accumulation of secretory granules. These cells are distributed in the dorsal and ventral median regions of the protocerebrum. The axons are indistinguishable. A' type NSC: It is similar to the A type NSC, except for the presence of an axon. Cells of this type are mainly located along with the A type in the dorsolateral and ventromedian regions of the protocerebrum. B type NSC: These cells are oval and 35 fWrt-40 pm in diameter. The nucleus is prominent, but without a distinct nucleolus. These cells are known to be distributed in ; the anterolateral region of the protocerebrum with a few ceils scattered in the ventrolateral region. As in the A type, axons are wanting. C type NSC: These are conical or club shaped, and are few in number, measuring 25 urn - 30 nm in diameter. They are distributed in the dorsal and ventral regions of the deutocere-brum. A few cells are also seen near the A type. D type NSC: This type is spherical without an axon, and mea-sures 8 nm -10 nm in diameter. The cytoplam is scanty, but possesses dark blue granule. Cells of this type are confined exclusively to the dorsal region of the protocerebrum. 15.4.2. NSC type of the thoracic ganglion (Fig. lb) A type NSC: It is monopolar, uninucleolated and 25 pm-30 jim in diameter. The shape of the nucleus varies from flat to crescent. These cells are located in the dorsal and ventral regions of the neuropile. 137 10 LL ELL ELL E L L CE LL OF THORACIC GANGLION Fig. 2. Distribution of NSCs in the brain of A. symnista. (a) dorsal side, (b) ventral side. I—optic nerve, II—Antennulary nerve, III—Antennary nerve, IV—Maxillary nerve, V—Circum-oesophageal connective. Fig. 3. Distribution of NSCs in the thoracic ganglion of A. symnista. (a) dorsal side, (b) ventral side. I and IV—Thoracic nerves, III and IV—Nerves to thoracic appendages. 138 B type NSC: These cells are oval shaped, more abundant and measure 15 nm-20 pm in diameter. Cells of this type are binucleolated and found distributed in the dorsal and ventral regions of the thoracic ganglion. A few cells are also noticeable among the A type NSC. C type NSC: This type NSC is round with a diameter of 10 fim-15 nm and distributed in the dorsal side of the anterior region. Tabulate the observations in the table given below: NSC types size shape location tinctorial cytological 15.5. REFERENCES ADIYODI, K. G. AND R. G, ADIYODI, 1970. Endocrine control of reproduction in decapod Crustacea. Biol. Rev., 45: 121-165. DURAND, J. B. 1956. Neurosecretory cell types and their secretory activity in the crayfish. Biol. Bull. (Woods Hole), 111: 62-76. ENAMI, M. 1951. The source and activities of two chromatophoro-trophic hormones in the crab of the genus Sesarma. II—Histo-logy of incrementary elements. Ibid., 101: 241-258. MATSUMOTO, K. 1958. Morphological studies on the neurosecre-tion in crabs. Biol. J. Okayama Univ., 4 : 103-186. 139 

A classification of neurosecretory

cells of crustacea

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A classification of neurosecretory cells of crustacea

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