The Development of the Spermatophoric Mass of the RockLobster~ Parribacus antarcticus (Lund) 1

DONALD C. MATTHEWS2

INTRODUCTION

THE LITERATURE on the spermatophores ofthe tribe Scyllaridea is both superficial andscattered. Although cursory accounts of theextruded mass have appeared for both thePalinuridae and the Scyllaridae, few thoroughhistological studies have been made. Thispaper proposes to extend the knowledge ofthe biology of the common Hawaiian scyl­larid, Parribacus antarcticus (Lund), through anaccount of, its spermatophoric development.

Although CaIman (1909: 290) in discussingthe spermatophores of Crustacea fails to men­tion specifically the spermatophores of thePalinuridae [sens. lat.]' he does state that inmost Macrura the spermatozoa are enclosedin a sheath, forming a continuous cord-likemass. Moreover, he (loc. cit.) states that, "InScy//arus and in the Anomura this is brokenup into separate spermatophores attached byone end in a row on a strip of membrane."

Andrews (1912: 178), in referring to thespermatophoric mass of Panulirus argus La­treille, states:

When cut the interior was' white and full of holes ortubes. It suggested a very stiff cheese, but the minutecavities were not closed vesicles bur shorr tubes ... acareful examinarion of sections shows rhe short, tubularand somewhat branched cavities to be about one halfmillimeter in diameter and 1 or two long, separated byvery fine walls of the dense white rubber-like material... the enveloping mass is white except on the surfacewhere it is more or less black or grey according, ap­parently, to rhe length of exposure to water.

1 Contribution No. 41, Hawaii Marine laboratory.2 Deparrment of Zoology and Entomology, Uni­

versityof Hawaii. Man'uscript received December 22,1952.

28

Although the change in consistency and colorconforms to that observed by the author inthe extruded mass of Panulirus penici/latus(Oliver), the observed "tubular cavities" wereprobably merely the result of sections cutthrough a highly contorted, condnuous tube(Matthews, 1951: 365, fig. 10).

Andrews' description (oP cit., 183) of theexternal spermatophoric mass of Parribacusantarcticus is adequate, but again his refer­ence to an "internal . . .matrix enclosingtubular cavities" is, as will be shown subse­quently, erroneous.

Allen (1916), Fasten (1917), and Wilson(1948) described the change of color and con­sistency in the extruded spermatophoric massof Panulirus interruptus, but did not investi­gate its development within the vas defere~s.

The observations of Crawford and De Smidt(1923) on Panulirus argus were also limitedto the extruded spermatophoric mass.

The nonpedunculate spermatophore of Pa­nulirus penicillatus (Oliver) (vide Matthews,1951) resembles the spermatophores of theMacrura, but apparently no published ac­counts have verified CaIman's statement (op.cit.) regarding the pedunculate spermato~

phores of the Scyllaridae.

METHODS AND TECHNIQUES

Twenty sexually mature Parribacus antarc­ticus ,,(Lund) obtained from the HonoluluAquarium during May, June, and July, 1952,were used in this study. Of the 18 male re­productive systems dissected, 6 were used tostudy the nature of the testes and the coiledvasa deferentia, 8 were vitally stained, and 4

Spermatophoric Mass of the Rock Lobster - MArrHEws 29

were serially sectioned. Of the two femalesobtained, only one possessed a spermato­phoric mass; this was dissected and studied.

All dissections were made on living, non­anesthetized specimens immersed in sea wa­ter. The carapace was opened quickly, and theinternal organs overlying -the reproductivesystem were removed. The tes,tis and coils ofthe vas deferens were then studied andsketched.

-Two methods were employed in vital stain­ing with aqueous solutions of neutral red andtoluidin blue: (1) The stains were injectedinto the enlarged distal portion of the vasdeferens (Fig. 1d) and allowed to permeate

FIG. 1. Dissection of a mature male Parribacus ant­arcticus (Lund), drawn to show the position of rhereproductive system. a, Transverse bridge; b, testis;c, coiled, proximal portion of vas deferens; d, straight,distal portion of vas deferens; e, hyaline line. (O.6X.)

its contents proximally to the testis; (2) thevas deferens was removed from the body andplaced in the stains. Both neutral red andtoluidin blue were used in concentrations of1:1,000 to 1:10,000. For the injection tech­nique, solutions of approximately 1:1,000 to1:5,000 were used; for the immersion tech­nique, solutions of approximately 1 :10,000were used, and the vas deferens was allowedto remain in the stain from 10 to 30 minutes.The vas deferens was then removed from thestain and placed in sea water for subsequentstudy. Because injection tends to distend thewalls and causes the stains to permeate the

.surrounding tissues, this technique was laterabandoned in favor of the more easily con-trolled immersion method.

The walls of the vasa deferentia were thenteased open under a dissecting microscopeand the vitally stained contents studied anddrawn.

The vasa deferentia to be sectioned wereremoved quickly and placed in H(min's fluid.After fixation, the tissues were washed inalcohol (70 per cent) and either cleared indioxan and embedded in TisSuemat, clearedin toluene and embedded in Tissuemat, orplaced in dioxan, cleared in methyl-benzoate(1 per cent celloidin added), washed in xylene,and embedded inTissuemat (54-56°C.). None _of these modifications was completely satis­factory for all regions of the vas deferens.All gave good preparations for sectioning thesmall proximal regions and poor preparationsfor sectioning the large, distal regions. Thiswas due to the presence of the matrix (Fig.10c) which upon dehydration had becomeextremely brittle. The distal region of the- vasdeferens could be sectioned only after im~mersing the block in water, although thisresulted in some swelling of the connectivetissue. The tissues were sectioned at 10 mi­crons, stained in standard alumhaematoxylin,and counterstained with eosin.

The writer wishes to thank Mr. SpencerTinker for supplying the specimens and Mr.James Park for making the illustrations.

30 PACIFIC SCIENCE, Vol. VIII, January, 1954

FIG. 2. A small portion of the testis and proximalvas deferens showing results of toluidin blue immer­sion technique. a, Sacculi; b, undifferentiated sperma­togenic substance; c, moniliform portion; d, capsuleof ampulla; e, distinct ampullae; f, thin wall of vasdeferens. (15 X.)

sacculi open (Fig. 2a). The sacculi of differentregions appear indistinguishable as to numberand size. Both testes are enveloped by adelicate mesentery and penetrated by numer­ous blood vessels.

The vas deferens (Fig. Ie) which emanatesfrom the testis is an extremely small, highlycoiled.tube which, increasing in diameter dis­tally, finally emerges as a large tube (d) open­ing on the coxopodite of the fifth periopod.A hyaline line (e) traverses most of the lengthofthe otherwise chalky, opaque tube, markingthe position of the internal typhlosole (Fig.lOd). This hyaline line is prominent only inthe distal enlarged portion (Fig. Id). The

DISCUSSION

FIG. 3. A small portion of the vas deferens imme­diately distal to the region illustrated in Figure 2,showing results of toluidin blue immersion technique.a, Secretion which envelops the already encapsulatedampullae; b, sheath between successive ampullae. (15 X.)

FIG. 4. Diagram of a small portion of the vas deferensimmediately distal to the region illustrated in Figure 3,showing the results of immersion technique. a, Suc­cessive encapsulated ampullae; b, sheath surrqundingencapsulated ampulla; b' , ribbon-like sheath betweensuccessive ampullae; c, tWisted, ribbon-like sheath.(15X.) -

numerous coils of the proximal portion arenot constant either as to number or direction _of their turns, as none of the vasa deferentiastudied was identical.

All regions of the vas deferens fail to exhibitmuscular contractions and are quiescent evenwhen exposed quickly. There were no inter­mittent contractions which are so character­istic of certain vasa deferentia. Whereas thepersistent mobility of the vas deferens ofDardanus asper (Matthews, 1953: 258) may berelated to its stamina, the apparent immobil­ity of the vas deferens of P. antaretieus may berelated to its debility. The cessation of con­tractions in the altered, exposed vas deferensobscures the mechanical aspect of spermato­phoric development. Vital staining is there-

ba

The testes (Fig. Ib), when freed from over­lying tissues, resemble those of Panuliruspeni­cillatus (Matthews, 1951: 360) in that theyform an elongate H, the right and left sidesjoined by a transverse bridge (a) just posteriorto the junction of the pyloric region of thestomach to the intestine. As in P. penieillatus,the testes lie contiguous to the digestive tract.Occasionally one, less frequently both, of theportions of the testes anterior to the bridgebifurcate, although the posterior region isunaltered. The dissecting microscope revealsthe testis as a highly coiled, continuous, thin­walled tube into which innumerable small

Spermatophoric Mass of the Rock Lobster - MATTHEWS 31

FIG. 5. Diagram of 'a dissecred portion of the vasdeferens immediately distal to the region illustrated inFigure 4, showing the results of immersion technique.a, Widely separated encapsulated ampullae; .b, highlytwisted, ribbon-like sheath. (15 X.)

fore invaluable in discerning the effects ofcontractions in the unaltered vas deferens.

Figure 2 illustrates the results of immersinga small pottion of the testis and the proximalregion of the vas defetens in toluidin blue.Here the thin wall of the vas deferens (f)affords rapid penetration of the stain, and theabsence of the matrix (Fig. lOe) makes clearthe developing spermatophore. The undiffer­entiated spermatogenic substance (Fig. 2b)is seen as it enters the vas deferens. As ittraverses the vas deferens, this undifferentiated,spermatogenic substance becomes moniliform(e) and, acquiring a capsule (d), separatesinto distinct ampullae (e).

dcba

v

FIG. 6. Diagram of a dissected portion of the vasdeferens distal to the region illustrated in Figure 5.a, "Elevated" ampullae; b, entwined, twisted ribbons.(15X.)

FIG. 7. A cross section through the vas deferens inthe region indicated by b of Figure 2. a, Undifferen­tiated spermatogenic mass; b,' circular lumen; c, wall,of vas deferens; d, epithelial layer. (134X.)

and the connecting sheath (b) is more highlytwisted.

A diagram (Fig. 6) of a dissected portionof the vas deferens distal to Figure :5 showsthe ampullae (a) less widely separated and"elevated" by the entwining of their alreadytwisted connecting sheaths (b).

Serial sections substantiate what vital stain­ing has so far revealed and enable one todetermine the origin of the various secretions.Figure 7 is a cross section through the vasdeferens in the region indicated by b of Figure2. The undifferentiated spermatogenic mass(a) occupies the almost circular lumen (b).The wall of the vas deferens is here composed

ha

Figure 3 illustrates the results of the im­mersion technique on a small portion of thevas deferens immediately distal to the regionillustrated in Figure 2. A new sheath (a) nowenvelops the already encapsulated ampullaeand occupies the space between successiveampullae (b).

The results of the immersion technique ona small portion of the vas deferens immedi­ately distal to the region illustrated in Figure3 are shown in Figure 4. The successive am­pullae (a) are now more widely separated.The sheath (b) which envelopes the alreadyencapsulated ampullae is thin and betweensuccessive ampullae is now ribbon-like (b').In the distal portion of this region the ribbon­like sheath connecting successive ampullaeis twisted (e).

Figure :5 is a diagram of a dissecte<1 portionof the vas deferens immediately distal to Fig­ure 4. The ampullae (a) are widely separated

32

FIG. 8. A cross section through the vas defetens in-the region indicated by c of Figute 2. a, Crypt-likefold of the epithelium; b, spermatids; c, secondaryspermatocytes; d, capsule secretion; e, primary sper­matocytes. (106X.)

of a thin muscular and connective tissue layer(c), and a thin layer of epithelium (d). InFigure 8 the spermatophoric capsule is beinglaid down by a secretion (d) of the crypt-likefolds of the epithelium (a). It is significantthat spermatogenesis is taking place. Primaryspermatocytes (e), secondary spermatocytes(c), and spermatids (b) are clearly discernible.

Figure 9, a cross section through the vasdeferens in the region indicated by a of Figure3, shows the secretion of the sheath (b) aroundthe already encapsulated ampulla: (c).

Figure 10 is a stereogram of the vas deferensthrough the region indicated by d of Figure1. The ampullae (a) are "elevated" and thealready twisted connecting ribbon-like sheathsare entwined (b). The matrix (c), the copioussecretion of the typhlosole (d), has forced thespermatophores to lie opposite the hyalineline (e).

Figure 11 is a drawing of a portion of afemale sternum (a) and a partially dissectedspermatophoric mass. The hard, darkenedouter portion of the matrix (b) envelops theless hardened matrix (c) which contains theampullae (d) attached to doubly - twistedpeduncles.

PACIFIC SCIENCE, Vol. VIII, January, 1954

The undifferentiated sperm mass emanatesfrom the testis probably because of the con­tinuous activity of the germinal epitheliumlining the sacculi. In the proximal portion ofthe vas deferens, not more than 4 or 5 milli­meters long, this undifferentiated spermato­genic material is both moved along andmolded into a continuous moniliform mass,presumably by the muscular contractions ofthe wall. In this same region of the vas defer­ens the crypt-like folds of the epitheliumproduce a secretion which forms the thickcapsules of the now distinct ampullae.

As the encapsulated ampullae traverse thevas deferens, the epithelium produces anothersecretion, the sheath, which both surroundsthe already encapsulated ampullae and fillsthe intervening spaces. Less than 1 centimeterof the proximal vas deferens is traversed be- ­fore the portions of this sheath lying betweenadjacent ampullae are compressed into a thin,delicate ribbon, probably by contractions ofthe wall of the vas deferens. Because of theimmobility of the dissected vas deferens, theprocess was not observed.

The chalazal twisting of the connectingribbons probably is due to the rotations of the

a

FIG. 9. A cross section through the vas defetens inthe region indicated by a of Figure 3. a, Epithelium;b, sheath secretion which surrounds the encapsulatedampullae; c, capsule. (125 X.)

Spermatophoric Mass of the Rock Lobster - MATTHEWS 33

FIG. 11. A partially dissected spermacophoric masstaken from the sternum of rhe female. a, Portion ofsternum; b, hard, dark portion of matrix; c, soft, lightportion of matrix; d, ampullae attached co doublytwisted peduncles. (3.5 X.)

It should be recalled that only one femalewas observed which possessed the spermato­phoric mass. The actual process of spermliberation was not observed, but the scratched,hardened matrix, together with the special­ized claws (vide Andrews, op. cit.), suggest amechanical method.

The results of this investigation may leadto speculations on the systematic position ofthe Scyllaridae. Although the spermatophoresof Parribacus antarcticus are intermediate be­tween those of the nonpedunculate spermato­phores of a palinu~id (Panulirus penicillatus)and the pedunculate spermatophores of apagurid (Dardanus asper) , an inference thatParribaczts antarcticus is intermediate between

. these forms is not warranted. Anatomical, em­bryological, and serological evidence shouldaccompany such spermatophoric evidence,and this should be drawn from many speciesrepresenting many genera.

The present study on the spermatophoresot P. antarcticus should serve to question theinterpretation placed on the function of thepeduncle. The origin of the seemingly sig­nificant peduncle may have been only thefortuitous occurrence of mechanical and phy­siological activities of the vas deferens withlittle regard for the elevation' of the ampullaeand the subsequent dispersal of the sperma­tozoa. Certainly, in P. antarcticus the peduncleserves no such useful purpose, inasmuch asthe hard enveloping matrix prevents the am­pullae from being held "aloft."

ampullae as the coils of the proximal region .of the vas deferens are traversed. The "draw­ing together" of the ampullae probably is alsothe result of the shortening of the ribbons dueto this twisting. Moreover, the entwining ofthe already twisted ribbons to form the stalkor peduncle is also a result of this process.

The secretion of the typhlosole now sur­rounds the already formed spermatophoresand by its accumulation forces the continu­ous, pedunculate spermatophores against thewall of the vas deferens opposite the hyalineline.

As the spermatophores traverse and finallyaccumulate in the enlarged distal portion ofthe vas deferens, the process ofspermato­genesis is completed. The primary spermato­cytes have undergone two successive divi­sions, and the spermatids have metamor­phosed into mature spermatozoa.

It is at this stage of development that thespermatophoric mass is placed on the sternumof the female and, as described by Andrews .(op. cit.), darkens and hardens when it is ex­posed to sea water. The dissected spermato­phoric mass reveals not a series of tubularcavities but a continuous thread-like ribbon(or two ribbons if both vasa deferentia haveparticipated) from which the pedunculatespermatophores extend.

FIG. 10. Stereogram of the VlJ.S deferens in the regionindicated by d of Figure 1. a, "Elevated" ampullae;b, entwined, twisted ribbons; c, matrix; d, typhlosole;e, hyaline line. (25 X.)

34

REFERENCES

ALLEN, B. M. 1916. Notes on the spiny lob­ster (Panulirus interruptus) of the Californiacoast. Calif Univ., Pubs., Zool. 16(12):139-152.

ANDREWS, E. A. 1912. Spermatophores andspecialized claws in Palinurus and Parri­bacus. Zool. Jahrb., Supp!. 15 [vol. 3]:177-190.

CALMAN, W. T. 1909. Crustacea. In: A trea­tise on zoology. [Sir Ray Lankester Ed.] PartVII, fase. 3, 332 pp. A. & C. Black, London.

CRAWFORD, D. R., and W. J. J. DE SMIDT~

1923. The spiny lobster, Panulirus argus ofsouthern Florida-its natural history and

PACIFIC SCIENCE, Vol. VIII, January, 1954

utilization. U. S. Bur. Fisheries, Bu!. 38:281-310.

FASTEN, N. 1917. Male reproductive organsofDecapoda, with special reference to PugetSound forms. Puget Sound Mar. Sta., Pub.1: 285-307.

MATTHEWS, D. C. 1951. The origin, develop­ment, and nature of the spermatophoricmass of the spiny lobster, Panulirus peni­cillatus (Oliver). Pacific Sci. 5(4): 359-371.

---1953. The development of the pedun­culate spermatophore of the hermit crab,Dardanus asper (De Haan). Pacific Sci. 7(3):255-266.

WILSON, R. C. 1948. A review of the SouthernCalifornia spiny lobster fishery. Calif Fishand Game 34(2): 71-80.,