Oogenesis in the tropical land crab,Gecarcinus lateralis (Freminville)

Summary The ovaries of small and large adult Gecarcinus were studied histologically and histochemically at various stages in the annual cycle. At all seasons of the year, dividing cells are seen within germinal nests in the ovary. Following division, the cells within the germinal nest enlarge and appear to move out into the stroma, forming cords of young oocytes that become encapsulated by follicle cells. Glycogen, not demonstrable in cells within the germinal nests, is present in the perinuclear cytoplasm of both young and mature oocytes. Lipid is distributed peripherally in the cytoplasm of the oocytes. Deoxyribonucleoprotein is demonstrable within the nuclei of germinal nest cells and of the young oocytes; it is not detectable within the nuclei of the large oocytes. The histological observations suggest that oogenesis occurs throughout the reproductive life of Gecarcinus.Dedicated to Professor Berta Scharrer on her 60th birthday in love, respect and admiration. — This work was supported in part by U.S.P.H.S. Training Grant GM-102.I express my thanks to the late Dr. Helen W. Deane and Dr. Dorothy E. Bliss for their help and encouragement.     

Opioid potentiated chromatophorotropin regulation of pigment migration in the land crab Gecarcinus lateralis

In Gecarcinus lateralis dopamine treatment results in dispersion of black and concentration of red pigments within chromatophores. These effects of dopamine on the migration of pigments can be blocked by the dopamine antagonist haloperidol. These results strongly indicate the presence of a dopamine receptor mediated system in this organism. Serotonin injections also result in the dispersion of black pigment; however, this effect cannot be blocked by haloperidol. Norepinephrine was found to be without effect on this pigment regulatory system. Injections of crude eyestalk extract results in pigment migration within the chromatophores in both stalked and destalked animals. Injection of the stable methionine enkephalin analog FK 33 824 into the organisms causes no observable effects on the pigment system. However, coinjection with eyestalk extract strongly potentiates the effect of the extract. This potentiation can be completely blocked by the opiate antagonist naloxone, thus indicating that an endogenous opioid system may be part of the overall regulation of pigmentation movement.     

Fine Structure of Salt and Water Uptake in the Land-Grab, Gecarcinus lateralis

The salt-absorbing tissue is found in the respiratory lamellaeof the gills in the form of a highly interdigitated epithelium.The folds of the epithelium are supplied with mitochondria inthe form of "mitochondrial pumps." Intercellular spaces betweenthe folds also satisfy the morphological requirements for Diamond'smodel system for water transport based on his theory of standingosmotic gradients. Most of the osmoregulatory tissue is localizedin the three most posterior gills, which in turn rest on thepericardial sac. It is suggested that the pericardial sac transfersground water to the gills for salt and water absorption. Thepericardial sac serves for storage of water and as a hydraulicassist during the molt.     

The Foregut of Gecarcinus lateralis as an Organ of Salt and Water Balance

The permeability of the foregut of the land crab, Gecarcinuslateralis, to tritiated water (THO), Na²², and Cl³⁶ was studiedin vitro during the intermolt period and after ecdysis. In crabswith eyestalks, the foregut is permeable to water and ions inthe direction hemolymph-to-lumen and lumen-to-hemolymph, bothduring the intermolt period and after ecdysis. However, theforegut of animals without eyestalks is impermeable after ecdysis. The movement of THO always follows the movement of Na²² acrossthe wall of the foregut, while the movement of Cl³⁶ may or maynot be correlated with the movement of Na²² and THO. Comparisonof the ratio of water to ions in the hemolymph with the ratiocalculated from radioisotope flux indicates that Na⁺ and waterare probably moving isosmotically, although not necessarilyaccompanied by Cl^– When an extract of the thoracic ganglionic mass of G. lateralisis added to the "hemolymph side" of the foregut in vitro, thereis immediately a large increase in permeability to water andsalts. This occurs in the foregut of crabs with eyestalks duringintermolt and also in eyestalkless crabs after ecdysis. Thus, not only is the foregut of Gecarcinus lateralis permeableto water and salts in both directions, but also the extent ofits permeability is under neuroendocrine control. As a consequence,the animal may be able to move water and salts into the foregutor out of it into the hemolymph as needed. This may be an importantadaptation for a terrestrial crab that must conserve water,especially at the critical time of ecdysis.     

Transport of Oxygen by the Blood of the Land Crab, Gecarcinus lateralis

Parameters relating to transport of oxygen were measured inthe pericardial blood and venous outflow from the first walkingleg of Gecarcinus lateralis. O₂-equilibrium curves of the hemocyaninof G. lateralis were found to be sigmoid and, at 27°C andpH 7.45, to have a half-saturation pressure of about 17 mm Hgoxygen. Average partial pressures of oxygen as measured by O₂-electrodewere 32 mm Hg in pericardial blood and 9 mm Hg in the venoussamples. Analysis of the O₂-content in corresponding samplesby the Van Slyke technique revealed an average of 2.17 volumes% O₂-capacity for whole blood, 1.45 volumes % for pericardialblood, and 0.61 volumes % for venous blood. Estimates basedon the Van Slyke analyses indicated an average pO₂ of 29 and14 mm Hg in pericardial and venous samples, respectively. Thesefigures agree fairly well with those obtained by means of O₂-electrodes.Of the oxygen carried to the tissues, about 94% is carried asoxyhemocyanin and about 6% is carried in physical solution.As the blood passes through the gills, the hemocyanin, on anaverage, becomes 80–85% saturated with oxygen and returnedfrom the tissues 18–45% saturated with oxygen. These resultsindicate that the blood of G. lateralis has a higher O₂-capacitythan the blood of most other decapod crustaceans for which similarinformation is available. In addition, the blood of G. lateralistransports more oxygen to the tissues per unit volume than doother crustacean bloods.     

Water Balance in the Land Crab, Gecarcinus lateralis, During the Intermolt Cycle

Gecarcinus lateralis can take moisture from a clamp substratumin amounts adequate for the needs of the entire intermolt cycle.It can also rehydrate in this way, even after severe dehydration. This crab is able to survive for many months when free waterof a wide range of salinities (0.30; = parts per thousand)is made available in a shallow dish. The crab dies within sevenweeks when the salinity of this water is 35. During proecdysisthe pericardial sacs of eyestalkless crabs become most swollenwhen the salinity of the available water is 15 or 23, and survivalduring and after ecdysis is greatest with water of 15. A crab in proecdysis shows no increase in the rate at whichwater enters following dehydration. Yet large amounts of waterare retained, particularly at the intermediate salinities. Maximalswelling of the pericardial sacs just prior to ecdysis is essentiallyequivalent in crabs with eyestalks, in eyestalkless crabs, andin eyestalkless crabs that have received an implant of centralnervous tissue. Hence, we conclude that a hormone causing theretention of water exists, but not in the eyestalks, in thebrain, or in the thoracic ganglionic mass. At ecdysis eyestalkless crabs show large increases in the dimensionsof the carapace, while crabs with eyestalks and eyestalklesscrabs that have received an implant of certain central nervoustissues show much less increase and may even show a decrease.Thus, we conclude that a hormone causing a release of waterat ecdysis is produced in the central nervous system. The advantages to the crab of a dual hormonal control of itswater balance are discussed.     

Interruption of proecdysis by autotomy of partially regenerated limbs in the land crab, Gecarcinus lateralis

In the land crab, Gecarcinus lateralis, autotomy of partially regenerated limbs before a critical stage in the premolt period results in (1) a very rapid decrease in the serum ecdysone titer, (2) a delay in the growth of partial regenerates remaining on the animal, (3) a delay in the deposition of gastroliths, and (4) a delay in cytological changes in the epidermis. Serum ecdysone titers remain low while new limb regenerates form at the sites of those removed. Ecdysone titers rise when these secondary regenerates complete basal regeneration. Premolt events, which had ceased at the time of autotomy of the partial regenerates, resume their normal patterns of development when ecdysone titers reach the level present in the serum at the time of this interruption. We propose that the effect of autotomy before a critical period is to reinitiate a normal proecdysis. The same pattern of events occurs following autotomy of partial regenerates of crabs without eyestalks, suggesting that the decrease of serum ecdysones is brought about by some mechanism other than changes in the titer of the molt inhibitory hormone.     

Evidence for dopaminergic and opioid involvement in the regulation of locomotor activity in the land crab Gecarcinus lateralis

1. Computerized analysis of the crabs locomotor behavior revealed an initial increase in activity followed by a gradual decrease over a 12 min observation period. 2. Dopamine, in a dose-dependent manner, inhibits locomotor activity. The effect can be antagonized with the dopamine antagonist, haloperidol. This suggests that dopaminergic influences are involved with locomotor mechanisms. 3. FK 33,824, a stable opioid analog, significantly enhances the initial excitatory locomotor activity. Naloxone, a potent opiate antagonist, can block the excitatory action induced by FK 33 824. This suggests the presence of an opioid modulation mechanism in the regulation of locomotor activity. 4. Concomitant administration of the various agents results in the behavioral characteristics of the agonist appearing when the appropriate antagonist is not present. Thus, administration of dopamine + FK 33,824 + haloperidol results in enhanced locomotor activity. 5. Concomitant dopamine and FK 33,824 administration results in enhanced locomotor activity. This suggests that the opioid mechanism is closer to the last step in affecting the organism's locomotion or in initiating activity.     

Some observations on the fine structure of the sinus gland of a land crab,Gecarcinus lateralis

The dilated axon endings of the sinus glands of the brachyuran crab, Gecarcinus lateralis, are filled with homogeneously dense granules, each granule being bounded by a delicate membrane. The granules are of two orders of magnitude: 0.05 to 0.1 micro and 0.15 to 0.2 micro in diameter. Each axon ending contains granules of a nearly uniform size. Endings with granules of the larger size range predominate. Non-nervous cells endogenous to the sinus gland are scattered among the nerve endings. The cell contours are irregular, and cytoplasmic processes ramify between endings. The axons are unmyelinated, having only thin limiting membranes, and they possess many neurofibrils. Granules in preterminal portions of the axons tend to lie at the periphery of the fiber, and in some cases in chains at the core of the fiber. The granules appear to be storage and release centers for neurosecretory substances or their precursors.     

Actin-encoding cDNAs and gene expression during the intermolt cycle of the Bermuda land crab Gecarcinus lateralis

Two actin-encoding cDNAs (act1 and act2) from Gecarcinus lateralis have been sequenced or partially sequenced and the corresponding proteins deduced. The actl cDNA has a complete ORF; the act2 cDNA lacks most of the 5′ end of the coding region. The nucleotide (nt) sequences of both clones are very similar to act sequences of many organisms, the most closely related being from another arthropod, the silkmoth Bombyx mori. The proteins Actl and Act2 are more similar to vertebrate cytoplasmic actin isoforms (β-actins) than to vertebrate muscle actins (α-actins); they are also more similar to animal actins than to those of fungi or plants. Codon usage is strongly biased toward C or G in the third position. The deduced number of amino acid (aa) residues and calculated M_r for Actl are 376 aa and 41.94 kDa, respectively. The deduced aa sequence of Actl is very similar to those of muscle actins of B. mori and Drosophila melanogaster. Southern blots indicated seven to eleven act genes in the crab genome. Northern blots probed with a segment from the 3′ UTR of actl showed a single band of approx. 1.6 kb in poly(A)⁺mRNAs from epidermis, limb bud or claw muscle and in total RNAs from ovary and gill, and two bands of approx. 1.6 and 1.8 kb in total RNA from midgut gland. Western blots of one-dimensional gels of proteins from the four layers of the exoskeleton, epidermis, limb buds and claw muscle were probed with a monoclonal Ab against chicken gizzard actin; tissue- and stage-specific changes in actin content were observed. The presence of several isoforms, and differences in their number and occurrence at various stages of the intermolt cycle, were detected on Western blots of two-dimensional gels.     

Postdispersal Fruit and Seed Removal by the Crab Gecarcinus lateralis in a Coastal Forest in Veracruz,Mexico1

In the coastal semideciduous forest of La Mancha, Veracruz (Mexico), we evaluated the removal of fallen fruits and seeds of different species by the land crab Gecarcinus lateralis under different environmental conditions (age soil of forest stand, topography, and cover). Crabs removed 67 percent of the propagules in the experiments (3106 of a total of 4608), and seeds were removed in greater quantities (61.1%) than fruits (38.9%). Fruits of Maclura tinctoria (Moraceae) and Ehretia tinifolia (Boraginacea) and seeds of E. tinifolia were the most affected species. Crabs removed higher quantities of fruits in old‐soil forest and in the understory than in young‐soil forest and gaps. In experiments with seeds, only for the older forest, the crabs removed more seeds in slopes and peaks than in valleys and in the understory than gaps, whereas in the young‐soil forest, the differences were not significant. Our results suggest that G. lateralis, like Gecarcoidea natalis and Gecarcinus quadratus, is an efficient propagule predator and can exert pressure in the establishment of plant species, but in La Mancha, this also depends on site conditions and identity of the species.     

Some Observations on the Fine Structure of the Sinus Gland of a Land Crab, Gecarcinus lateralis

The dilated axon endings of the sinus glands of the brachyuran crab, Gecarcinus lateralis, are filled with homogeneously dense granules, each granule being bounded by a delicate membrane. The granules are of two orders of magnitude: 0.05 to 0.1 µ and 0.15 to 0.2 µ in diameter. Each axon ending contains granules of a nearly uniform size. Endings with granules of the larger size range predominate. Non-nervous cells endogenous to the sinus gland are scattered among the nerve endings. The cell contours are irregular, and cytoplasmic processes ramify between endings. The axons are unmyelinated, having only thin limiting membranes, and they possess many neurofibrils. Granules in preterminal portions of the axons tend to lie at the periphery of the fiber, and in some cases in chains at the core of the fiber. The granules appear to be storage and release centers for neurosecretory substances or their precursors.