A histological study of the accessory reproductive organs of female Loligo forbesi (Cephalopoda: Loliginidae)

In Loligo forbesi Steenstrup, the female reproductive system consists of the ovary and accessory reproductive organs which include the oviducal gland, the nidamental gland, the accessory nidamental gland and seminal receptacle. Histological studies were made on the accessory reproductive organs of female L. forbesi. The various changes observed during maturation are described and the functional significance discussed. The secretions produced by the oviducal gland and nidamental gland apparently form the egg coats. The seminal receptacle serves to store spermatozoa after mating. The function of the accessory nidamental gland is unknown.     

The ultrastructure of chondrocytes in the cartilage of Sepia officinalis and Octopus vulgaris (Mollusca, Cephalopoda)

Thin sections of cartilage from the chondrocranium of cuttle fish and octopus were examined using the transmission electron microscope. It was found that cephalopod chondrocytes differed considerably from the chondrocytes of vertebrate cartilage; in particular they possessed many long and ramifying cytoplasmic processes and had an ultrastructure typical of protein-secreting cells. They did not, however, contain secretory granules; while vesicles and rough endoplasmic reticulum cisternae seemed to open directly to the cell surface. The cell body and processes contained cytoskeletal structures: microtubules were easily recognized, but intermediate and thin filaments were difficult to make out as they were frequently clumped into bundles. Some chondrocytes contained conspicuous accumulations of hemocyanin. The cytoplasmic processes possessed intercellular contacts similar to gap junctions. Also present on processes and the cell body were cell-extracellular matrix focal adhesions. The chondrocytes were not polarized or arranged in any preferential spatial order, however, with their processes they formed a three-dimensional network throughout the cartilage tissue. Ultrastructural findings are discussed in relation to the singular morphofunctional characteristics of cephalopod cartilage which shares features with both the cartilage and bone of vertebrates.     

Morphology and electrical properties of Schwann cells around the giant axon of the squids Loligo forbesi and Loligo vulgaris.

The first successful dye-fills of Schwann cells around the split giant axon of Loligo show them to be spindle-shaped cells ca. 600 microns long and 20 microns wide lying parallel to the axonal axis. There are some 50,000 Schwann cells per cm2 of axonal membrane. Only a small part (ca. 6% of each Schwann cell membrane) is in contact with the periaxonal space, the remainder is overlain by adjacent Schwann cells, or applied to the basal lamina. The mean membrane potential of the Schwann cells in artificial seawater (ASW) varies from around -40 mV in fresh split-axon preparations to around -60 to -70 mV after 1-2 h; this hyperpolarization is not seen in preparations dissected and maintained in Ca2(+)-free ASW. Electrical- and dye-coupling (abolished by prior octanol treatment) is present between Schwann cells, but is weaker in cells with lower (less negative) membrane potentials. The implications for potassium homeostasis around the axon are briefly discussed.     

Isolation, Cloning, and Characterisation of a trp Homologue from Squid (Loligo forbesi) Photoreceptor Membranes

Abstract: The invertebrate phototransduction system is a valuable model of the ubiquitous inositol lipid signalling system. Taking advantage of the ability to obtain relatively large amounts of retinal material from the cephalopod eye, partial protein sequence data were obtained for a 92-kDa component isolated from a detergent-insensitive cytoskeletal fraction of a squid retinal microvillar membrane preparation. Degenerate oligonucleotides, designed on the basis of these sequence data, were used to isolate a full-length cDNA, encoding the 92-kDa component, using both cDNA library screening and 5'-rapid amplification of cDNA ends (5'-RACE) techniques. Comparison of the amino acid sequence encoded by this cDNA with entries in the OWL composite protein sequence database reveals greatest sequence similarity with the products of the Drosophila trp and trpl genes. Greatest variation from the Drosophila Trp protein is seen in the carboxyl-terminal region, which is considerably truncated in the squid protein and which accounts for most of the substantial difference in molecular weight seen between these proteins. This variation may be significant as the carboxyl-terminal domain has been shown to be in the regulation of several ligand-gated channels. The carboxyl-terminal domain has been expressed and shown to interact with calmodulin in a calcium-dependent fashion, thereby supporting this hypothesis. The likely occurrence of other homologues in a variety of systems suggests that this is a novel and important family of regulated ion channels involved in calcium signalling.     

Evidence for an intermediate type of spermatozoon: ultrastructural studies of spermiogenesis in the cuttlefishRossia macrosoma (Cephalopoda,Decabrachia)

Summary Testes and spermatophores of the decapod cephalopodRossia macrosoma have been investigated by electron microscopy. Cytochemical analysis, by use of the periodic acid thiosemicarbazide-silver proteinate (PA-TSC-SP) technique, allows the precise localization of glycogen within the spermatozoon. Nuclear morphogenesis occurs in three stages (termed A, B and C), resulting in the transformation of the nucleus from an ovoid shape (stage A) into an elongated form (stages B and C). Prior to elongation of the spermatid nucleus, the spheroidal proacrosome, derived from the Golgi complex, becomes located at the apex of the nucleus. Later, the proacrosome elongates and its internal contents differentiate into an apical vesicle, electron-dense rod and electron-lucent subacrosomal fossa. A longitudinal microtubular manchette is closely associated with the elongating proacrosome. At this stage, the spermatid nucleus also elongates and its associated microtubular manchette is continuous with that surrounding the proacrosome. Large quantities of glycogen granules,-particles, about 130–145 Å in diameter, occur in the midpiece in relation to both the specialized external plasma membrane of the mitochondrial sleeve and within or around the axonome in the tail piece. We compare the features of spermiogenesis and the spermatozoon ofR. macrosoma with those documented in the literature for species from a number of phyla with the so-called primitive type of spermatozoon and which practise external fertilization, and the modified type in species practising internal fertilization. We suggest that the spermatozoon and spermiogenesis ofRossia demonstrates characteristics intermediate between these two groups and that the spermatozoon should be classified as an intermediate type.     

Pattern of Inheritance of Microsatellite Loci in the Squid Loligo pealeii (Mollusca: Cephalopoda)

Six microsatellite loci are described for the squid Loligo pealeii. All loci exhibit some degree of allelic diversity. The pattern of inheritance was tested for 3 loci through an analysis of the filial genotypes from a female-male mating. At all 3 loci, the ratios of the filial genotypes conformed to the ratios expected by Mendelian inheritance. The hypervariable loci will be useful in studies on sexual selection in this species, whereas the relatively less variable loci will be useful to address questions of population structure. Received March 13, 2000; accepted May 22, 2000.     

Digestive absorption in Octopus vulgaris (Cephalopoda: Octopoda)

In Octopus , the caecum, the pancreas and the liver are involved in the absorption of ³H-glycine-labelled food. The caecum exhibits the highest specific activity while the liver absorbs the largest proportion of labelled material. The administration of stained food shows in vivo alternating phases of absorption and secretion in the liver and continuous absorption in the caecum.     

Oxygen consumption and ammonia excretion rates in Octopus maya,loligo forbesi and Lolliguncula brevis (Mollusca: Cephalopoda)

Oxygen consumption and ammonia excretion rates were investigated in young Octopus maya (hatching to 139 days old; 0.11–81.23 g wet body weight, BW; 22.5–23.9°C), young squids of Loligo forbesi (hatching to 45 days old; 9.4–115.3 mg BW; 12.3–13.1°C) and young squids of Lolliguncula brevis (2.00–39.98 g BW; 23.8–24.7°C). Except at hatching, oxygen consumption and ammonia excretion rates on an individual basis (M) of these three cephalopods increased linearly with increasing body weight (BW) expressed as M = aBW^b . Values of b for oxygen consumption were 0.900, 0.910 and 0.848 and for ammonia excretion were 0.744, 0.809 and 0.751 for O. maya, L. forbesi and L. brevis, respectively. Among the three species the value a varied widely, while b was similar for both oxygen consumption and ammonia excretion rates. Based upon these data, metabolism for hatchlings of O. maya and L. forbesi was estimated to be relatively lower than that of older juveniles. The O/N ratios for hatchlings of O. maya and L. forbesi were relatively high and indicate an apparent dependence upon lipids in the immediate post‐hatching period, followed by standard protein energy utilization thereafter.     

Drilling by Octopus vulgaris(Mollusca: Cephalopoda) in the Mediterranean

Octopus vulgaris drills holes in the shells of a variety of molluscs. The walls of the cavities drilled exhibit dissolution of mineral and organic material. The features which characterize the cavities have been described. The composition and structure of the shell itself is important in determining the size, shape and form of the cavity drilled, and not the size of the octopus. Capture, drilling the shell, and eating the occupant may take less than one hour.     

Integumental amino acid uptake in a carnivorous predator mollusc (Sepia officinalis, Cephalopoda)

The epithelial cells of the integument of body, arms and tentacles of Sepia officinalis present on their apical membrane a well-organised brush border and show the morphological and histochemical characteristics of a typical absorptive epithelium. The ability of the integument to absorb amino acids was investigated both in the arms incubated in vitro and in a purified preparation of brush border membrane vesicles (BBMV). Autoradiographic pictures of the integument after incubation of the arms in sea-water with or without sodium, showed that proline intake was Na+-dependent, whereas leucine intake appeared to be a largely cation-independent process. Time course experiments of labelled leucine, proline and lysine uptakes in BBMV evidenced that these amino acids are accumulated within the vesicles in the presence of an inwardly directed sodium gradient. The sodium-driven accumulation proves that cationic and neutral amino acids are taken up by the apical membrane of the epithelium of Sepia integument through a secondary active mechanism. For leucine, a 90% inhibition of the uptake was recorded in the presence of a large excess of the substrate. In agreement with the autoradiography results, an analysis of the cation specificity transport in BBMV showed that leucine uptake had a low cation specificity, whereas lysine and proline uptakes were Na+-dependent. An excess of lysine and proline, which share with alanine two different transport systems in the gill epithelium of marine bivalves, reduced eucine uptake. The possible role of the absorptive ability of the integument in a carnivorous mollusc is discussed.     

Adenosine triphosphatase localization in the branchial heart of Sepia officinalis L. (Cephalopoda)

Sodium- and potassium-dependent adenosine triphosphatase (Na+--K+-ATPase) is demonstrated in the branchial heart of Sepia officinalis L. by biochemical, cytochemical and autoradiographical methods. The biochemical data indicate the presence of Na+--K+-ATPase, shown by potassium and magnesium dependency and inhibition by ouabain. Cytochemically and autoradiographically, the enzyme is localized in the sarcolemma of the muscle cells. The positive reaction of the transparent cells (type I cells) is due to activity of alkaline phosphatases. The dark cells (type II cells) react negatively. In addition to the Na+--K+-ATPase, a magnesium-activated adenosine triphosphatase (Mg2+-ATPase) and a bicarbonate-stimulated ATPase (HCO3(-)-ATPase) are localized in the mitochondria.     

Serotonergic regulation of the central heart auricles of Sepia officinalis L. (Mollusca, Cephalopoda)

In pharmacological bioassays on isolated isotonically suspended auricles of Sepia officinalis, the regulatory action of the neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) on these autonomous contractile compartments was demonstrated. 5-HT induced concentration-dependent positive effects on frequency and tone, whereas the concentrations/response curve for the amplitude showed a biphasic course. All applied antagonists inhibited mainly the effect of 5-HT on frequency and amplitude. The chronotropic effects of 5-HT were blocked mainly by the 5-HT(1,2) antagonist methiothepin (pA(2)=8.01), the 5-HT(1a) antagonist NAN-190 (pA(2)-) and in lesser extent by the 5-HT(1,2) antagonist mianserin (pA(2)=6.81). In the presence of each antagonist applied the 5-HT action on amplitude was transformed into a positive inotropic effect with the maximum under influence of NAN-190 and the 5-HT(2,1c)-antagonist ketanserin. The auricular tone was also influenced by the antagonists and in combination with methiothepin it turned into strong negative tonotropic effect. In addition to the pharmacological bioassays, the presence of 5-HT in nerve endings within the auricle wall was demonstrated by immunohistochemical and fluorescence microscopic findings. Altogether the findings presented here confirm that 5-HT evokes excitatory effects on the autonomous contractile auricle of S. officinalis and acts obviously over different receptors, whereby a 5-HT(1)- and a 5-HT(2)-like seem to be involved.