Histological, fine-structural and histochemical differences in the testicular glands of gobiid and blenniid fishes

The testicular gland (t.g.) is a glandular tissue situated adjacent to the testis of blenniid and several gobiid species. In the present study the t.g. of Blennius pavo Risso and Gobius niger L. were compared by histological and histochemical methods. In B. pavo the spermatozoa have to cross the t.g. to reach the vas deferens and thus they come into contact with the gland cells, whereas in G. niger the vas deferens is situated between the testis and the t.g. The fine structure and histo-chemistry of the t.g. cells reveal that in B.pavo the cells of the t.g. have exocrine as well as endocrine functions. The t.g. cells of B. pavo contain large amounts of lipids, form a secretion containing acid mucopolysaccharides, show positive reaction for acid phosphatase, and some cells stain for 3β-HSD and G6PD. The function of the t.g. of G. niger is exclusively endocrine. Characteristics of the gland cells of this species are well developed smooth ER and tubulovesicular or paracristal-line mitochondria. The stainings for 3β-HSD, G6PD and UDPGD give strong positive results in the whole t.g., indicating the presence of steroids and steroid glucuronides.     

Morphologie des Gebisses und Nahrungspraferenzen von dreizehn adriatischen Blenniiden (Teleostei, Perciformes)

The morphology and arrangement of the teeth in 13 sympatric species of the genus Blennius were investigated by SEM. All the species were collected in the upper littoral of the Northern Adriatic near Rovinj (Yugoslavia) and represent the total species inventory of the genus Blennius in this area. After analysing the content and length of the intestine, four groups with different feeding habits could be distinguished to which the morphological adaptations of the teeth could be correlated. There is a non-selective omnivorous group of Aufwuchs-grazer which take the inhabiting vagilfauna {Blennius gattorugine, B. incognitas, B. tentacularis ), a herbivorous group ( B. sanguinolentus, B. sphynx, B. canevae ), a carnivorous group ( B. trigloides, B. dalmatinus, B. rouxi ), and a linking group between omnivorous grazers and carnivores (B. adriaticus, B. pavo, B. nigriceps). B. galerita shows a preponderance of comblike teeth and feeds exclusively scraping microscopic algae from the rocky substrate. The morphology and position of the caniniforms vary greatly within the investigated species. The dentition on the vomer is lacking in five species and seems of no consequence in the uptake of food.     

Comparative morphology of male reproductive systems in Mediterranean blennies (Blenniidae)

The male reproductive organs of 16 species of Mediterranean Blenniidae ( Aidablennius sphynx, Blennius ocellaris, Coryphoblennius galerita, Lipophrys adriaticus, L. canevae, L. dalmatinus, L. nigriceps, Parablennius gattorugine, P. incognitus, P. sanguinolentus, P. rouxi, P. tentacularis, P. zvonimiri, Paralipophrys trigloides, Salaria pavo and Scartella cristata ) consist of pairs of testes, testicular glands, spermatic ducts, and blind pouches. Three main types of accessory sex organs were found by comparing the external morphology of the male gonads. Differences between species were observed in the volume of the testicular gland in relation to the volume of the testis and in the size and length of the spermatic ducts, and blind pouches. The anatomy of the testicular glands of all species investigated do not differ. Each gland consists of ducts that appear to be tubules which terminate at the testis periphery on one side and at the spermatic duct on the other side. Contrary to previous claims, A. sphynx has no fat body in place of the testicular gland; the gland of this species was not distinguishable from that of the other species investigated. In the Lipophrys species, in P. trigloides , and in B. ocellaris , a transition zone between testis and testicular gland is present. The testicular blind pouches empty into the spermatic ducts, into the ureter, or separately on the genital papilla. In most species, the epithelium has no or low folds, while in S. pavo it possesses high folds that nearly fill the lumen of the blind pouches. The morphological results are discussed in connection with taxonomy, ecology, and behaviour of the fishes.     

Functions of the testicular gland in two blenniid fishes, Salaria (=Blennius) pavo and Lipophrys (= Blennius) dalmatinus (Blenniidae, Teleostei) as revealed by electron microscopy and enzyme histochemistry

The functions of the testicular gland in two different blenniid fishes, Salaria pavo and Lipophrys dulmutinus , are described by fine structural and enzyme histochemical methods. In the testes of the two fishes no mature spermatozoa are found. Sperrniogenesis occurs only until the spermatidal stage. Spermatids are released into the testicular gland. During the spawning period the testicular gland functions in differentiation of spermatids, nutrition of spermatids, and secretion of sialomucins. After spawning, the testicular gland has phagocytotic functions in resorbtion of remaining spermatids, which are transformed into lipids in the gland cells. During the interspawning period the testicular gland is a storage reservoir for lipids and phospholipids which are re-transformed into testicular gland secretion in the next reproductive season. Testicular gland cells themselves do not have steroidogenetic functions, but steroids are synthesized by interstitial cells homologous to Leydig cells in other fish. Possible explanations for the reduction of the testis in L. dalmatinus and implications of the testicular gland in taking over testicular functions are discussed.     

Chromosome evolution involving Robertsonian rearrangements in Xyrichthys fish (Labridae, Perciformes)

Three Xyrichthys fish (Labridae, Perciformes), X. pavo, X. dea, and X. twistii, were cytogenetically studied. X. pavo and X. dea had 2n = 44 chromosomes, which were all acrocentric. X. twistii had 2n = 22 chromosomes consisting of eighteen meta- and submetacentric and four acrocentric chromosomes. The cellular DNA contents of X. pavo and X. twistii measured using flow cytometry were nearly equal. These results suggest that the karyotype of X. twistii evolved by decreasing the number of chromosomes by fusion events, probably Robertsonian fusion. Cytogenetic relationships among the three species were surmized on the basis of features on the karyotypes and the NOR locations. A large gap in the chromosome number between 2n = 44 and 2n = 22 is an interesting feature related to the process of chromosome evolution.     

Observations on feeding, growth and electric discharge of newborn Torpedo ocellata (chondrichthyes, batoidei)

Newly born and neonate Torpedo ocellata were obtained from gravid females caught during the autumn off the Mediterranean coast of Israel. The young torpedos, which weighed about 11 g, were fed with live Blennius pavo Risso, and doubled their weight in about 4 months. The behaviour of the torpedo during feeding was examined and photographed. When attacking a fish, the torpedo emerges from being buried in the sand and assaults the approaching prey. First, the prey is trapped under the torpedo and then directed towards the ray's mouth by its body movements. Newborn and neonate torpedos are able to discharge their electric organ. The amplitude of the discharge of one day-old fish is approximately 4 V. It increases dramatically during the first three weeks of life to 20 V, reaching an asymptotic level of about 26 V, by the end of the fourth month.     

Change of thiamin-binding protein and thiamin levels during seed maturation and germination in sesame

Thiamin-binding proteins (TBPs) occur in many types of plant seeds. The biochemical and structural properties such as subunit structure and affinity for thiamin of the proteins have been characterized. However, the change of TBP and thiamin during seed maturation and germination is little known. Sesame (Sesamum indicum L.) seeds have unique albumin TBPs, because the other TBPs from plant seeds are generally globulins. In this study, we studied the change of the TBP and thiamin levels in sesame seeds. The protein content and thiamin-binding activity of the seeds increased with seed development after flowering. Immunological analysis using an antibody against the TBP of sesame seeds showed that the protein was accumulated in seeds during maturation. The thiamin content of the seeds increased with seed development after flowering. On the other hand, the thiamin-binding activity decreased during seed germination when TBP was degraded. The thiamin content of the seeds decreased during the germination. However, the amount of thiamin phosphate in the seeds during germination was little changed. These results suggested that thiamin was accumulated and stored as a complex with TBP in sesame seeds.     

Tightly bound to DNA proteins: possible universal substrates for intranuclear processes

Tightly bound to DNA proteins (TBPs) are a protein group that remains attached to DNA after its deproteinization by phenol, chloroform or salting-out. TBP are bound to DNA with covalent phosphotriester or non-covalent ion and hydrogen bonds. They appear to be a vast protein group involved in numerous intranuclear processes. The TBPs fraction co-purified with DNA deproteinized by mild procedures is extremely heterogeneous, tissue and species-specific. The protein fraction co-purified with DNA after harsh deproteinization procedures appears to be formed from few polypeptides common to different species and tissues. Interaction sites between DNA and TBPs depend on the physiological status of the cell. The binding sites of TBPs to DNA do not co-localize with the nuclear matrix attachment regions. We hypothesize that TBPs form a universal substrate for intranuclear processes.     

A luminescent affinity tag for proteins based on the terbium(III)-binding peptide

Genetically encoded tags attached to proteins of interest are widely exploited for proteome analysis. Here, we present Tb(3+)-binding peptides (TBPs) which can be used for both luminescent measurements and affinity purification of proteins. TBPs consist of acidic amino acid residues and tryptophan residues which serve as Tb(3+)-binding sites and sensitizers for Tb(3+) luminescence, respectively. The Tb(3+) complexes of TBPs fused to a target protein exhibited luminescence characteristic of Tb(3+) by excitation of the tryptophan residue, and fusion proteins fused to one of the TPBs were successfully isolated from Escherichia coli cell lysate by affinity chromatography with a Tb(3+)-immobilized solid support.     

11-ketotestosterone stimulates putative sex pheromone production in the male peacock blenny, Salaria pavo (Risso 1810)

Male peacock blennies (Salaria pavo) release odors from their anal glands and, possibly, from their gonads that attract reproductive females. The current study investigated the effects of 11-ketotestosterone (KT) on development of the anal glands, testes, and accessory testicular organs as well as the subsequent olfactory potency of their secretions. After 3 wk of KT treatment (5 mg/kg in silastic implants), clusters of cells secreting neutral mucins differentiated in the anal gland of all treated males, whereas this occurred in only one control male. Secretions by anal glands from KT-treated males elicited greater olfactory responses, as assessed by electro-olfactography, than those from controls. Treatment with KT stimulated testicular gland growth and sialomucin secretion but had no clear effect on the germinal region of the testis; KT also stimulated enlargement of, and fluid secretion in, the blind pouches (paired evaginations of the spermatic ducts). Secretions by the testes and fluid in blind pouches from KT-treated males elicited greater olfactory responses than those from controls. In conclusion, KT stimulates development of the anal glands and testicular accessory organs and promotes odorant secretion, the putative multicomponent male peacock blenny pheromone.     

The role of male visual and chemical cues on the activation of female courtship behaviour in the sex-role reversed peacock blenny

Chemical cues during courtship in peacock blenny Salaria pavo did not add significantly to the response of visual stimuli and females did not court when a male was not visible. The results showed that female S. pavo courtship behaviour was elicited exclusively by male visual features and not by a multicomponent signal in different sensory modalities. The time spent by females courting the male and exhibiting the nuptial colouration during visual trials correlated with the size of the male's crest, suggesting a signalling function for this trait. Alternative hypothesis for the function of the putative pheromones released by the male's anal gland are discussed. Evidence that female sexual motivation decreases towards the end of the breeding season is also provided, which should be considered in future mate choice experiments.     

The adrenal gland and progesterone stimulates testicular steroidogenesis in the rat in vivo

Administration of pharmacological doses of glucocorticoid to male rats in vivo suppresses adrenal steroidogenesis and inhibits testicular steroidogenesis by inhibiting the anterior pituitary secretion of LH. In contrast, administration of ACTH to these pharmacologically-suppressed rats stimulates the adrenal secretion of progesterone and testicular steroidogenesis. The mechanism by which ACTH increases testicular steroidogenesis is dependent on the presence of the adrenal gland and is reproduced by the administration of progesterone. The conclusion from these data is that the adrenal gland has an important role in generating external signals that modulate the hypothalamic-pituitary-gonadal axis in male rats. The adrenal secretion of glucocorticoid acts as a negative signal to testicular steroidogenesis whereas progesterone acts as a positive signal. The adrenal secretion of progesterone and its conversion to testosterone by steroidogenic enzymes in the cytoplasm of the Leydig cell may provide an alternative pathway for testosterone biosynthesis and may account for the increased plasma testosterone levels during the acute phase of stress and mating.     

Endocrine control of sexual behavior in sneaker males of the peacock blenny Salaria pavo: effects of castration, aromatase inhibition, testosterone and estradiol

The effects of castration and sex steroid manipulations on the expression of sexual behavior were investigated in a small fish, the peacock blenny, Salaria pavo. In this species, large males defend nests and attract females while small "sneaker" males reproduce by imitating the female morphology and courtship behavior in order to approach nests during spawning events and parasitically fertilize eggs. Sneakers switch into nest holders in their second breeding season, thus displaying both male and female-like sexual behavior during their lifetime. We tested the effects of castration and of an aromatase inhibitor (Fadrozole, F), testosterone (T) or 17beta-estradiol (E(2)) implants on the expression of male and female-like behavior in sneakers. Sneakers were either sham-operated, castrated or castrated and implanted with vehicle, F, T+F or E(2)+F. Seven days after the treatment, sneakers were placed in a tank with a nesting male, two ripe females and an available nest. Castrated fish had lower levels of circulating T and increased the time spent displaying female typical nuptial coloration. T implants had the opposite effect, inhibiting the expression of female-like behavior and coloration. E(2) implants had no significant effect on the display of sexual behavior but the frequency of aggressive displays decreased. The results agree with previous findings in sneakers of S. pavo that demonstrated an inhibition of female-like behavior by 11-ketotestosterone (11-KT). The reported increase in T and 11-KT production when sneakers change into nest holders may thus contribute to behaviorally defeminize sneakers. Contrarily, both T and E(2) failed to promote male-like behavior, suggesting that behavioral masculization during tactic switching depends on other neuroendocrine mechanisms or that the time length of the experiment was insufficient to induce male-like behavioral changes in sneakers.     

In vitro effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on ovarian, pituitary, and pineal function in pigs

The aims of the study were: (1) to examine 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and/or prolactin (PRL) effects on in vitro secretion of progesterone (P₄) and estradiol (E₂) by luteinized granulosa and theca cells from porcine preovulatory follicles; and (2) to determine the effects of TCDD on PRL, luteinizing hormone (LH), and melatonin luteal phase in pigs. We found that TCDD itself did not affect progesterone secretion, but it abolished the stimulatory effect of PRL in the follicular cells. TCDD stimulated PRL secretion during the luteal phase and inhibited during the follicular phase. Moreover, TCDD increased luteinizing hormone secretion by pituitary cells during the follicular phase. In contrast to protein and steroid hormones, melatonin secretion in vitro was not affected by TCDD. In conclusion, it was found that the pituitary-ovarian axis in pigs is sensitive to TCDD, and the dioxin exhibited a profound ability to disrupt the ovarian actions of prolactin.     

Breeding ecology and nest aggregations in a population of Salaria pavo (Pisces: Blenniidae) in an area where nest sites are very scarce

In this paper we present data on the breeding ecology and patterns of nest aggregation in a population of Salaria pavo (Pisces: Blenniidae), in an area where hard substrates are very scarce. The study site was at Ria Formosa, an extensive littoral lagoon on the south coast of Portugal. The only hard materials available are bricks, tiles, stones and debris that clam culturists use to delimit their fields. The males of S. pavo establish nests intertidally in the holes of the bricks where they guard the eggs. Data were collected at low tide inspections of these artificial ridges and underwater behavioural observations during high tide. The main results are: (i) the same male can establish sequentially more than one nest in the same breeding season; the number of males that stay within the bricks increases before the start of the breeding season and declines towards its end; (ii) there is a large excess of mature males that do not establish nests and they are significantly smaller than the nesting males, suggesting very strong competition for nest sites; (iii) the nests of several males occur in adjacent holes of the same brick, sometimes being entirely surrounded by other nests. Thus, in this population, there is virtually no defended territory around the nest, a situation not known for other blenniid species. This pattern of nest aggregation implies both strong competition among males and reduction of aggression between neighbour parental fishes. This can be explained by the extreme scarcity and spatial distribution of available nest sites.     

A new species of Plagioporus (Caudotestis) tyrrhenicus sp.n. and its 2 new parasitologic findings in Blennius pavo Risso, 1810]

In a survey of the helminth parasites of Blennius pavo Risso, 1810 three different species of digenea were collected. Plagioporus (Caudotestis) tyrrhenicus sp.n. is described, figured and compared with other species of the subgenus Plagioporus (Caudotestis) Issaitchikov, 1928. Plagioporus (Caudotestis) tyrrhenicus differs from P. (Caudotestis) azurionis Yamaguti, 1951 in the position of cirrus sac, in sucker ratio and size of the eggs; from P. (Caudotestis) sinitzini Mueller, 1934 for extent of excretory bladder and size of the eggs; from P. (Caudotestis) synagris Yamaguti, 1952 for size of the pharynx and oesophagus, extent of the vitellaria and size of the eggs. Lepocreadium album (Stossich, 1890) Stossich, 1904 is redescribed, figured and recorded for the first time in Blennius pavo. Deretrema sp. is figured and recorded for the first time in the same host.     

<Emphasis Type="Italic">AtTBP2</Emphasis> and <Emphasis Type="Italic">AtTRP2</Emphasis> in <Emphasis Type="Italic">Arabidopsis</Emphasis> encode proteins that bind plant telomeric DNA and induce DNA bending in vitro

Telomeric DNA-binding proteins (TBPs) are crucial components that regulate the structure and function of eukaryotic telomeres and are evolutionarily conserved. We have identified two homologues of AtTBP1 (for Arabidopsis thaliana telomeric DNA binding protein 1), designated as AtTBP2 and AtTRP2, which encode proteins that specifically bind to the telomeric DNA of this plant. These proteins show extensive homology with other known plant TBPs. The isolated C-terminal segments of these proteins were capable of sequence-specific binding to duplex telomeric plant DNA in vitro. DNA bending assays using the Arabidopsis TBPs revealed that AtTBP1 and AtTBP2 have DNA-bending abilities comparable to that of the human homologue hTRF1, and higher than those of AtTRP1 and AtTRP2.