U. Muglia

Ontogeny of the digestive tract in sharpsnout sea bream Diplodus puntazzo (Cetti, 1777).

The ontogeny of the digestive tract was studied histologically and histochemically in sharpsnout sea bream Diplodus puntazzo from hatching (0 DAH, Days After Hatching) until day 57 (57 DAH). At hatching, the digestive tract appeared as a histologically undifferentiated straight tube lying dorsally to the yolk sac. When the mouth opened at 3 DAH, the digestive tract was differentiated into buccopharynx, oesophagus, incipient stomach and intestine. The pancreas, liver and gall bladder were also differentiated at this stage and both the bile and pancreatic duct had opened into the anterior intestine. Active feeding began in 50% of larvae at 4 DAH, although permanence of yolk reserves until 7 DAH suggests a period of both endogenous and exogenous feeding. Nutrient absorption was first visible from 5 DAH, as colourless supra- and infranuclear vacuoles in the anterior intestinal mucosa, suggesting a lipid content, as well as supranuclear, eosinophilic vacuoles, containing protein, in the posterior intestinal mucosa. Early caecal development could be detected from 10 DAH, whereas gastric glands appeared at 30 DAH, indicating the transition from larval to juvenile stage and the acquisition of an adult mode of digestion. Goblet cells appeared in the digestive tract of sharpsnout sea bream larvae shortly after first feeding. The mucus content of goblet cells varied with the digestive region and, in the buccal cavity and oesophagus, also with the developmental phase. This study provides knowledge for better husbandry practices in the aquaculture industry, as well as for the implementation of future nutritional studies.

Three-dimensional architecture of the human myosalpinx isthmus. Scanning electron microscopy after NaOH digestion and ultrasonic microdissection.

SummaryThe three-dimensional architecture of the human isthmic myosalpinx is directly visualized by means of scanning electron microscopy after removal of interstitial connective tissue through NaOH maceration and ultrasound microdissection. These investigations show that the myosalpinx is composed of irregularly running bundles of smooth muscle cells, changing their orientation within the myosalpinx and displaying longitudinal, oblique and circular directions. The muscular bundles anastomose and intermingle with other bundles running at different levels in the oviduct wall, and actually give rise to a wide and complex muscular network in which no distinct layers are readily discernible. These morphological data are consistent with the physiological findings that the transport of gametes and embryo in very early stages in the isthmic portion of the oviduct tube is the result of a discontinuous pattern of forward and backward movements.

Cholecystokinin in White Sea Bream: Molecular Cloning, Regional Expression, and Immunohistochemical Localization in the Gut after Feeding and Fasting

Background The peptide hormone cholecystokinin (CCK), secreted by the midgut, plays a key role in digestive physiology of vertebrates including teleosts, by stimulating pancreatic secretion, gut motility, and gallbladder contraction, as well as by delaying gastric emptying. Moreover, CCK is involved in the regulation of food intake and satiation. Secretion of CCK by the hindgut is controversial, and its biological activity remains to be elucidated. The present paper addresses the regional distribution of intestinal CCK in the white sea bream, Diplodus sargus, as well as the possible involvement of hindgut CCK in digestive processes. Methodology/Principal Findings Full-lengths mRNAs encoding two CCK isoforms (CCK-1 and CCK-2) were sequenced and phylogenetically analyzed. CCK gene and protein expression levels in the different gut segments were measured 3 h and 72 h after feeding, by quantitative real-time RT-PCR and Western blot, respectively. Moreover, endocrine CCK cells were immunoistochemically detected. Fasting induced a significant decrease in CCK-2 in all intestinal segments, including the hindgut. On the other hand, no significant difference was induced by fasting on hindgut CCK-1. Conclusions/Significance The results demonstrated two CCK isoforms in the hindgut of D.sargus, one of which (CCK-2) may be involved in the feedback control of uncompleted digestive processes. On the other hand, a functional role alternative to regulation of digestive processes may be inferred for D.sargus CCK-1, since its expression was unaffected by feeding or fasting.

Ontogeny and distribution of cholecystokinin-immuno reactive cells in the digestive tract of sharpsnout sea bream, Diplodus puntazzo (Cetti, 1777), during larval development

The appearance and regional distribution of cholecystokinin-immuno reactive cells (CCK-IR) in the developing gut of larval Diplodus puntazzo were studied by means of immunohistochemistry, with the aim of understanding the role of this peptide hormone in the acquisition of digestive capacity. Immunohistochemical reaction showed CCK-IR cells from 10 days after hatching (DAH), near the pyloric sphincter and past the first bend in the midgut, as well as in the hindgut. At 25 DAH CCK-IR cells were scattered throughout the midgut, as well as in the hindgut. Since gastric glands appeared at 30 DAH, CCK-IR cells were most abundant in the anterior midgut, near and including the pyloric caeca, and just afore the ileo-rectal sphincter in the posterior midgut, as well as in the hindgut. In older larvae (39 DAH), CCK-IR cells were mainly distributed in the anterior midgut, including the pyloric caeca, as well as in the hindgut. No CCK-IR cells were detected in the foregut at any stage. The distribution pattern of CCK-IR cells differed from other species which also possess a rotated gut as D. puntazzo. In fact, although cells were abundant in regions where the ingested food is retained, so that they can be stimulated to modulating the release of digestive enzymes, a large number of cells occurred also in the hindgut.

Cholecystokinin: how many functions? Observations in seabreams.

A short overview on the regional distribution of the gastro-intestinal peptide hormone cholecystokin (CCK) in fish is presented. In particular, the results of molecular and immunological studies on seabreams, Diplodus puntazzo and Diplodus sargus, are reported, which, by demonstrating CCK in the hindgut, open new questions regarding the functional role of this hormone in that part of the intestine. The putative involvement of hindgut CCK in the feedback control of digestive processes was tested by measuring CCK gene and protein expression in fed and fasted fish. The results of this study led to hypothesize different roles for the two CCK isoforms in D. sargus, one of which related to regulation of digestive processes from pyloric caeca through hindgut. On the other hand, a functional role alternative to regulation of digestive processes may be inferred for the other isoform.

A Histological Study of Ovarian Development in the Giant Red Shrimp Aristaeomorpha foliacea (Crustacea: Decapoda: Aristeidae) from the Southern Tyrrhenian Sea (Western Mediterranean)

The reproductive features of the giant red shrimp, Aristaeomorpha foliacea, were investigated in the southern Tyrrhenian sea by experimental trawl sampling. The annual length-frequency distribution showed a multimodal trend in females, ranging between 16 and 67 mm carapace length (CL), and a unimodal trend in males (18–45 mm CL). Mature males occurred in different proportions all year round, while females displayed seasonal maturity (June—September), with a peak in July. Six oocyte developmental stages were identified, the most advanced of which (Pv, postvitellogenic) had never been described before in this species. Ovary development followed a group-synchronous pattern, with the yolked oocyte stock clearly separated from the reservoir of unyolked oocytes, suggesting that A. foliacea is a total spawner, with determinate fecundity. Based upon histological findings, a revision of macroscopic maturity staging employed in Mediterranean bottom trawl surveys (MEDITS) is proposed.

Comparative Ontogeny of the Digestive Tract in Sharpsnout Sea Bream Diplodus puntazzo Cetti and Common Pandora Pagellus erythrinus L.

The ontogeny of the alimentary tract and its associated structures (liver, pancreas, gall bladder) was compared between sharpsnout sea bream Diplodus puntazzo and Pagellus erythrinus and some differences were highlighted. The first goblet cells appeared earlier in sharpsnout seabream (5-7 DAH) and later in common pandora (9-11 DAH), as well as taste buds which appeared at 11 DAH in sharpsnout seabream and at 17 DAH in common pandora. On the contrary, stomach segmentation in a cardiac, fundic and pyloric portions was more precocious in common pandora than in sharpsnout seabream, although gastric glands were formed almost simultaneously. Early caecal development was much more precocious in sharpsnout seabream (10 DAH) than in common pandora (22 DAH). At 3 DAH in sharpsnout seabream and later (10 DAH) in common pandora, primordial mucosal folding of intestine began, which appeared well developed in the anterior portion, by 22 DAH in common pandora and by 28 DAH in sharpsnout seabream. The functional meanings of these aspects were discussed comparatively.

Architecture of the Myosalpinx of the Isthmus in the Guinea Pig by Means of Scanning Electron Microscopy

The three-dimensional architecture of the isthmic myosalpinx in the guinea pig was investigated by means of a technique that involves stretching the myosalpinx under a dissecting microscope, followed by selective digestion of the interstitial connective tissue with KOH and, finally, observation under a scanning electron microscope. The images obtained in this way prove that the myosalpinx is arranged in single bundles of smooth muscle cells which essentially do not show any specific geometrical course. Furthermore, smooth muscle cell bundles following an oblique course, not referred in the literature prior to this report, were demonstrated. These new three-dimensional microanatomical results call for a reevaluation of the functional role of oviduct motility.

Digestive enzymatic activity during ontogenetic development in zebrafish (Danio rerio)

Despite the growing importance of zebrafish (Danio rerio) as an experimental model in biomedical research, some aspect of physiological and related morphological age dependent changes in digestive system during larval development are still unknown. In this paper, a biochemical and morphological study of the digestive tract of zebrafish was undertaken to record the functional changes occurring in this species during its ontogenetic development, particularly from 24 hr to 47 days post fertilization (dpf). Endo- and exo-proteases, as well as α-amylase enzymes, were quantified in zebrafish larvae before first feeding (7 dpf). The most morphologically significant events during the ontogenesis of the gut occurred between 3 dpf (mouth opening) and 7 dpf (end of exocrine pancreas differentiation). The presence of a wide range of digestive enzymes, already active at earlier zebrafish larval stages, closely related with the omnivorous diet of this species. Increasing enzyme activities were found with increasing age, probably in relation with intestinal mucosa folding and consequent absorption surface increase. J. Exp. Zool. (Mol. Dev. Evol.) 324B: 699-706, 2015.

Three-dimensional architecture of the myosalpinx in the mare as revealed by scanning electron microscopy

The three‐dimensional architecture of the myosalpinx in the mare was investigated by means of scanning electron microscopy (SEM) after removal of interstitial connective tissue with NaOH digestion. In the extramural portion of the tubo‐uterine junction (TUJ), isthmus, and ampulla, the myosalpinx architecture is represented by a unique muscular structure which runs from the mesosalpinx to the base of the inner mucous folds. This unique muscular structure consists mainly of bundles of muscular fibers independent of one another, which show a multiple spatial arrangement and form a complex network. Such a muscular architecture is likely more suitable for stirring rather than pushing the embryos and gametes through the Fallopian tube. Anat Rec 267:235–241, 2002.