Perilipin 1 and perilipin 2 protein localization and gene expression study in skeletal muscles of European cross-breed pigs with different intramuscular fat contents.

This study investigated the lipid droplet coat proteins perilipin 1 (PLIN1) and perilipin 2 (PLIN2) localization in pig skeletal muscle and their relationship with intramuscular fat (IMF) content. PLIN1 and PLIN2 proteins were immunostained in semimembranosus muscle cross-sections from two groups of samples divergent for IMF and the gene expression was quantified. PLIN1 localized in the periphery of intramuscular adipocytes, whereas PLIN2 localized within myofibers with high lipid content. The high IMF group showed higher total cross-sectional area of PLIN1-stained adipocytes compared with the low IMF group (P<0.05), while the cross-sectional area and percentage of PLIN2-positive myofibers did not differ between IMF-divergent groups. This suggested that IMF content is mainly determined by extra-myocellular lipids. At mRNA level, PLIN2 expression was higher in high IMF muscles (P<0.05). The results indicate for the first time that in pig muscle PLIN1 and PLIN2 proteins are localized in correspondence with extra and intra-myocellular lipids, respectively.

Extrinsic afferents supplying the ovine duodenum and ileum.

The study aimed at establishing the distribution of primary sensory neurons by means of retrograde tracers Diamidino Yellow (DY) and Fast Blue (FB) injected into both the sheep duodenum and ileum, respectively. Many DY-labelled cells were found in both the distal vagal ganglia (DVG) and the spinal ganglia (SG) from T9-L3; on the contrary, the majority of the FB-labelled cells were found in the SG. In the SG, a double immunofluorescence stain was used to reveal Nitric Oxide Synthase-Immunoreactivity (NOS-IR) in association with: substance P (SP), calcitonin gene-related peptide (CGRP), neurofilament 200kDa (NF) and isolectin B(4) (IB4). The labelled neurons, both DY and FB generally ranged in size from medium to large. The majority of the SG duodenal projections were NOS negative; the majority of the SG ileal afferent neurons expressed NOS-IR. Both DY and FB NOS-IR neurons often co-localized IB4, CGRP and SP, but rarely NF.

Ileal tract and Peyer's patch innervation in scrapie-free versus scrapie-affected ovines.

Ileal Peyer's patches (PPs) are involved early during sheep scrapie infection. This study qualitatively and semi-quantitatively evaluated ileal tract and PP innervation in 29 Sarda ovines of different age, PrP genotype and scrapie status. A prominent network of fibres was detected within PPs, mainly located in interfollicular lymphoid and stromal components. Intrafollicular fibres were rarely observed, with no apparent differences between scrapie-free and scrapie-affected animals, or among ovines carrying different PrP genotypes. In adult sheep, independent of their scrapie status, nerve fibres could be detected infrequently, close to the follicle-associated epithelium. Fibres were also detected within newly formed follicles and intrafollicular microgranulomas.

A tryptophan-enriched diet improves feed intake and growth performance of susceptible weanling pigs orally challenged with Escherichia coli K88.

We tested the effect of Trp addition to a standard weaning diet and oral challenge with enterotoxigenic Escherichia coli K88 (ETEC) on growth and health of piglets susceptible or nonsusceptible to the intestinal adhesion of ETEC. Sixty-four pigs weaned at 21 d of age were divided into 3 groups based on their ancestry and BW: a control group of 8 pigs fed a basal diet (B), the first challenged group of 28 pigs fed B diet (BCh), and the second challenged group of 28 pigs fed a diet with Trp (TrpCh). The Trp diet was produced by the addition of 1 g of l-Trp/kg to the basal diet. On d 5, pigs were orally challenged with 1.5 mL suspension containing 10(10) cfu ETEC/mL or placebo, and killed on d 9 or 23. Based on in vitro villus adhesion assay, the pigs (except the B group) were classified as susceptible (s(+)) or nonsusceptible (s(-)) to the intestinal ETEC adhesion. Thus, after the challenge, treatments were B, BChs(-), BChs(+), TrpChs(-), and TrpChs(+). Pigs susceptible to ETEC were 50.0% in the BChs(+) group (3 pigs lost included) and 46.4% in the TrpChs (+) group (1 pig lost included). During the first 4 d after challenge, the challenge reduced ADG (P < 0.05), and this reduction was greater in susceptible pigs (P < 0.05) than nonsusceptible ones. Tryptophan increased ADG and feed intake in susceptible pigs (P < 0.05) from challenge to d 4, but not thereafter. Tryptophan supplementation did not improve the fecal consistency and did not reduce the number of pigs positive for ETEC in feces on d 4 after the challenge. The K88-specific immunoglobulin A activity in blood serum tended to be greater in challenged pigs (P = 0.102) and was not affected by the addition of Trp. Villous height was affected by the addition of Trp and challenge in different ways, depending on the site of small intestine. The need to consider the phenotype for the adhesion of the ETEC in studies with different supply of Trp was clearly evident. When compared with practical weaning standard diets, Trp supplementation allowed susceptible pigs to partially compensate for the effects of ETEC challenge by increasing feed intake and maintaining an adequate BW growth. This is of practical importance for the formulation of diets for pigs selected for lean growth because of the presence of an association between this trait and the susceptibility to the intestinal adhesion of ETEC.

The mesencephalic trigeminal nucleus of the duck: development and apoptosis.

The normal development of the mesencephalic trigeminal nucleus (MesV) of the white Peking duck (Anas platyrhynchos) was studied from the 9th day of incubation until hatching and during adulthood. In the early days of embryonic development, neurons are present in the posterior commissure and in the mesenchymal tissue outside the leptomeninges in addition to those in the tectal commissure (TC) and in the optic tectum. Following the internucleosomal cleavage of DNA, a massive loss of neurons in the MesV starts in the 11-day embryo and continues until the 15th day of incubation. On the 16th day, the nucleus consists of a numerically larger medial division located in the TC and a smaller lateral division within the stratum griseum periventriculare as is found in the adult animal. The programmed cell death occurring in the MesV is discussed herein and correlated with the analogous apoptotic phenomena observed in the trigeminal motor nucleus.

Mesencephalic trigeminal nucleus neurons supplying the jaw closing muscles have no spinal projection: a fluorescent double-labeling study in birds and mammals.

BACKGROUND: The present study deals with the possibility that the mesencephalic trigeminal nucleus (MeV) neurons that innervate the muscle spindles of the jaw closing muscles may also have collaterals projecting to the cervical spinal cord. At the same time, we reexamine the morphology of these cells and their location within the MeV. METHODS: The fluorescent retrograde tracers Fast Blue (FB) and Diamidino Yellow dihydrochloride (DY) were injected into the jaw closing muscles and C2-C3 spinal cord segments, respectively, of duck, rat, and rabbit in one series of experiments. In a second series of animals, the targets of the tracers were reversed. RESULTS: Retrogradely double-labeled cells (FB+DY) were not found in the MeV. On the contrary, the tracer injected into the muscles retrogradely labeled only large unipolar MeV cells, whereas the tracer injected into C2-C3 spinal cord segments labeled only small multipolar cells which were intermingled with the MeV somata of muscle spindle afferents. CONCLUSIONS: These findings exclude the possibility of spinal projections via collaterals of MeV cells supplying muscle spindles of jaw closing muscles in duck, rat, and rabbit. Moreover, the retrograde double-labeling technique evidences two cellular populations within the MeV of the duck, rat, and rabbit: large unipolar neurons which are the cell bodies of primary afferents from jaw closing muscles and small multipolar cells projecting to the upper cervical spinal cord.

Localization of motoneurons innervating the extraocular muscles of the sheep by retrograde fluorescent tracers.

Retrograde transport of the fluorescent tracers Fast blue, Evans blue, Diamidino yellow dihydrochloride, and Propidium iodide was used to determine the location of the motoneurons innervating the extraocular muscles of the sheep. An extensive superposition among the motor pools of the oculomotor nucleus (ON) has been observed. In the rostral third of the ON, a considerable merging occurs between obliquus ventralis and rectus medialis motoneurons and also between rectus ventralis and rectus medialis motoneurons. In the middle third of the ON, rectus dorsalis and levator palpebrae superioris motoneurons are intermingled with each other, and also with obliquus ventralis motoneurons dorsally and with rectus medialis motoneurons ventrally. The rostral portion of the trochlear nucleus overlaps with the caudal pole of the ON. The motoneurons innervating the obliquus dorsalis muscle are mainly contralateral with few ipsilateral exceptions. The retractor bulbi muscle receive the innervation by both the abducens and accessory abducens nuclei.

High incidence of multiple-bag fiber muscle spindles in the articularis humeri muscle of the horse.

The articularis humeri (AH) muscle of the horse is a small muscle composed of histochemically identified type I and IIA extrafusal fibers and a large number of muscle spindles. A total of 150 complete spindles with both spindle poles available were examined in serial transverse sections. On the basis of myosin ATPase-staining reactions after alkaline and acid preincubations, four types of intrafusal fibers, namely, bag1, bag2, "mixed" bag, and chain fibers, were identified. A high proportion of the spindle population (62.6%) consisted of multiple-bag spindles containing three or more (up to six) bag fibers. Also one-bag-fiber spindles were observed. The one-bag-fiber spindles containing a bag2 fiber could be traced into tandem linkages. "Mixed" bag intrafusal fibers, differing in their ATPase staining profile at the two poles, were found in spindles containing also at least one bag1 and one bag2 fiber. An unusually long extracapsular tract (up to 5,500 microns) of the bag intrafusal fibers was observed.

Peripheral territory and neuropeptides of the trigeminal ganglion neurons centrally projecting through the oculomotor nerve demonstrated by fluorescent retrograde double-labeling combined with immunocytochemistry.

The peripheral territories of sheep trigeminal neurons which send their central process to the brainstem through the oculomotor nerve were investigated by the use of fluorescent tracers in double-labeling experiments. For this purpose Diamidino yellow (DY) injection into the oculomotor nerve was combined with Fast blue (FB) injection either into the extraocular muscles (EOMs), or the cornea, or the superior eyelid. Double-labeled DY + FB cells were found in the ophthalmic region of the trigeminal ganglion in addition to single-labeled DY or FB cells. The DY and DY + FB-labeled trigeminal cells were analysed immunocytochemically for their content of substance P (SP)-, calcitonin gene-related peptide (CGRP)-, and cholecystokinin-8 (CCK-8)-like. All single-labeled DY cells showed SP-, CGRP- or CCK-8-like immunoreactivity. Double-labeled DY + FB neurons innervating the EOMs were immunoreactive for each of the three peptides, whereas double-labeled neurons supplying the cornea were only CGRP-like positive. The findings suggest that, in the sheep, trigeminal neurons which send their process centrally through the oculomotor nerve supply the EOMs, the cornea, and the superior eyelid and contain neuropeptides which are usually associated with pain sensation.

Analysis of the sternotrachealis muscle fibers in some Anseriformes: histochemistry and sex differences.

Histochemical characteristics and sizes of the fibers of the sternotrachealis (ST) muscle have been investigated in some Anseriformes (mallard, Pekin duck, Muscovy duck, and goose) of both sexes. A sexual dimorphism has been shown in the muscle of the species examined. In the mallard and Pekin duck, the male ST muscle shows type IIIA fibers in addition to the type I, IIA, and IIB fibers observed also in the female. In the Muscovy duck, the male muscle has only type I and IIA fibers, whereas the female muscle presents type I fibers and both types IIA and IIB fibers. Moreover, the mean frequencies for each fiber type were significantly different between males and females. In the goose, both male and female muscles present only type I and IIA fibers. In all the species examined, the mean areas of each fiber type are significantly different between male and female, being always larger in the male muscles. The anatomical sexual dimorphism observed in the ST muscle is discussed in relation to function.

Histochemical profile of articularis humeri muscle in the horse.

In this study the histochemical fiber type analysis of the articularis humeri muscle of the horse was performed. This muscle is composed of type I and type IIA fibers. A large number of spindles has been observed and the presence of these receptors could help to understand the role played by this muscle.

Different intrafusal fiber composition of spindles in sheep and pig extraocular muscles.

Histochemical profiles of intrafusal fibers have been examined in muscle spindles of extraocular muscles of sheep and pig. Results show that in the sheep the intrafusal content presents, in addition to chain fibers, at least one bag1 and one bag2 fiber, whereas in the pig almost all the spindles are one-bag-fiber [corrected] spindles.

The fine structure of the area postrema of the sheep.

The ultrastructural features of the area postrema (AP) were investigated in the suckling lamb, weaned lamb and adult sheep. No morphological differences were observed between lambs and sheep. Unciliated ependymal cells, linked by zonulae adherentes-type junctions and gap junctions, cover the AP ventricular surface. Clusters of pyriform neurons, glial cells, and axons are present in the parenchyma. The blood vessels are surrounded by wide perivascular spaces, which present an inner and outer basal lamina. The capillaries are of the fenestrated type. Perivascular glial cells rest on the outer basal lamina of the perivascular space and form a continuous ensheathment with their cell bodies or with flattened interdigitating processes. Along adjacent perivascular glial processes gap junctions are present. From our ultrastructural observations it appears that the overall cellular morphology of AP of the sheep does not differ substantially from that of monogastric mammals.

Fiber sizes and histochemical characteristics of the rectus abdominis muscle of the rabbit under conditions of pregnancy and mechanically induced stress.

Histochemical properties and muscle fiber diameters of the rectus abdominis (RA) muscle of the female rabbit were compared with those of RA muscle of (1) pregnant females at term of pregnancy (30 days); (2) pregnant females in which the stretch on the abdominal wall natural to pregnancy was artificially prolonged with a 40-day period of intraabdominal appliance; (3) virgin females subjected to intraabdominal appliance for 30, 45, 60, and 70 days. The RA muscle of the control female rabbits is composed of 29.73% type I, 12.13% type IIA, 57.59% type IIB, and 0.54% type IIC fibers. The stimulation on RA muscle due to either normal pregnancy or implant in virgin females provoked changes in muscle fiber diameters only; with the normal pregnancy plus the subsequent period of mechanical stimulation, changes occurred not only in muscle fiber diameters, but there was also a significant increase in the percentage of type I fibers and a concomitant decrease of type IIA and IIB fibers. The fiber-type pattern in the RA muscle of male and female rabbit also was compared. The RA muscle of the male rabbit showed more type IIA and less type IIB fibers than found in that of the female. Type I fibers were larger in the female, but type IIA fibers were larger in the male. In addition, male rabbits also were subjected to 30, 45, 60, and 70 days of RA muscle stimulation by means of an intraabdominal appliance to ascertain if the influence of such stimulation upon the RA muscle differed between the two sexes. Some differences in the response of the male RA muscle were noted.

Histochemical types and sizes of fibers in the rectus abdominis muscle of guinea pig: adaptive response to pregnancy.

Effects of pregnancy stimulation upon histochemically assessed myofibrillar ATPase and muscle fiber diameters were analysed in the rectus abdominis (RA) muscle of guinea pig. Samples of the muscle were taken at 30, 40, 50, 60, and 70 days of pregnancy and compared with samples of the same muscle taken from nonpregnant guinea pigs. Changes in muscle fiber proportions were noted through the course of pregnancy. Starting from 50 days of gestation an increase in type I fibers and a decrease in type IIB fibers were noted. Increase in muscle fiber diameters was also observed in type I, IIA, and IIB fibers. In addition, the RA muscle of the male guinea pig was compared with that of the female guinea pig and showed more type IIA and less type IIB fibers and all the three fiber types were larger than those of the female.

A scanning electron microscopic study of the mesencephalic trigeminal nucleus in duck and rabbit.

The mesencephalic trigeminal nucleus (MTN) cells of both young and adult ducks as well as of rabbits were investigated by scanning electron microscope. The rabbit showed only ovoid unipolar cells, while the duck also presented polyhedral cells. Few of these latter revealed processes originating from their surface and were recognized as multipolar cells. Some differences between the MTN cell surface of young and adult ducks were noticed. Synaptic bulbs were observed on the MTN cells in both duck and rabbit.