Brain Mass and Encephalization Quotients in the Domestic Industrial Pig (Sus scrofa)

open6siIn the present study we examined the brain of fetal, newborn, and adult pigs raised for meat production. The fresh and formalin-fixed weights of the brain have been recorded and used, together with body weight, to calculate the Encephalization Quotient (EQ). The weight of the cerebellum has been used to calculate the Cerebellar Quotient (CQ). The results have been discussed together with analogue data obtained in other terrestrial Cetartiodactyla (including the domestic bovine, sheep, goat, and camel), domesticated Carnivora, Proboscidata, and Primates. Our study, based on a relatively large experimental series, corrects former observations present in the literature based on smaller samples, and emphasizes that the domestic pig has a small brain relative to its body size (EQ = 0.38 for adults), possibly due to factors linked to the necessity of meat production and improved body weight. Comparison with other terrestrial Cetartiodactyla indicates a similar trend for all domesticated species.openMinervini, Serena; Accogli, Gianluca; Pirone, Andrea; Graïc, Jean-Marie; Cozzi, Bruno; Desantis, SalvatoreMinervini, Serena; Accogli, Gianluca; Pirone, Andrea; Graic, JEAN-MARIE; Cozzi, Bruno; Desantis, Salvator

Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods

The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. Sperm glycocalyx plays an important role in reproduction as it is the first interface between sperm and environment. Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods

Ultrastructural study of cultured ovine bone marrow-derived mesenchymal stromal cells.

Ovine bone marrow-derived mesenchymal stromal cells (oBM-MSCs) represent a good animal model for cell-based therapy and tissue engineering. Despite their use as a new therapeutic tool for several clinical applications, the morphological features of oBM-MSCs are yet unknown. Therefore, in this study the ultrastructural phenotype of these cells was analysed by transmission electron microscopy (TEM). The oBM-MSCs were isolated from the iliac crest and cultured until they reached near-confluence. After trypsinization, they were processed to investigate their ultrastructural features as well as specific surface marker proteins by flow cytometry and immunogold electron microscopy. Flow cytometry displayed that all oBM-MSCs lacked expression of CD31, CD34, CD45, HLA-DR whereas they expressed CD44, CD58, HLAI and a minor subset of the cell population (12%) exhibited CD90. TEM revealed the presence of two morphologically distinct cell types: cuboidal electron-lucent cells and spindle-shaped electron-dense cells, both expressing the CD90 antigen. Most of the electron-lucent cells showed glycogen aggregates, dilated cisternae of RER, moderately developed Golgi complex, and secretory activity. The electron-dense cell type was constituted by two different cell-populations: type A cells with numerous endosomes, dense bodies, rod-shaped mitochondria and filopodia; type B cells with elongated mitochondria, thin pseudopodia and cytoplasmic connectivity with electron-lucent cells. These morphological findings could provide a useful support to identify “in situ” the cellular components involved in the cell-therapy when cultured oBM-MSCs are injected

Differential Expression and Localization of Glycosidic Residues in In Vitro and In Vivo Matured Cumulus-Oocyte Complexes in Equine and Porcine Species

Glycoprotein oligosaccharides play major roles during reproduction, yet their function in gamete interactions is not fully elucidated. Identification and comparison of the glycan pattern in cumulus-oocyte complexes (COCs) from species with different efficiencies of in vitro spermatozoa penetration through the zona pellucida (ZP) could help clarify how oligosaccharides affect gamete interactions. We compared the expression and localization of 12 glycosidic residues in equine and porcine in vitro-matured (IVM) and preovulatory COCs by means of lectin histochemistry. The COCs glycan pattern differed between animals and COC source (IVM versus preovulatory). Among the 12 carbohydrate residues investigated, the IVM COCs from these two species shared: (a) sialo- and bN-acetylgalactosamine (GalNAc)-termi- nating glycans in the ZP; (b) sialylated and fucosylated glycans in cumulus cells; and (c) GalNAc and N-acetylglucosamine (GlcNAc) glycans in the ooplasm. Differences in the preovulatory COCs of the two species included: (a) sialoglycans and GlcNAc terminating glycans in the equine ZP versus terminal GalNAc and internal GlcNAc in the porcine ZP; (b) terminal galactosides in equine cumulus cells versus terminal GlcNAc and fucose in porcine cohorts; and (c) fucose in the mare ooplasm versus lactosamine and internal GlcNAc in porcine oocyte cytoplasm. Furthermore, equine and porcine cumulus cells and oocytes contributed differently to the synthesis of ZP glycoproteins. These results could be attributed to the different in vitro fertilization efficiencies between these two divergent, large-animal models

Loss of Neuron Navigator 2 Impairs Brain and Cerebellar Development

Acknowledgements We thank the study family for their enthusiastic participationPeer reviewedPublisher PD

Altered lectin-binding pattern of efferent ducts in a tricolor cat affected by Klinefelter sindrome

Recent investigations1 indicate that cat efferent ducts (EDs) play a role in reabsorption of the fluid and proteins leaving the testes, dependent also on androgen2. The males with Klinefelter syndrome (XXY) show a variable degree of androgen deficit responsible for testicular disfunction3. Since we demonstrated synthesis and secretion of glycoconjugates in normal cat EDs1, here we investigated the glycoprotein pattern of the EDs from a tricolor cat with Klinefelter syndrome (39,XXY), by means of the lectin histochemistry, utilizing a panel of 12 lectins in association with sialidase (s) treatment. Cilia of ciliated cells reacted with HPA, SBA, Con A, KOH-s-WGA, GSA II in normal cats and with MAL II, SNA, Con A and KOH-s-WGA in XXY cat. The luminal surface of non-ciliated cells bound MAL II, KOH-s-PNA, Con A in all samples, RCA120 and HPA only in normal subjects and PNA in XXY cat. The supra-nuclear cytoplasm of non-ciliated cells expressed SNA and Con A affinity in XXY cat and also MAL II, KOH-s-PNA, RCA120, SBA in normal cats. These results indicate that negative charges are mainly expressed on the cilia of XXY cat EDs, whereas a more complex glycoconjugate pattern, probably related to an more effective endocytotic apparatus, is expressed in the supra-nuclear cytoplasm of non-ciliated cells from normal EDs. References 1. Arrighi S et al. Histol. Histopathol. 2001, 25:433-44. 2. Jones RC. The epididymis: From molecule to clinical practice. Robaire and Hinton eds, Kluwer Academic/Plenum Publ. 2002,11-33. 3. Wikström AM and Dunkel L. Horm. Res. 2008, 69:317-26

Diversity of glycoconjugate pattern in the bladder urothelium of horse and donkey revealed by lectin histochemistry

The glycoconjugates of the mucosal surface of urinary bladder act as a barrier against invasion by pathogenic microorganisms and injury from toxic substances. Furthermore, they serve as a source of soluble urinary glycoproteins, which actively modify the urine composition. Glycoconjugate pattern characterizes distinct cellular populations, cell differentiation and maturation, as well as cell morpho-functional changes. We investigated the glycan pattern in the bladder epithelium by lectin histochemistry, comparatively in the horse and donkey. Tissue fragments were fixed in 4% (w/v) neutral formalin and then embedded in paraffin wax. Sections were stained with a panel of twelve lectins, in combination with sialidase (s) digestion. The entire urothelium reacted with SNA, s-PNA, s-SBA, Con A, GSA II in horse bladder, and also with MAL II and GSA I-B4 in the donkey one. The urothelium luminal surface bound MAL II, DBA, LTA in the horse and DBA and LTA in the donkey. In addition, the horse bladder stained with PNA, SBA and GSA I-B4 in the basal and luminal regions and with UEA I in the adluminal zone. These results demonstrate remarkable inter-specific difference of the glycoprotein pattern in the bladder urothelium of two Equine species, despite their close taxonomic vicinity. It is noteworthy that glycans binding PNA and UEA I lack in the epithelium lining the donkey urinary bladder