The immunoglobulin A isotype of the Arabian camel (Camelus dromedarius) preserves the dualistic structure of unconventional single-domain and canonical heavy chains.

Introduction: The evolution of adaptive immunity in Camelidae resulted in the concurrent expression of classic heterotetrameric and unconventional homodimeric heavy chain-only IgG antibodies. Heavy chain-only IgG bears a single variable domain and lacks the constant heavy (CH) γ1 domain required for pairing with the light chain. It has not been reported whether this distinctive feature of IgG is also observed in the IgA isotype. Methods: Gene-specific primers were used to generate an IgA heavy chain cDNA library derived from RNA extracted from the dromedary's third eyelid where isolated lymphoid follicles and plasma cells abound at inductive and effector sites, respectively. Results: Majority of the cDNA clones revealed hallmarks of heavy chain-only antibodies, i.e. camelid-specific amino acid substitutions in framework region 1 and 2, broad length distribution of complementarity determining region 3, and the absence of the CHα1 domain. In a few clones, however, the cDNA of the canonical IgA heavy chain was amplified which included the CHα1 domain, analogous to CHγ1 domain in IgG1 subclass. Moreover, we noticed a short, proline-rich hinge, and, at the N-terminal end of the CHα3 domain, a unique, camelid-specific pentapeptide of undetermined function, designated as the inter-α region. Immunoblots using rabbit anti-camel IgA antibodies raised against CHα2 and CHα3 domains as well as the inter-α region revealed the expression of a ~52 kDa and a ~60 kDa IgA species, corresponding to unconventional and canonical IgA heavy chain, respectively, in the third eyelid, trachea, small and large intestine. In contrast, the leporine anti-CHα1 antibody detected canonical, but not unconventional IgA heavy chain, in all the examined tissues, milk, and serum, in addition to another hitherto unexplored species of ~45 kDa in milk and serum. Immunohistology using anti-CHα domain antibodies confirmed the expression of both variants of IgA heavy chains in plasma cells in the third eyelid's lacrimal gland, conjunctiva, tracheal and intestinal mucosa. Conclusion: We found that in the dromedary, the IgA isotype has expanded the immunoglobulin repertoire by co-expressing unconventional and canonical IgA heavy chains, comparable to the IgG class, thus underscoring the crucial role of heavy chain-only antibodies not only in circulation but also at the mucosal frontiers.

Developmental events in the lung of the Japanese quail (Coturnix coturnix japonica): Morphological, histochemical and electron-microscopic studies.

The lung of birds is the most complex and efficient gas exchanger in the air-breathing vertebrates. A total number of 27 normal Japanese quail embryos during the pre-hatching period were used. The current work aimed to investigate the histomorphological, histochemical and ultrastructural changes of the lung at different stages of development using light and electron microscopy. The results showed that the respiratory primordium was observed on the 2nd embryonic day (ED) as a ventral out-pouching of the primitive foregut into the surrounding mesenchyme. The first evidence of the lung buds appeared on the 3rd ED. On the 4th ED, the buds increased in size. In addition, the secondary bronchi budded from the epithelial lining of the primary bronchus into the surrounding mesenchyme. On the 6th ED, the number of the secondary bronchi increased and began to give rise small tubules (parabronchi). On the 7th ED, telocytes (TCs) were seen extensively in relation to the undifferentiated mesenchymal cells (UMC) and surrounding the developing bronchi and blood vessels. On the 8th ED, the number of the small-diameter, smooth-walled parabronchi (PR) was considerably increased. On the 9th ED, anastomosing of the developing parabronchi was observed forming large diameter ones and the open-type endocrine cells in the parabronchial wall were observed. On the 10th ED, deep costal impressions on the lung tissue were seen, the atrial primordia could be observed and the primordia of all air sac were developed. In conclusion; morphogenesis and differentiation of the developing lung epithelium depend on epithelial-mesenchyme interaction.

Pregnancy-associated glycoprotein (PAG) family: transcripts and gene amplicons in camelids.

In this study, the placental localization of PAG-like transcripts and genomic existence of PAG-like amplicons in new-world (Lp, Lama pacos, alpaca) and old-world camelids (Cb, Camelus bactrianus, bactrian; Cd, Camelus dromedarius; dromedary) are reported for the first time. Sections of Lp (150-347 days post coitum), Cd (43-90 cm crown-rump length) and Cb (term) placentas were used for heterologous (ht; cross-species) autoradiographic in situ hybridization (aISH) with single-stranded diagnostic (antisense) or control (sense) [alpha-(35)S]dATP-labeled 323 nt porcine PAG8 (pPAG8) cDNA probes produced by asymmetric PCRs. The aISH with antisense (35)S-pPAG8 probe identified camelid PAG-like (LpPAG, CbPAG and CdPAG) mRNA expression restricted to chorionic epithelium cells within placentas of camelids. In addition, genomic DNA (gDNA), isolated from placental sections were used as templates for camelid PAG-like gene amplicon production by PCR. Specificity of the obtained multiple camelid gDNA PAG-like amplicons was confirmed by double ht-Southern hybridizations with [alpha-(32)P]dATP-labeled 611 bp pPAG5 and pPAG10 double-stranded cDNA probes. The double ht-Southern hybridizations of camelid gDNA amplicons (with pPAG5 and -10 probes) allowed the identification of length-polymorphism of LpPAG, CbPAG and CdPAG genes, coding catalytically active and potentially inactive forms. Such an application of porcine PAG probes may be advantageous for future identification of still undiscovered PAG-like families in other eutherian species.

Microvascular architecture of the fetal cotyledons in water buffaloes (Bubalus bubalis) during different stages of pregnancy.

To elucidate the morphological background of physiological differences between bovine and buffalo gestation forty-two placentae ranging from the 3rd to 10th month of pregnancy were used to study the microvascular architecture of the fetal cotyledons in the buffalo. The tissues were prepared for light and scanning electron microscopy by paraformaldehyde fixation and corrosion casting of the fetal cotyledonary vascular system. Histology and vascular casts revealed the buffalo fetal cotyledons to consist of a series of conical villous trees changing from a wide to slender shape during pregnancy, and with a base strictly facing the fetal side. The branches of these trees, intermediate and terminal villi, projected radially from the stem, thus representing a rough type of villous tree. A second type of tree lacked these branches and was therefore termed smooth villus. The rough type was most prevalent, and the intermediate and terminal villi showed capillary complexes arranged in stories by the 4th to 5th month of gestation. The stories became broader and denser with the progress of pregnancy (6th to 7th month of gestation), due to extensive growth of new capillaries and simultaneous development of convolutions causing the vascular ridges of the terminal villi to appear bushy. Near term (9th to 10th month) the capillary system became very dense, particularly at the margin of the vascular ridges, leaving only narrow spaces for the corresponding maternal septal tissue. In detail, at its base the trunk of each villous tree contained a single central stem artery which originated from the allantochorionic arterial system, and 1-3 parallel peripheral stem veins. When approaching the cone tip, these vessels branched into new stem arteries and veins, each giving rise to arterioles and venules according to the principle vascularization of the stem villus first, followed by intermediate and terminal villi. The capillary complex of the terminal villi consisted of arterial capillary limbs, capillary loops with sinusoidal dilatations and anastomoses, and venous capillary limbs. The latter converged into venules of terminal and intermediate villi which joined stem veins leading into allantochorionic veins. In conclusion, the fetal vasculature of the buffalo placentome was greatly increased from early pregnancy to near term. This was denoted by the general development of stem villous trees and an increase in the volume and density of the capillary system of the terminal villi, reflecting the increasing need of materno-fetal substance exchange in the buffalo placenta, particularly near term.

Morphological characteristics of the tongue and its papillae in the donkey (Equus asinus): a light and scanning electron microscopical study.

The morphology of the donkey tongue and its papillae were investigated by macroscopy and by light and scanning electron microscopy in ten adult animals (six males and four females). The spatula-shaped tongues measured about 28 cm in length, 4.5 cm in breadth and 3.5 cm in thickness. Samples from different areas of four tongues were grossly examined and pieces were processed for light and scanning electron microscopy. Filiform papillae were distributed mainly on the dorsum of the tongue, being thin and relatively short at the apex, conical and scaly in the main part (triangular zone) of the body, and thin and longer at the caudal part of the body. Few of them were found on the lateral surfaces. Fungiform papillae appeared scattered mainly on the lateral surfaces. They were mostly rounded (about 1.0 mm in diameter), but lobulated forms were also observed. Filiform and fungiform papillae were both completely devoid of taste buds, indicating a more mechanical function. The vallate papillae were 2-3 in number, located at the most caudal part of the body. They were three to four times as large as the fungiform papillae (about 5.6 mm in diameter) each with a wide circular groove around the central bulbous projection. Secondary grooves originating from the primary one were also demonstrated. The vallate papillae contained many taste buds with taste pores opening deeply into the papillary groove. Fine filiform papillae were demonstrated on the bulb-like part of the vallate papillae. The donkey tongue had sinister and dexter well-developed sets of foliate papillae close to the basis of the palatoglossal arch. They were arranged in the form of numerous leaves separated by deep, variably wide grooves and contained a very large number of taste buds. It is believed that the existence of well developed foliate papillae in donkey may substitute the comparatively few vallate papillae in this species.