Genetic diversity and population structure in native chicken populations from myanmar, Thailand and laos by using 102 indels markers.

The genetic diversity of native chicken populations from Myanmar, Thailand, and Laos was examined by using 102 insertion and/or deletion (indels) markers. Most of the indels loci were polymorphic (71% to 96%), and the genetic variability was similar in all populations. The average observed heterozygosities (H O ) and expected heterozygosities (H E ) ranged from 0.205 to 0.263 and 0.239 to 0.381, respectively. The coefficients of genetic differentiation (Gst) for all cumulated populations was 0.125, and the Thai native chickens showed higher Gst (0.088) than Myanmar (0.041) and Laotian (0.024) populations. The pairwise Fst distances ranged from 0.144 to 0.308 among populations. A neighbor-joining (NJ) tree, using Nei's genetic distance, revealed that Thai and Laotian native chicken populations were genetically close, while Myanmar native chickens were distant from the others. The native chickens from these three countries were thought to be descended from three different origins (K = 3) from STRUCTURE analysis. Genetic admixture was observed in Thai and Laotian native chickens, while admixture was absent in Myanmar native chickens.

Morphological studies on the vascular architecture in the boar spermatic cord.

The morphological features of the testicular artery and the pampiniform plexus in the boar spermatic cord were evaluated by light microscopy, corrosion cast technique combined with scanning electron microscopy and by transmission electron microscopy. The testicular veins could be subdivided into 4 types according to their perivascular elements and their location to the testicular artery. Type I vein consisted of large veins and gave rise to type II and III veins. Type II vein was composed of a single layer of veins and ran along the testicular artery, while type III vein consisted of several layers of veins and was located between type II veins. Type IV vein, which was regarded as a venous portal system, was composed of small muscular and pericytic venules and was located in the tunica adventitia of type II vein and the testicular artery. Occasionally, type IV vein penetrated deep into the tunica media of the testicular artery, accompanied by a fenestrated endothelium in its thin portion. The direct arterio-venous anastomosis between the pampiniform plexus and the testicular artery was not found. Type IV vein was considered to be the most important vessel in reducing the capacity of the barrier between the testicular artery and the veins (type II and III). It is, therefore, suggested that type IV venous network may play a role in transferring the substances between the artery and the veins.

Ultrastructure of cardiac myocytes in the greater bandicoot rat (Bandicota indica).

Cardiac myocytes in the left ventricle and atrium of the greater bandicoot rat (Bandicota indica) were examined by transmission electron microscopy. The fine structure showed typical features of mammalian cardiac myocytes. In atrial myocytes, however, the areas occupied by mitochondria were much smaller than that filled with myofibrils. The decrease in mitochondria and the abundance of myofibrils are thought to be an ultrastructural adaptation to the large body size of this species. Many conducting myocytes were observed in both atrium and ventricle. The atrial conducting myocytes were ultrastructurally different from the Purkinje fibers of the ventricle. We suggest that the abundance and the ultrastructural variation of conducting myocytes are related to the physiological aspects, such as body size, of this animal.

Classification of the cycle of the seminiferous epithelium in the common tree shrew (Tupaia glis).

The classification of the cycle of the seminiferous epithelium was carried out in the common tree shrew (Tupaia glis). The tree shrew captured in Thailand were fixed with Bouin's fixative, embedded in paraffin wax, and stained with PAS-hematoxylin. The cycle was classified into twelve stages on the basis of the acrosomal changes of spermatids. Relative frequencies of stages form I to XII were 11.9, 7.2, 8.9, 22.5, 12.9, 9.7, 8.0, 5.9, 4.0, 3.2, 2.9, and 3.6%, respectively. Different stages did no appear in a cross-sectioned tubule as did in primates. The head of matured spermatid was discoidal in shape and different from that of primates and rodents. Spermatogenesis of the common tree shrew is different from that of primates and rodents according to its morphological features.

Morphological features of the spermatic cord in the musk shrew (Suncus murinus) with special reference to extratesticular Leydig cells.

Morphological features of the testicular artery and vein in the spermatic cord of the musk shrew (Suncus murinus) were evaluated by light microscopy, transmission electron microscopy, corrosion cast technique combined with scanning electron microscopy and immunohistochemistry. The vascular architecture in the spermatic cord of the musk shrew was simple. The testicular artery in the musk shrew was straight and accompanied by 1 to 3 branches of testicular vein. The testicular vein was also straight and anastomosed with each other in some points along its length, but it did not form a delicate pampiniform plexus. In the middle and distal portions of the spermatic cord, the tunica adventitia of the artery and vein was joined together to form a single connective tissue septum. Clusters of cells were found in this connective tissue septum in the middle portion of the cord. These cells were located close to the arterial wall and nerve endings, but they did not appear inside of neurium. They showed several typical characteristics similar to Leydig cells, and they were positive for 3beta hydroxysteroid dehydrogenase (HSD) antibody. Ultrastructural and immunohistochemical studies also indicated that the cells in cluster found in the vascular wall of the musk shrew spermatic cord may be equivalent to Leydig cells in testes. These extratesticular Leydig cells had characteristics of the active steroid-producing cell and seemed to be another source of testosterone.

Osteometrical skull character in the four species of tree shrew.

The skull size and shape were osteometrically examined in the four species of the tree shrews (Tupaia tana, T. javanica, T. minor and T. dorsalis). We suggest that the skull characters were affected by the species specific behavior and terrestrial, arbo-terrestrial and arboreal life, among the genus Tupaia. The neurocranium was laterally narrower in the braincase area, and the splanchnocranium was longer only on dorsal side in T. tana, and these characters were opposite to T. minor. The principal component analysis confirmed the obviously separated clusters among T. tana, T. javanica and T. minor, affected by the adaptation for each behavior. T. dorsalis was considered as terrestrial species from the results of proportion analysis and the principal component analysis.

Sympatric distribution of the two morphological types of the common tree shrew in Hat-Yai districts (South Thailand).

The two color types (grayish northern and reddish southern types) of the common tree shrew (Tupaia glis and Tupaia belangeri) were co-distributed in Hat-Yai region (South Thailand). Although the Isthmus of Kra in South Thailand has been considered as distribution barrier of the two types, the sympatric distribution of both types was confirmed in southern side of the Isthmus. In the principal component analysis, the skull measurement character from Hat-Yai region could also be separated into the northern and southern groups according to the skin color identification of corresponding individuals. We could generally distinguish the common tree shrew into two types by skull morphology as well as external skin color.

Immunohistochemical survey of the gut endocrine cells in the common tree shrew (Tupaia belangeri).

Regional distribution and relative frequency of endocrine cells in the gastrointestinal tract of the common tree shrew (Tupaia belangeri) were studied immunohistochemically. Ten types of immunoreactive endocrine cells were localized in the gastric mucosa, i.e., chromogranin-, serotonin-, gastrin-, somatostatin-, bovine pancreatic polypeptide (BPP)-, enteroglucagon-, pancreatic glucagon-, peptide tyrosine tyrosine (PYY)-, motilin-, and substance P (SP)-immunoreactive (IR) cells. In the intestine, 13 types of immunoreactive cells were observed, i.e., chromogranin-, serotonin-, somatostatin-, gastrin-, BPP-, enteroglucagon-, PYY-, secretin-, cholecystokinin (CCK)-, gastric inhibitory peptide (GIP)-, motilin-, neurotensin-, and SP-IR cells. The regional distribution and relative frequency of the cell types varied along the gastrointestinal tract. Basically, the types, distribution, and relative frequency of the gut endocrine cells were similar to those reported in other mammalian species. However, some characteristic findings were noted in the present study: (1) the considerably large number of gastrin-IR cells in the pyloric region; (2) numerous serotonin-IR cells in the stomach; (3) appreciable number of BPP-IR cells in the transitional region of the stomach; and (4) wide distribution of PYY- and motilin-IR cells in the gut.

An osteometrical study of the cranium and mandible of the lesser mouse deer (Chevrotain), Tragulus javanicus.

The lesser mouse deer (Tragulus javanicus) is one of the most primitive ruminants. Skulls of lesser mouse deer were measured to evaluate their growth. The age was estimated from the eruption and attrition of molars on the mandible. A total of 95 specimens was divided into five age groups, and the growth pattern in each sex was established for some measurements. The relative growth coefficients were calculated for the head and body length and several parts of the cranium and mandible against the profile length. Sexual dimorphism could not be found on the skull in any age group except for measurements related to the upper canine. However, females showed a higher value in head and body length in the oldest group. This feature was consistent with an interpretation of mouse deer society as monogamous. Growth was more rapid in length than in width. Various statistical analyses showed that the visceral cranium related to masticatory facility was much better developed than the neurocranium. However, observations on the shape of the mandible ramus and of the premolars implied that the masticatory function was not suitable for taking roughage. This speculation agreed with previous reports on the feeding habits and digestive organs of the mouse deer.

Monoclonal antibody TSd-1 is specific to elongating and matured spermatids in testis of common tree shrew (Tupaia glis).

The monoclonal antibody (MAb), named TSd-1, specific to spermatogenic cells of the common tree shrew (Tupaia glis) was established and characterized using immunohistochemistry and immunoblotting. MAb TSd-1 reacted with elongating and matured spermatids in a stage-dependent manner. TSd-1 recognized a 94 kilodalton (kDa) peptide in the plasma membrane and cytosol. Additionally, an extremely weak 107 kDa band was detected only in the cytosol. The reactions were not detected in round spermatids. In elongating Stage VI spermatids, the plasma membrane and the granular structure within the cytoplasm were intensely positive, and most intense after the appearance of new round spermatids in the lower layer (Stage I). The reactions were observed neither in the other organs of the common tree shrew nor in the testes of other animals, indicating that TSd-1 antigen is specific to the spermatogenic cells of the common tree shrew, and may act on elongating or matured spermatids.

A lectin-histochemical study on the seminiferous epithelium of the northern smooth-tailed tree shrew (Dendrogale murina) and the Java tree shrew (Tupaia javanica).

Lectin-binding patterns in the testes of the northern smooth-tailed tree shrew, Dendrogale murina and Java tree shrew, Tupaia javanica were studied by light microscopy and compared the data with those of the common tree shrew. Four lectins (PNA, SBA, BPA and GS-II) were used in this study. Peanut (Arachis hypogaea) agglutinin (PNA), soybean (Glycine max) agglutinin (SBA) and Bauhinia purpurea agglutinin (BPA) showed a strong reaction in the acrosomal region from Golgi to acrosome-phase spermatids in three species of tree shrews. These lectins also showed a granular positive reaction in the cytoplasm from acrosome to maturation-phase spermatids in three species, except that BPA revealed no granular reaction (though it was positive) in the spermatid cytoplasm of the northern smooth-tailed tree shrew and that PNA revealed no reaction in the spermatid cytoplasm of the common tree shrew. While, Griffonia simplicifolia-II agglutinin (GS-II) showed a positive reaction in the acrosomal region of Golgi-phase spermatids in three species of tree shrews. Although GS-II was positive in the spermatocyte cytoplasm of three species, it showed granular in the northern smooth-tailed tree shrew and common tree shrew but not granular in the Java tree shrew. Thus, the lectin-binding patterns in testes were similar among three species belonging to the Order Scandentia. However, slight differences were also detected even among these phylogenetically-close species.

Evidence for a direct arteriovenous connection (A-V shunt) between the testicular artery and pampiniform plexus in the spermatic cord of the tree shrew (Tupaia glis).

In the tree shrew Tupaia glis, 5 or 6 small ramifying arterioles arose directly from the testicular artery and then gave off numerous small capillaries. The capillaries made a series of anastomoses with neighbouring counterpart capillaries to become a complicated network. Some of the capillaries drained into a small venule, which was connected directly with the testicular vein (pampiniform plexus), to form an arteriovenous connection (A-V shunt) between the testicular artery and the pampiniform plexus. This A-V shunt appears to make the transfer of substances from the pampiniform plexus to the testicular artery more efficient. In addition, the shunt may control the volume of the blood draining into the testis. The capillaries were covered by vesiculated cells which were located adjacent to the pericytes. The vesiculated cells contained abundant mitochondria, rough endoplasmic reticulum, a well developed Golgi complex and cytoplasmic vesicles. Their cellular processes were long and surrounded more than one capillary. The morphological features of the vesiculated cells suggest that they may synthesise substances that are released into the network and which affect the activity of the capillaries. Since the cellular processes contacted each other, the cells could provide support for the capillaries and prevent their collapse when the shunts are closed.

Vascular morphology of the golden hamster spermatic cord.

The morphology of the convoluted testicular artery and the pampiniform plexus of the golden hamster was studied by light microscopy and corrosion cast techniques combined with scanning electron microscopy. The artery was found to be totally enclosed by the pampiniform plexus, except for minor superficial areas where the artery was exposed. Although no direct connection between the artery and the vein was found in the area of apposition, the arterial and venous walls reduced their thickness by sharing a single tunica adventitia, which seemed well suited to the transfer of substances by diffusion. Many band-like structures of the venous walls were found in the deep part of the spermatic cord, suggesting that these may act as barriers to slow down the venous blood velocity. The venous wall here and there showed a stick-like endothelial bridge, suggesting that it may prevent the veins from over distension. In addition to the close relation between the artery and the vein, lymphatic vessels and mast cells were distributed widely within the connective tissue of the arterio-venous walls and venous walls. Mast cells were situated mainly in the area of apposition, especially at the base of the protruding venous wall. These morphological findings suggest that mast cells may be involved in the counter-current transfer mechanism in the spermatic cord of the golden hamster.

The evidence of carotid body in the carotid rete of the shiba goat.

The carotid body was found in/on the cavernous sinus of the carotid rete of the Japanese miniature Shiba goat by light and electron microscopies. The carotid body-like aggregated cell group consisted of two types of cells arranged in the lobule. The first type contained membrane-bound granular vesicles; these cells resembled to the type-I cells in the carotid body. The second type contained no cytoplasmic vesicles, and were located in the periphery, investing the type-I cells. These features appear to be consistent with those of the type-II cells in the carotid body. The nerve ending showed two peculiar features in the intercellular space of type-I cells. The clear vesicle-containing nerve ending showed a structural feature indicative of efferent synapsis. The small granular vesicle-containing nerve ending has been described as a baroreceptor-like ending by VERNA (1979). All of these features are typical characteristics of the carotid body.

Immunohistochemical localization of inhibin and steroidogenic enzymes in the ovary of common tree shrew (Tupaia glis) and northern smooth-tailed tree shrew (Dendrogale murina).

To study the ovarian function of the Order Scandentia, the localization of inhibin and steroidogenic enzymes (3 beta-hydroxysteroid dehydrogenase/isomerase and aromatase) in the ovaries of common tree shrew (Tupaia glis) and northern smooth-tailed tree shrew (Dendrogale murina) was immunohistochemically analysed. As in the results reported for other mammals, inhibin alpha-chain was localized in the follicular epithelium of secondary or Graafian follicles in the two species. The localization of aromatase in the ovary of these two species, however, was different. In the common tree shrew, the aromatase was localized in the thecal cells, whilst in other mammals it is localized in the granulosa cells. These results indicate that in the ovary of the common tree shrew, the oestradiol may be synthesized in the thecal cells.

Production and immunohistochemical characterization of monoclonal antibodies against ovarian antigen of the common tree shrew (Tupaia glis).

To study the ovarian function of the common tree shrew (Tupaia glis), monoclonal antibodies (MAb) against its ovarian antigen were produced. Several positive hybridomas were cloned and MAb-T2C9 (IgG) was examined for reactivity. Strong immunoreactivity of MAb-T2C9 was localized within the cytoplasm of luteal cells and in the theca interna cells of Graafian follicles. MAb-T2C9 reacted also with testicular interstitial cells and adrenocortical cells, except those of the zona glomerulosa. Similar reactions were seen in the goat and sheep. Western blotting analysis of the ovine corpus luteum after reaction with MAb-T2C9 revealed a single positive band of approximately 60 kDa. These findings suggest that MAb-T2C9 recognized protein molecules related to steroid synthesis.

Histochemistry of complex carbohydrate in the major salivary glands of hoary bamboo rats (Rhizomys purinosus).

The major salivary glands (parotid glands, monostomatic sublingual glands and submandibular glands) were obtained from hoary bamboo rats (Rhizomys purinosus) and fixed in Bouin's solution. Paraffin sections were subjected to a battery of staining methods including lectin staining for demonstration of complex carbohydrates. Among the three major salivary glands, unique histochemical features were observed in the submandibular gland. Different from most myomorpha species, submandibular glands of the hoary bamboo rats have two types of secretory cells in the secretory endpieces. One type of cells showed positive reactions with Alcian blue (AB)(pH2.5), periodic acid-Schiff (PAS) and some lectins (peanuts agglutinin, Griffonia simplicifolia I, Maclura pomifera agglutinin). The granular ducts, which exist in animals belonging to suborder myomorpha, were not observed in the submandibular glands of this animal.

Cardiac musculature of the cranial vena cava in the common tree shrew (Tupaia glis).

Cardiac musculature of the cranial vena cava in the common tree shrew (Tupaia glis) was examined by light and transmission electron microscopy. The common tree shrew has well developed cardiac myocyte layers in the tunica media of the cranial vena cava, extending from the right atrium to the root of the subclavian vein. Because the common tree shrew belongs to a primitive group of mammals, the occurrence of cardiac musculature in the cranial vena cava may be a common feature in lower mammals. The development of this musculature indicates that active contraction of the cranial vena cava wall occurs in this species. Electron micrographs showed the typical ultrastructure of myocytes and nerve endings. These observations suggest that this musculature may serve as a regulatory pump for the return of venous blood to the right atrium and as a blood reservoir system under conditions of rapid heart rate. Additionally, the presence of atrial natriuretic polypeptide (ANP) was also demonstrated in the myocytes of the vena cava immunohistochemically. These findings show that the cardiac endocrine organ for ANP develops even in the principal veins including the cranial vena cava.

An ultrastructural and lectin-histochemical study on the seminiferous epithelium of the common tree shrew (Tupaia glis).

The seminiferous epithelium of the common tree shrew (Tupaia glis) was investigated using transmission electron microscopy and lectin-histochemistry. It was compared with that of shrews examined in previous studies. Some peculiar structures were detected in the tree shrew spermatid at the electron microscopic level. The most characteristic feature was the disposition of mitochondria in early spermatids. In cap and early acrosome-phase spermatids, mitochondria accumulated in one area of the spermatid cytoplasm and then dispersed in the late acrosome phase. Subsequently they again clustered to form the middle piece of spermatozoa. While the lamellar structure was clearly seen in the caudal region of the spermatid nucleus, it gradually disappeared during the process of elongation. The dilated area in the postacrosomal space of early round spermatids was also characteristic. The dilation was not detected in elongated spermatids. These structures were not recognised in the seminiferous epithelium of shrews. With respect to lectin histochemistry, the binding patterns in the spermatid acrosome of the tree shrew were similar to those of the musk shrew. However, PNA and BPA, which reacted with the Sertoli cell cytoplasm of the musk shrew, showed no reaction in the tree shrew Sertoli cell. Thus, except for some lectin binding patterns in the spermatid acrosome, no close similarity was recognised in the morphology of the seminiferous epithelium between tree shrews and shrews.

Cardiac-like musculature of the intrapulmonary venous wall of the long-clawed shrew (Sorex unguiculatus), common tree shrew (Tupaia glis) and common marmoset (Callithrix jacchus).

BACKGROUND: The cardiac-like musculature is distributed not only in the heart wall but also in the intrapulmonary venous wall in a few species of insectivores. It has been suggested that the evolutionary origin of venous cardiac-like musculature may be traceable to a basic stock of certain mammalian lines of descent. So, it is important to clarify whether the musculature may be a common structure in lower mammals from insectivores to primates and to examine the functional significance of the structure. METHODS: The distribution of cardiac-like musculature in the intrapulmonary venous wall of the long-clawed shrew (Sorex unguiculatus), common tree shrew (Tupaia glis), and common marmoset (Callithrix jacchus) was observed by light and electron microscopy. The presence of atrial natriuretic polypeptide (ANP) was examined in the musculature by immunohistochemistry. RESULTS: All three species contained cardiac-like myocytes in the tunica media of intrapulmonary venous wall. In the common tree shrew and the common marmoset, cardiac-like musculature was found in the small intrapulmonary vein of 150-200 microns in diameter, while, in the long-clawed shrew, it was distributed even in the intrapulmonary vein of 30 microns in diameter. Ultrastructure of myocytes was fundamentally similar to that of atrial myocytes in the long-clawed shrew and the common tree shrew. The presence of atrial natriuretic polypeptide (ANP) was also demonstrated by immunohistochemistry in the intrapulmonary venous walls of common tree shrews. CONCLUSIONS: The results indicate that the pulmonary venous cardiac-like musculature occurs in the lower eutherian mammals from insectivores to primitive primates. The musculature is thought to act as a regulatory pump in pulmonary circulation and as an endocrine apparatus of ANP.