Results of hyperamplification of centrosomes in naturally developing tumors of dogs.

OBJECTIVE: To evaluate results of centrosome hyperamplification in naturally developing tumors of dogs. SAMPLE POPULATION: Tumor specimens from 9 dogs with tumors (rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myxosarcoma, and mammary gland tumor) and 2 canine osteosarcoma cell lines. PROCEDURE: 3 antibodies for centrosome proteins (ie, anti-gamma-tubulin, anti-BRCA1, and anti-pericentrin) were used for immunohistochemical analysis. Double immunostaining for centrosomes was used to confirm the specificity of these antibodies for centrosomes. Mutational analysis of the canine p53 gene was carried out by polymerase chain reaction-single-strand conformation polymorphism analysis, and expression of canine MDM2 protein was evaluated by use of immunohistochemical analysis, using anti-MDM2 antibody. RESULTS: Immunohistochemical analysis of dog osteosarcoma cell lines with apparent aneuploidy revealed frequent hyperamplification of centrosomes in the osteosarcoma cell lines. Similar hyperamplified centrosomes were detected in the tumor tissues from all of the 9 tumors. The frequency of cells with hyperamplified centrosomes (3 to 20/cell) in each tumor tissue ranged from 9.50 to 48.1%, whereas centrosome hyperamplification was not observed in normal lymph nodes from these dogs. In 8 of the 9 tumors, mutation of p53 gene or overexpression of MDM2, or both, was detected. CONCLUSIONS AND CLINICAL RELEVANCE: Various types of naturally developing tumors in dogs often have hyperamplification of centrosomes associated with chromosome instability. Hyperamplification of centrosomes is a novel tumor marker for use in cytologic and histologic examinations of clinical specimens obtained from dogs.     

The Arterial Supply of the Male Reproductive System in the Hamster

The major arteries, the testicular, deferential, prostatic, internal pudendal and external pudendal arteries, supply the male genital organs of the hamster. The same arterial patterns are found in three strains, except that the external pudendal artery arises from the external iliac artery or internal iliac artery in the APG strain, but only from the external iliac artery in the CBN and ACN strains. The deferential and prostatic arteries form the arterial loop on the dorsal surface of the ventrolateral lobe of the prostate. This loop may play some importance parts in the functional interaction between the epididymis and the prostate. The caudal epididymal artery arises more frequently from the spermatic cordai portion of the testicular artery than from the cranial epididymal artery, and it never arises directly from the abdominal part of the testicular artery as reported in the rat, mouse and rabbit. The deferential artery may join with the cranial vesicular artery to form the common trunk (umbilical artery), but sometimes these arteries arise directly from the internal iliac artery.     

Analysis of various antigens in golden hamster testis by monoclonal antibodies.

A total of 38 hybridomas producing monoclonal antibodies (mAbs) was established by immunizing BALB/c mice with extracts of the golden hamster testis. Six mAbs stained the acrosome of developing spermatids by immunofluorescence. Two mAbs (1A11 and 4D8) reacted with spermatid components other than acrosome. The mAbs 1C9 and 4D3 recognized a 103 kilodalton (kDa) protein on immunoblots, and were reactive to spermatocytes and early spermatids, but not to late spermatids and spermatozoa. This finding suggests that the protein functions for meiosis or early spermiogenesis. Four mAbs (3G2, 2E5, 2G3, and 3F10) stained all stages of spermatogenic cells. The remaining 24 mAbs showed a positive reaction to the basement membrane of the seminiferous tubule. Two of them, 3D6 and 3E5, recognized approximately 150 kDa major proteins, indicating that the antigen is an extracellular matrix.     

Isolation of a human erythrocyte-adapted substrain of Babesia rodhaini and analysis of the merozoite surface protein gene sequences

Babesia rodhaini is a rodent hemoparasite closely related to B. microti, the major causative agent of human babesiosis. We tested the infectivity of B. rodhaini for human erythrocytes by using the SCID mouse model in which the circulating erythrocytes were replaced with those of humans. Initially, parasites grew very poorly in the mouse model, but a variant capable of propagating in human erythrocytes emerged after an adaptation period of three weeks. In an attempt to identify parasite proteins involved in the alteration of host cell preference, an expression cDNA library of B. rodhaini was constructed and screened with immune mouse sera. Although we were able to obtain three merozoite surface protein (MSP) genes, sequences of these genes from both the parental strain and human erythrocyte-adapted substrain were identical. Our results suggest that B. rodhaini has potential ability to infect human erythrocytes, but development of this ability may not be brought about by an amino acid change in MSPs.     

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.