Patterns of sequence variation in the mitochondrial D-loop region of shrews

Direct sequencing of the mitochondrial displacement loop (D-loop) of shrews (genus Sorex) for the region between the tRNA(Pro) and the conserved sequence block-F revealed variable numbers of 79-bp tandem repeats. These repeats were found in all 19 individuals sequenced, representing three subspecies and one closely related species of the masked shrew group (Sorex cinereus cinereus, S. c. miscix, S. c. acadicus, and S. haydeni) and an outgroup, the pygmy shrew (S. hoyi). Each specimen also possessed an adjacent 76-bp imperfect copy of the tandem repeats. One individual was heteroplasmic for length variants consisting of five and seven copies of the 79-bp tandem repeat. The sequence of the repeats is conducive to the formation of secondary structure. A termination-associated sequence is present in each of the repeats and in a unique sequence region 5' to the tandem array as well. Mean genetic distance between the masked shrew taxa and the pygmy shrew was calculated separately for the unique sequence region, one of the tandem repeats, the imperfect repeat, and these three regions combined. The unique sequence region evolved more rapidly than the tandem repeats or the imperfect repeat. The small genetic distance between pairs of tandem repeats within an individual is consistent with a model of concerted evolution. Repeats are apparently duplicated and lost at a high rate, which tends to homogenize the tandem array. The rate of D- loop sequence divergence between the masked and pygmy shrews is estimated to be 15%-20%/Myr, the highest rate observed in D-loops of mammals. Rapid sequence evolution in shrews may be due either to their high metabolic rate and short generation time or to the presence of variable numbers of tandem repeats.      

大鼠胼胝体内神经肽Y免疫反应阳性纤维的发育

本实验用免疫组织化学ＡＢＣ法研究了大鼠胼胝体内神经肽Ｙ免疫反应阳性（ＮＰＹ－ＩＲ）纤维的生后发育。结果发现,许多ＮＰＹ－ＩＲ纤维在大鼠出生时便存在于胼胝体内。ＮＰＹ－ＩＲ胼胝体纤维的密度在生后１周内继续逐渐增高,在第２周内达到最高峰。之后,ＮＰＹ－ＩＲ胼胝体纤维的密度逐渐下降,至第３周末时接近成年时的水平,即仅有少量ＮＰＹ－ＩＲ纤维存在于胼胝体内。这些结果提示在大鼠早期生后发育过程中许多ＮＰＹ－ＩＲ胼胝体纤维是暂时性的,其作用可能与大脑皮质的机能发育有关。     

Tissue proteomics of the low‐molecular weight proteome using an integrated cLC‐ESI‐QTOFMS approach

Analysis of the protein/peptide composition of tissue has provided meaningful insights into tissue biology and even disease mechanisms. However, little has been published regarding top down methods to investigate lower molecular weight (MW) (500–5000 Da) species in tissue. Here, we evaluate a tissue proteomics approach involving tissue homogenization followed by depletion of large proteins and then cLC‐MS (where c stands for capillary) analysis to interrogate the low MW/low abundance tissue proteome. In the development of this method, sheep heart, lung, liver, kidney, and spleen were surveyed to test our ability to observe tissue differences. After categorical tissue differences were demonstrated, a detailed study of this method's reproducibility was undertaken to determine whether or not it is suitable for analyzing more subtle differences in the abundance of small proteins and peptides. Our results suggest that this method should be useful in exploring the low MW proteome of tissues.     

An association between the acute phase response and patterns of antigen induced T cell proliferation in juvenile idiopathic arthritis

The aim of this research was to determine whether all memory T cells have the same propensity to migrate to the joint in patients with juvenile idiopathic arthritis. Paired synovial fluid and peripheral blood mononuclear cell proliferative responses to a panel of antigens were measured and the results correlated with a detailed set of laboratory and clinical data from 39 patients with juvenile idiopathic arthritis. Two distinct patterns of proliferative response were found in the majority of patients: a diverse pattern, in which synovial fluid responses were greater than peripheral blood responses for all antigens tested; and a restricted pattern, in which peripheral blood responses to some antigens were more vigorous than those in the synovial fluid compartment. The diverse pattern was generally found in patients with a high acute phase response, whereas patients without elevated acute phase proteins were more likely to demonstrate a restricted pattern. We propose that an association between the synovial fluid T cell repertoire and the acute phase response suggests that proinflammatory cytokines may influence recruitment of memory T cells to an inflammatory site, independent of their antigen specificity. Additionally, increased responses to enteric bacteria and the presence of αEβ7 T cells in synovial fluid may reflect accumulation of gut associated T cells in the synovial compartment, even in the absence of an elevated acute phase response. This is the first report of an association between the acute phase response and the T cell population recruited to an inflammatory site.     

Artificial selection of rams for sexual performance and its effect on the sexual behavior and fecundity of male and female progeny

The objective of this study was to determine the effect of a single generation of artificial selection for sexual performance in rams on the sexual behavior and fecundity of their male and female progeny. Ninety-two ram lambs born to sires selected for either high or low sexual performance were evaluated for their sexual behaviors at approximately 8 months of age when individually exposed to four estrous ewes for 30min in four weekly serving capacity tests. Number of mounts and successful matings (ejaculations) were recorded. Fourteen of the 17 high-performing ram lambs identified were sired by high-performing sires, whereas 22 of 37 low-performing ram lambs were sired by low-performing sires (P<0.01). Sons of high-performing sires exhibited more ejaculations (P<0.04) and more mounts without ejaculation (P<0.02) than sons of low-performing sires. The two groups of ram lambs did not differ in mating efficiency (ratio of ejaculations to total mounts). Daughters of high-performing rams (N=79) exhibited their first behavioral estrus approximately 8 days earlier than daughters (N=61) of low-performing sires (P<0.005). Ovulation rates for the two groups of ewe lambs did not differ (P=0.55). It was concluded that there was sufficient genetic variation in the population of sheep studied to obtain a significant response to selection for ram sexual performance in both male and female offspring in a single generation.     

Zinc is sufficiently abundant within mammalian sperm nuclei to bind stoichiometrically with protamine 2

Although studies have demonstrated that zinc can bind to sperm nuclear proteins, specifically protamine 2, it has not been shown that the metal is sufficiently abundant inside the sperm nucleus to interact stoichiometrically with these proteins. In this study proton-induced X-ray emission (PIXE) has been used to measure the amount of sulfur and zinc within the nuclei of individual sperm cells to infer the stoichiometry of zinc binding to protamine 2 in six species of mammal: bull, chinchilla, stallion, hamster, human, and mouse (protamine 2 comprises from 0% (bull) to 67% (mouse) of the protamine present in the sperm of these animals). Using the sulfur mass and electrophoretic data on the relative proportion of protamine 1 and protamine 2 in the sperm chromatin of these species, the protamine 1, protamine 2, and total protamine contents within each species sperm nuclei have been determined. The PIXE measurements reveal that the zinc content of the sperm nucleus varies proportionately with the protamine 2 content of sperm chromatin. PIXE analyses of hamster protamines extracted under conditions that appear to at least partially preserve zinc binding also confirm that the majority of the metal is bound to protamine. In five of the species examined, sufficient zinc is present for each protamine 2 molecule to bind one zinc. The results obtained for chinchilla sperm, conversely, indicate the chinchilla protamine 2 molecule may interact differently with zinc. Chinchilla sperm only contain enough zinc for one atom to be bound to two protamine 2 molecules.     

Sexual performance of twin ram lambs and the effect of number and sex of contemporary siblings

The objectives of this investigation were to determine the degree of correspondence in the sexual performance of twin male sheep and to compare the sexual performance of ram lambs born co-twin to another male (M-M), co-twin to a female (M-F) and as single (S) offspring. Individual ram lambs (N=117), including 13 M-M twin sets, were exposed to four estrous ewes for 30 min at 7-day intervals over a 4-week period at 8 to 9 months of age.Frequencies of ejaculation (serving capacity) and mounts without ejaculation were recorded. M-M males averaged 2.9 ejaculations and 13.1 mounts per test. Individuals within 9 of the 13 M-M twin sets (69%) differed by an average of 0.5 or fewer ejaculations per test. Although variability within M-M twin sets was very low, variability between the 13 M-M twin sets was also low (coefficient of variation=10.6%). Consequently, frequencies of both ejaculation and mounting (without ejaculation) within the 13 twin sets were not correlated (P=0.50 and P=0.10, respectively). When the progeny of individual sires were compared, mounting frequencies were correlated (P<0.02) but not ejaculation rates (P=0.72). M-M, M-F and S ram lambs did not differ for either ejaculation frequency (P=0.26) or mounts without ejaculation (P=0.98). It was concluded that the sexual performance of related individuals is likely to be very similar and that the number and sex of contemporary siblings does not necessarily influence rates of mounting and ejaculation.